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Academic OrganizationAcademic Assembly School of Medicine and Health Sciences Institute of Health ScienceTEL
Education and Research OrganizationSchool of Medicine Department of Biomedical Laboratory SciencesFAX
PositionProfessorMail Address
Address3-1-1, Asahi, Matsumoto, Nagano, JAPANWeb site



Research Field
Hematology and oncology
Keywords:pure red cell aplasia , large grnanular lymphocyte , bone marrow failure , NK cell lymphoma/leukemia
Current Subject
Keywords:pathophysiology of NK cell lymphoma/leukemia
molecular pathogenesis of bone marrow failure including pure red cell aplasia
Keywords:bone marrow failure , pure red cellaplasia , T cell
Academic Societies
Academic Societies
American Society of Hematology
American College of Physicians
Japanese Society of Hematology
Academic Background
Graduate School
Shinshu University , (Graduate School, Division of Medicine) , 1994

Shinshu University , (Faculty of Medicine) , 1985

Doctor of Medicine , 信州大学
1996 , 松医会賞受賞


Books, Articles, etc.
血小板 GPIb b遺伝子異常とBernard-Soulier症候群・大血小板
中外医学社,東京 2003

The hepatic niche leads to aggressive natural killer cell leukemia proliferation through the transferrin-transferrin receptor 1 axis.
Blood,142(4):352-364 2023(Jul. 27)
Author:Kazuaki Kameda; Ryo Yanagiya; Yuji Miyatake; Joaquim Carreras; Hiroshi Higuchi; Hiromichi Murayama; Takashi Ishida; Asahi Ito; Shinsuke Iida; Noriko Fukuhara; Hideo Harigae; Yuki Fujioka; Naoto Takahashi; Hidenori Wada; Fumihiro Ishida; Hideyuki Nakazawa; Rei Ishihara; Yuki Murakami; Hiroyuki Tagawa; Tadashi Matsuura; So Nakagawa; Sadahiro Iwabuchi; Shinichi Hashimoto; Ken-Ichi Imadome; Naoya Nakamura; Kenichi Ishizawa; Yoshinobu Kanda; Kiyoshi Ando; Ai Kotani
Abstract:Aggressive natural killer cell leukemia (ANKL) is a rare lymphoid neoplasm frequently associated with Epstein-Barr virus, with a disastrously poor prognosis. Owing to the lack of samples from patients with ANKL and relevant murine models, comprehensive investigation of its pathogenesis including the tumor microenvironment (TME) has been hindered. Here we established 3 xenograft mice derived from patients with ANKL (PDXs), which enabled extensive analysis of tumor cells and their TME. ANKL cells primarily engrafted and proliferated in the hepatic sinusoid. Hepatic ANKL cells were characterized by an enriched Myc-pathway and proliferated faster than those in other organs. Interactome analyses and in vivo CRISPR-Cas9 analyses revealed transferrin (Tf)-transferrin receptor 1 (TfR1) axis as a potential molecular interaction between the liver and ANKL. ANKL cells were rather vulnerable to iron deprivation. PPMX-T003, a humanized anti-TfR1 monoclonal antibody, showed remarkable therapeutic efficacy in a preclinical setting using ANKL-PDXs. These findings indicate that the liver, a noncanonical hematopoietic organ in adults, serves as a principal niche for ANKL and the inhibition of the Tf-TfR1 axis is a promising therapeutic strategy for ANKL.

Tγδ LGLL identifies a subset with more symptomatic disease: analysis of an international cohort of 137 patients.
Blood,141(9):1036-1046 2023(Mar. 02)
Author:Gregorio Barilà; Angela Grassi; HeeJin Cheon; Antonella Teramo; Giulia Calabretto; Jasmanet Chahal; Cristina Vicenzetto; Julia Almeida; Bryna C Shemo; Min Shi; Vanessa Rebecca Gasparini; Noemi Munoz-Garcia; Cédric Pastoret; Hideyuki Nakazawa; Kazuo Oshimi; Lubomir Sokol; Fumihiro Ishida; Thierry Lamy; Alberto Orfao; William G Morice; Thomas P Loughran; Gianpietro Semenzato; Renato Zambello
Abstract:Tγδ large granular lymphocyte leukemia (LGLL) is a rare variant of T-cell LGLL (T-LGLL) that has been less investigated as compared with the more frequent Tαβ LGLL, particularly in terms of frequency of STAT3 and STAT5b mutations. In this study, we characterized the clinical and biological features of 137 patients affected by Tγδ LGLL; data were retrospectively collected from 1997 to 2020 at 8 referral centers. Neutropenia and anemia were the most relevant clinical features, being present in 54.2% and 49.6% of cases, respectively, including severe neutropenia and anemia in ∼20% of cases each. Among the various treatments, cyclosporine A was shown to provide the best response rates. DNA samples of 97 and 94 cases were available for STAT3 and STAT5b mutation analysis, with 38.1% and 4.2% of cases being mutated, respectively. Clinical and biological features of our series of Tγδ cases were also compared with a recently published Tαβ cohort including 129 cases. Though no differences in STAT3 and STAT5b mutational frequency were found, Tγδ cases more frequently presented with neutropenia (P = .0161), anemia (P < .0001), severe anemia (P = .0065), and thrombocytopenia (P = .0187). Moreover, Vδ2- cases displayed higher frequency of symptomatic disease. Overall, Tγδ cases displayed reduced survival with respect to Tαβ cases (P = .0017). Although there was no difference in STAT3 mutation frequency, our results showed that Tγδ LGLL represents a subset of T-LGLL characterized by more frequent symptoms and reduced survival as compared with Tαβ LGLL.

Immature Platelet Fraction and Its Kinetics in Neonates.
Journal of pediatric hematology/oncology,45(2):e249-e253 2023(Mar. 01)
Author:Jun Kobayashi; Yuka Takezawa; Shoji Saito; Noriko Kubota; Kazuo Sakashita; Yozo Nakazawa; Yumiko Higuchi; Minoru Tozuka; Fumihiro Ishida
Abstract:Thrombocytopenia is a common abnormality encountered in the neonatal period, and immature platelet fraction (IPF) may be an informative indicator of thrombopoiesis; however, data on IPF in neonates are scarce. To define reference intervals (RIs) and factors affecting IPF in neonates, we measured the IPF of 533 consecutive neonates. With a multiple regression analysis of 330 newborns with normal platelet counts at birth, premature delivery, neonatal asphyxia, intrauterine infection, chromosomal abnormalities, and respiratory disorders were identified as independent factors for IPF%. The RIs of IPF% and absolute IPF value in neonates were determined to be 1.3% to 5.7% and 3.2 to 14.5×10 9 /L, respectively. On day 14 after birth, IPF% increased to twice the value at birth and thereafter returned to the previous value on day 28. Reticulocyte counts, in contrast, were the lowest at day 14. IPF% was increased in 16 thrombocytopenic patients with various clinical conditions, especially those with immune-mediated thrombocytopenia. IPF in neonates may be evaluated essentially based on the same RIs as in adults, although some precautions must be taken when evaluating IPF in neonates in the first 2 weeks of life. IPF may be useful for evaluating thrombopoiesis and thrombocytopenia in neonates.

A Diagnostic Impact of Serum Autotaxin Levels in Patients with Bone Marrow Fibrosis.
Clinical lymphoma, myeloma & leukemia,23(2):e117-e124 2023(Feb.)
Author:Hideyuki Nakazawa; Hiroko Kaiume; Koji Igarashi; Tomoo Yamazaki; Takeji Umemura; Naoko Asano; Takeshi Uehara; Fumihiro Ishida
Abstract:BACKGROUND: Bone marrow (BM) fibrosis is a condition characterized by deposition of reticulin and collagen fibers in BM. It may confer a poor prognosis in some of hematological malignancies. However, the relationship between fibrosis and the disease pathology is not fully understood and no biomarkers for BM fibrosis are available in clinical practice. Autotaxin (ATX) is a secreted enzyme that is associated with various pathophysiological responses, including fibrosis. We conducted a pilot study to investigate the serum ATX levels in various hematological disorders in patients with or without BM fibrosis. PATIENTS AND METHODS: The serum levels of ATX in a total of 198 patients with hematological disorders and 160 healthy subjects were analyzed. Because of sexual difference in ATX level, the ATX ratio-determined by dividing the ATX level by the mean value of ATX of control subjects of the same sex-was calculated for further comparative analysis. A trephine biopsy samples from 53 patients were also evaluated to determine the Reticulin Fibrosis Index and Collagen Fibrosis Index of each sample. RESULTS: In comparison to the control group, the ATX ratio was significantly higher in patients, especially those with malignant lymphoma. The ATX ratio in lymphoma patients with BM fibrosis was significantly higher than that in patients without BM fibrosis. The Collagen Fibrosis Index showed statistically significant negative correlation with the ATX ratio. CONCLUSION: Our results suggest that the ATX ratio may be a candidate diagnostic biomarker for BM fibrosis in selected patients, including those with malignant lymphoma.

Incidence of acquired pure red cell aplasia: a nationwide epidemiologic analysis with 2 registry databases in Japan.
Blood advances,6(24):6282-6290 2022(Dec. 27)
Author:Hideyuki Nakazawa; Kaoko Sakai; Akiko Ohta; Naohito Fujishima; Akira Matsuda; Kohei Hosokawa; Fumi Nakamura; Shinji Nakao; Kinuko Mitani; Fumihiro Ishida
Abstract:Acquired pure red cell aplasia (PRCA) is a rare syndrome characterized by anemia with reticulocytopenia and a marked reduction in erythroid precursors. Given its rarity, the true incidence is largely unknown, and epidemiological data representing the general population, with a description of the full spectrum of etiologies, are scarce. An epidemiological study on PRCA in Japan conducted 30 years ago estimated the annual incidence as 0.3 per million. To update the data and investigate the incidence and demographics of PRCA, we conducted a nationwide epidemiological study using the Japanese Society of Hematology (JSH) Hematologic Disease Registry, a hematologic disease registration database managed by the JSH and the Diagnosis Procedure Combination (DPC) study data available at a website of the Ministry of Health, Labor, and Welfare (MHLW) of Japan. A total of 1055 patients with newly diagnosed acquired PRCA were identified between 2012 and 2019, and the average annual incidence was calculated at 1.06 (95% confidence interval [CI], 0.83-1.28) per million. The median age was 73 (range, 18-99) years. The female-to-male ratio was 1.5:1, and the female predominance was most prominent in the child-bearing age group. Sixty-nine percent of acquired PRCA was idiopathic. The incidence of PRCA was approximately 20% of that of aplastic anemia (AA) during the same period. Approximately 0.98 patients per million per year (95% CI, 0.89-1.07) required hospitalization for the treatment of PRCA. These results are expected to contribute to the discussion of resource allocation for PRCA in the aging population in many countries, including Japan.

Comparative analysis of humoral responses to BNT162b2 vaccine among patients with hematologic disorders and organ transplant recipients.
Transplant immunology,75:101713-101713 2022(Dec.)
Author:Hideyuki Nakazawa; Kaoko Sakai; Yuriko Sudo; Ryohei Iwabuchi; Hitoshi Sakai; Sayaka Nishina; Toru Kawakami; Fumihiro Kawakami; Shuji Matsuzawa; Toshiro Ito; Mari Kitahara; Yuji Kamijo; Takeji Umemura; Atsuhito Ushiki; Shinichiro Kanai; Hiroyuki Tsuchiya; Fumihiro Ishida
Abstract:Vaccination against SARS-COV-2 is considered the most promising approach to curbing the pandemic. Patients with an immunocompromised state, such as those with hematological malignancies and organ transplantation recipients, are considered more susceptible to infection, but these at-risk patients were underrepresented in early clinical trials for vaccination. Although a growing body of studies suggests that the humoral response to COVID-19 vaccination in each of these at-risk groups of patients may be suboptimal in comparison to healthy controls, a clinical and strategic information for the further comparative analysis among these groups is not fully described. The humoral responses after two doses of BNT162b2 vaccination were evaluated in a total of 187 patients either with allogeneic hematopoietic transplantation, with renal transplantation, with anti-CD20 antibody therapy, or with anti-CD38 antibody therapy, and in 66 healthy controls. The early response at one to three months after vaccination was significantly inferior among patients with renal transplantation, patients with anti-CD20 antibody therapy, and patients with anti-CD38 antibody therapy in comparison to healthy control. But the patients with allogeneic hematopoietic transplantation showed early humoral response comparable to healthy control. The late response at 6 months after vaccination was still suboptimal among patients with renal transplantation and patients with anti-CD20 therapy. Among our patient group, renal transplant recipients had the lowest antibody titers after vaccination regardless of timing of vaccination. Patients who had received allogeneic hematopoietic transplantation attained a comparable serological response to the control group especially if they are vaccinated >300 days after transplantation, but the response was suboptimal if the vaccination was within 300 days after transplantation. Our results may provide policy makers with critical information for the further stratification of at-risk groups, helping contribute to a better allocation of resources, including additional booster vaccination.

Anti-COX-2 autoantibody is a novel biomarker of immune aplastic anemia
LEUKEMIA,36(9):2317-2327 2022(Sep.)
Author:Kelkka, Tiina; Tyster, Mikko; Lundgren, Sofie; Feng, Xingmin; Kerr, Cassandra; Hosokawa, Kohei; Huuhtanen, Jani; Keranen, Mikko; Patel, Bhavisha; Kawakami, Toru; Maeda, Yuka; Nieminen, Otso; Kasanen, Tiina; Aronen, Pasi; Yadav, Bhagwan; Rajala, Hanna; Nakazawa, Hideyuki; Jaatinen, Taina; Hellstrom-Lindberg, Eva; Ogawa, Seishi; Ishida, Fumihiro; Nishikawa, Hiroyoshi; Nakao, Shinji; Maciejewski, Jaroslaw; Young, Neal S.; Mustjoki, Satu
Abstract:In immune aplastic anemia (IAA), severe pancytopenia results from the immune-mediated destruction of hematopoietic stem cells. Several autoantibodies have been reported, but no clinically applicable autoantibody tests are available for IAA. We screened autoantibodies using a microarray containing >9000 proteins and validated the findings in a large international cohort of IAA patients (n = 405) and controls (n = 815). We identified a novel autoantibody that binds to the C-terminal end of cyclooxygenase 2 (COX-2, aCOX-2 Ab). In total, 37% of all adult IAA patients tested positive for aCOX-2 Ab, while only 1.7% of the controls were aCOX-2 Ab positive. Sporadic non-IAA aCOX-2 Ab positive cases were observed among patients with related bone marrow failure diseases, multiple sclerosis, and type I diabetes, whereas no aCOX-2 Ab seropositivity was detected in the healthy controls, in patients with non-autoinflammatory diseases or rheumatoid arthritis. In IAA, anti-COX-2 Ab positivity correlated with age and the HLA-DRB1*15:01 genotype. 83% of the >40 years old IAA patients with HLA-DRB1*15:01 were anti-COX-2 Ab positive, indicating an excellent sensitivity in this group. aCOX-2 Ab positive IAA patients also presented lower platelet counts. Our results suggest that aCOX-2 Ab defines a distinct subgroup of IAA and may serve as a valuable disease biomarker.

Somatic mutations in acquired pure red cell aplasia.
Seminars in hematology,59(3):131-136 2022(Jul.)
Author:Toru Kawakami; Hideyuki Nakazawa; Fumihiro Ishida
Abstract:Acquired pure red cell aplasia (PRCA) is a syndrome characterized by anemia and a marked reduction of erythroid progenitor cells with various etiologies. The 3 major subtypes of PRCA are idiopathic PRCA, large granular lymphocytic leukemia-associated PRCA and thymoma-associated PRCA, which are thought to be caused by a T-cell-mediated mechanism. In these 3 subtypes, an expansion of clonal cytotoxic T cells is often detected. In addition, those T cells recurrently harbor somatic mutations of STAT3, a gene coding one of the important signal transducers in the JAK/STAT system. Somatic mutations of clonal hematopoiesis (CH)-related genes, including epigenetic modifying genes, have also been reported, however, the data are still not mature enough upon which to draw conclusion, Somatic mutations of STAT3 and CH-related genes may be unique characteristics of acquired PRCA. However, their involvement in dyserythropoiesis or clinical relevance to the clinical course of those somatic mutations. Mutational landscapes, their involvements in dyserythropoiesis and clinical relevance in acquired PRCA remains unclear, and further investigation is needed.

T cell clonal expansion and STAT3 mutations: a characteristic feature of acquired chronic T cell-mediated pure red cell aplasia
Author:Kawakami, Fumihiro; Kawakami, Toru; Yamane, Taku; Maruyama, Masae; Kobayashi, Jun; Nishina, Sayaka; Sakai, Hitoshi; Higuchi, Yumiko; Hamanaka, Kazutoshi; Hirokawa, Makoto; Nakao, Shinji; Nakazawa, Hideyuki; Ishida, Fumihiro
Abstract:Acquired chronic pure red cell aplasia (PRCA) develops idiopathically or in association with other medical conditions, including T cell large granular lymphocytic leukemia (T-LGLL) and thymoma. T cell dysregulation is considered a cardinal pathogenesis of PRCA, but genetic-phenotypic associations in T cell abnormalities are largely unclear. We evaluated an extended cohort of 90 patients with acquired PRCA, including 26 with idiopathic, 36 with T-LGLL-associated and 15 with thymoma-associated PRCA, for their T cell immuno-phenotypes, clonalities and STAT3 mutations. TCR repertoire skewing of CD8+ T cells was detected in 37.5% of idiopathic, 66.7% of T-LGLL-associated and 25% of thymoma-associated PRCA patients, and restriction to Vβ1 was most prominent (41%). Clonalities of TCRβ or γ chain and STAT3 mutational status were statistically associated (P = 0.0398), and they were detected in all three subtypes. The overall response rate to cyclosporin A was 73.9%, without significant difference by subtypes nor STAT3 mutational status. The T cell dysregulations, such as TCR repertoire skewing with predominant Vβ1 usage, clonality and STAT3 mutations, were frequently found across the subtypes, and the close associations between them suggest that these T cell derangements reflect a common pathophysiological mechanism among these PRCA subtypes.

Identification of novel STAT5B mutations and characterization of TCRβ signatures in CD4+ T-cell large granular lymphocyte leukemia
Blood Cancer Journal,12(2):31-31 2022(Feb.)
Author:Dipabarna Bhattacharya; Antonella Teramo; Vanessa Rebecca Gasparini; Jani Huuhtanen; Daehong Kim; Jason Theodoropoulos; Gianluca Schiavoni; Gregorio Barilà; Cristina Vicenzetto; Giulia Calabretto; Monica Facco; Toru Kawakami; Hideyuki Nakazawa; Brunangelo Falini; Enrico Tiacci; Fumihiro Ishida; Gianpietro Semenzato; Tiina Kelkka; Renato Zambello; Satu Mustjoki
Abstract:AbstractCD4+ T-cell large granular lymphocyte leukemia (T-LGLL) is a rare subtype of T-LGLL with unknown etiology. In this study, we molecularly characterized a cohort of patients (n = 35) by studying their T-cell receptor (TCR) repertoire and the presence of somatic STAT5B mutations. In addition to the previously described gain-of-function mutations (N642H, Y665F, Q706L, S715F), we discovered six novel STAT5B mutations (Q220H, E433K, T628S, P658R, P702A, and V712E). Multiple STAT5B mutations were present in 22% (5/23) of STAT5B mutated CD4+ T-LGLL cases, either coexisting in one clone or in distinct clones. Patients with STAT5B mutations had increased lymphocyte and LGL counts when compared to STAT5B wild-type patients. TCRβ sequencing showed that, in addition to large LGL expansions, non-leukemic T cell repertoires were more clonal in CD4+ T-LGLL compared to healthy. Interestingly, 25% (15/59) of CD4+ T-LGLL clonotypes were found, albeit in much lower frequencies, in the non-leukemic CD4+ T cell repertoires of the CD4+ T-LGLL patients. Additionally, we further confirmed the previously reported clonal dominance of TRBV6-expressing clones in CD4+ T-LGLL. In conclusion, CD4+ T-LGLL patients have a typical TCR and mutation profile suggestive of aberrant antigen response underlying the disease.

Single-cell characterization of leukemic and non-leukemic immune repertoires in CD8(+) T-cell large granular lymphocytic leukemia
Author:Huuhtanen, Jani; Bhattacharya, Dipabarna; Lonnberg, Tapio; Kankainen, Matti; Kerr, Cassandra; Theodoropoulos, Jason; Rajala, Hanna; Gurnari, Carmelo; Kasanen, Tiina; Braun, Till; Teramo, Antonella; Zambello, Renato; Herling, Marco; Ishida, Fumihiro; Kawakami, Toru; Salmi, Marko; Loughran, Thomas; Maciejewski, Jaroslaw P.; Lahdesmaki, Harri; Kelkka, Tiina; Mustjoki, Satu

[Clinical and pathophysiological features of acquired pure red cell aplasia: based on the concept of T-cell dysregulations].
[Rinsho ketsueki] The Japanese journal of clinical hematology,63(8):893-898 2022
Author:Fumihiro Ishida
Abstract:Acquired pure red cell aplasia (PRCA) develops in a variety of contexts and thus, should be regarded as a syndrome. The three major subtypes of acquired PRCA are idiopathic PRCA, T cell large granular lymphocytic leukemia (T-LGLL)-associated PRCA, and thymoma-associated PRCA. Although the Japanese National Research Group on Idiopathic Bone Marrow Failure Syndromes of the Ministry of Health, Labor and Welfare of Japan has made significant contributions to our understanding of PRCA, details of its clinical characteristics, and pathophysiological mechanisms remain largely unknown. A recent epidemiological analysis using the JSH Hematologic Disease Registry revealed that approximately 100 new cases with acquired PRCA were diagnosed annually in Japan, which was higher than previously thought to be. The median age of the patients was 73 years. A prospective observational study on chronic PRCA (PRCA2016) is currently ongoing, and it may provide new clinical insights into acquired PRCA. Dysregulation of T cells has been shown to play a central role in PRCA. We studied T cell clonalities and STAT3 mutations in 90 PRCA patients and discovered that clonal T cell expansions were frequently recognized and closely associated with STAT3 mutations in the three major types of PRCA.

A novel germline GATA2 frameshift mutation with a premature stop codon in a family with congenital sensory hearing loss and myelodysplastic syndrome
International Journal of Hematology,114(2):286-291 2021(Aug.)
Author:Hideyuki Nakazawa; Tomomi Yamaguchi; Hitoshi Sakai; Masae Maruyama; Toru Kawakami; Fumihiro Kawakami; Sayaka Nishina; Masumi Ishikawa; Tomoki Kosho; Fumihiro Ishida

Serum sphingomyelin species profile is altered in hematologic malignancies
CLINICA CHIMICA ACTA,514:29-33 2021(Mar.)
Author:Hori, Atsushi; Ishida, Fumihiro; Nakazawa, Hideyuki; Yamaura, Makoto; Morita, Sunao; Uehara, Takeshi; Honda, Takayuki; Hidaka, Hiroya
Abstract:Sphingomyelin (SM) plays key roles in regulating cell membrane fluidity and in intracellular signal transduction. However, little is known as to whether alterations in SM concentration or SM species distribution are linked pathological conditions. The present study examined SM concentrations and species profiles in serum taken from patients with hematologic malignancies. Serum was collected from normal subjects and from patients with B-cell lymphoma, myelodysplastic syndrome (MDS), acute myeloid leukemia (AML) and acute lymphatic leukemia/ lymphoblastic lymphoma (ALL/LBL). Serum SM species distribution was analyzed using electrospray ionization mass spectrometry/ mass spectrometry (ESI MS/MS). Serum lipids concentration were measured using enzymatic assays. Normal and hematologic malignancy sera were similar in terms of total serum SM and phosphatidylcholine (PC) concentrations and SM/PC ratio. However, all hematologic malignancy sera had lower levels of SM species containing saturated odd chain fatty acids (OCFAs) in the side chain compared to normal serum. In addition, the proportion of SM species with saturated (C20 and C22) and mono unsaturated fatty acids (C18, C20, C22) were lower in MDS patient serum compared to normal serum. The present study revealed that the serum SM species profile in patients with hematologic malignancies differed from that of normal subjects despite total serum SM and PC concentrations and SM/PC ratios being similar between the various cancer groups and the normal group.

Allogeneic stem cell transplantation for patients with aggressive NK-cell leukemia.
Bone marrow transplantation,56(2):347-356 2021(Feb.)
Author:Ayumi Fujimoto; Fumihiro Ishida; Koji Izutsu; Satoshi Yamasaki; Dai Chihara; Junji Suzumiya; Tetsuo Mitsui; Noriko Doki; Hitoshi Sakai; Hikaru Kobayashi; Junya Kanda; Takahiro Fukuda; Yoshiko Atsuta; Ritsuro Suzuki
Abstract:Aggressive NK-cell leukemia (ANKL) has a fulminant clinical course with a poor prognosis. Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is currently the only curative treatment. Using the Japanese transplant registry data, the outcomes of 59 ANKL patients who underwent first allo-HSCT were analyzed. Twenty-nine patients received stem cells from cord blood (CB), 18 from peripheral blood, and 12 from bone marrow. At the time of transplant 21 patients had complete response (CR), and 7 partial response (PR), but 31 without response. The 1-year and 5-year overall survival (OS) were 33.9% and 27.3%, respectively. The 1-year cumulative incidences of relapse or progression was 55.5%, and that of non-relapse mortality was 12.1%. The OS was significantly better for patients with CR or PR at the time of allo-HSCT (P = 0.046), which was equivalent to that for patients who experienced primary induction failure at the time of allo-HSCT but achieved CR afterwards (40.6% versus 32.0% at 5 years; P = 0.95). Patients receiving CB had a significantly better OS than those receiving stem cells from others (37.3% versus 16.2% at 5 years; P = 0.04). Patients achieving event-free survival at 12 months after allo-HSCT had good outcomes with 5-year OS of 85.2%.

CCR4-positive peripheral T-cell lymphoma presenting as eosinophilic pneumonia and developing from prolonged pustular psoriasis
JOURNAL OF INTERNATIONAL MEDICAL RESEARCH,49(2):0300060521996165-300060521996165 2021(Feb.)
Author:Sekiguchi, Nodoka; Komatsu, Masamichi; Ichiyama, Takashi; Kobayashi, Aya; Gomi, Daisuke; Fukushima, Toshirou; Kobayashi, Takashi; Noguchi, Takuro; Nakazawa, Hideyuki; Asano, Naoko; Ishida, Fumihiro; Koizumi, Tomonobu
Abstract:A 29-year-old woman with chronic, prolonged pustular psoriasis was admitted to our hospital because of high-grade fever and a systemic skin rash. General examination revealed a whole-body skin rash and superficial lymphadenopathy. Peripheral blood examination showed unclassified cells positive for CD3, CD4, and T-cell receptor αβ, and negative for CD20 and CD56. Soon after administration, she developed acute respiratory failure and required artificial ventilation. Bronchoalveolar lavage fluid showed increased numbers of eosinophils and abnormal lymphocytes of the same phenotype in peripheral blood and skin. She was diagnosed with eosinophilic pneumonia, and her respiratory failure was improved by corticosteroid therapy. Based on the histological findings of skin, lymph node, and bone marrow biopsies, a diagnosis of peripheral T-cell lymphoma not otherwise specified (PTCL-NOS), with positivity for CC chemokine receptor 4 was made. She received chemotherapy followed by allogeneic stem cell transplantation, which resulted in complete remission of her PTCL-NOS. She remained alive and disease-free 6 years later. This is the first reported case of PTCL-NOS developing during the clinical course of pustular psoriasis. The clinical manifestations of PTCL-NOS are complex, but an accurate diagnosis and appropriate therapy may produce a good clinical outcome in patients with PTCL-NOS.

Isolated splenic Mycobacterium tuberculosis complex infection in an immunocompetent individual with FDG-PET positive mass
Journal of Infection and Chemotherapy,27(2):354-358 2021(Feb.)
Author:Sayaka Nishina; Hitoshi Sakai; Toru Kawakami; Shinichiro Kanai; Atsuhito Ushiki; Tatsuya Natori; Yuriko Igarashi; Satoshi Mitarai; Takashi Yoshiyama; Fumihiro Ishida; Hideyuki Nakazawa

Clonal hematopoiesis in adult pure red cell aplasia
SCIENTIFIC REPORTS,11(1):2253-2253 2021
Author:Fujishima, Naohito; Kohmaru, Junki; Koyota, Souichi; Kuba, Keiji; Saga, Tomoo; Omokawa, Ayumi; Moritoki, Yuki; Ueki, Shigeharu; Ishida, Fumihiro; Nakao, Shinji; Matsuda, Akira; Ohta, Akiko; Tohyama, Kaoru; Yamasaki, Hiroshi; Usuki, Kensuke; Nakashima, Yasuhiro; Sato, Shinya; Miyazaki, Yasushi; Nannya, Yasuhito; Ogawa, Seishi; Sawada, Kenichi; Mitani, Kinuko; Hirokawa, Makoto
Abstract:Idiopathic pure red cell aplasia (PRCA) and secondary PRCA associated with thymoma and large granular lymphocyte leukemia are generally considered to be immune-mediated. The PRCA2004/2006 study showed that poor responses to immunosuppression and anemia relapse were associated with death. PRCA may represent the prodrome to MDS. Thus, clonal hematopoiesis may be responsible for treatment failure. We investigated gene mutations in myeloid neoplasm-associated genes in acquired PRCA. We identified 21 mutations affecting amino acid sequences in 11 of the 38 adult PRCA patients (28.9%) using stringent filtering of the error-prone sequences and SNPs. Four PRCA patients showed 7 driver mutations in TET2, DNMT3A and KDM6A, and 2 PRCA patients carried multiple mutations in TET2. Five PRCA patients had mutations with high VAFs exceeding 0.3. These results suggest that clonal hematopoiesis by stem/progenitor cells might be related to the pathophysiology of chronic PRCA in certain adult patients.

T(o) be, or (not) to B, or both? Somatically mutated clonal T cells in common variable immunodeficiency and related immunodeficiencies
HAEMATOLOGICA,105(12):2702-2703 2020(Dec.)
Author:Ishida, Fumihiro; Nakazawa, Hideyuki

Primary thyroid T‐cell lymphoma with leukemic manifestation
eJHaem,1(1):18-18 2020(Jul.)
Author:Toru Kawakami; Hideyuki Nakazawa; Fumihiro Ishida

Magnetic Resonance Imaging-negative, Rituximab-resistant Neurolymphomatosis as a Paradoxical Presentation of Relapsed Primary Adrenal Lymphoma.
Internal medicine (Tokyo, Japan),59(11):1437-1443 2020(Jun. 01)
Author:Mayuka Nishikawara; Toru Kawakami; Hitoshi Sakai; Fumihiro Kawakami; Sayaka Nishina; Takeshi Uehara; Fumihiro Ishida; Hideyuki Nakazawa
Abstract:Primary adrenal lymphoma (PAL) is rare and known to have a predilection for central nervous system (CNS) relapse. A 70-year-old man with a 2-year history of primary aldosteronism presented because of a fever. He was hypotensive, and his adrenal glands were unequivocally enlarged. PAL was diagnosed. Despite showing an initial response to immunochemotherapy, progressive paralysis ensued. Magnetic resonance imaging findings were negative, and rituximab was ineffective. His debilitated condition hindered further chemotherapy. A postmortem examination revealed lymphoma relapse in the systemic peripheral nerves. The sequential presentation of two rare lymphomas implies that PAL might have a predilection for not only the CNS but also peripheral nerves.

Magnesium-dependent activated partial thromboplastin time assay-Simple method for lupus anticoagulant detection
Author:Tokutake, Takayoshi; Ieko, Masahiro; Naito, Sumiyoshi; Yoshida, Mika; Baba, Hisami; Kobayashi, Hikaru; Ishida, Fumihiro

High frequency of STAT3 gene mutations in T-cell receptor (TCR)gamma delta-type T-cell large granular lymphocytic leukaemia: implications for molecular diagnostics
Author:Yamane, Taku; Kawakami, Toru; Sekiguchi, Nodoka; Kobayashi, Jun; Ueki, Toshimitsu; Kobayashi, Hikaru; Kawakami, Fumihiro; Nishina, Sayaka; Sakai, Hitoshi; Oshimi, Kazuo; Higuchi, Yumiko; Nakazawa, Hideyuki; Ishida, Fumihiro

[Recent progress in the diagnosis and management of acquired pure red cell aplasia].
[Rinsho ketsueki] The Japanese journal of clinical hematology,61(9):1098-1104 2020
Author:Fumihiro Ishida
Abstract:Acquired pure red cell aplasia (PRCA) is characterized by normocytic anemia, reticulocytopenia, and a marked decrease in erythroid cell count in the bone marrow. PRCA develops in the context of various backgrounds, including recently recognized immune checkpoint inhibitor-associated PRCA, that need careful differential diagnoses. Besides humoral abnormalities such as major ABO-incompatible allogeneic hematopoietic stem cell transplantation-related PRCA, dysregulations of T cells have been shown. STAT3 gene mutations of cytotoxic T cells were identified in 40% of PRCA patients, which might suggest their use as novel molecular markers for PRCA. As initial management options for PRCA, red blood cell transfusion and immunosuppressive therapy (IST) drugs, such as cyclosporin, are usually selected. Roughly 80% of patients respond to IST; however, some relapse afterward or are refractory to IST. When patients with PRCA become refractory to two or three lines of IST, allogeneic hematopoietic stem cell transplantation (HCT) would become an appropriate choice, although the optimal procedures for allogeneic HCT have not been determined. A prospective study of PRCA in Japan has been ongoing since 2016 to solve the myriad clinical issues of PRCA.

臨床血液,61(9):1098-1104 2020
Author:石田 文宏
Keywords:Erythropoiesis;Cytotoxic T cell;Immunosuppressive therapy;STAT3

STAT3 mutations in natural killer cells are associated with cytopenia in patients with chronic lymphoproliferative disorder of natural killer cells
Author:Kawakami, Toru; Sekiguchi, Nodoka; Kobayashi, Jun; Yamane, Taku; Nishina, Sayaka; Sakai, Hitoshi; Hirabayashi, Yukio; Nakazawa, Hideyuki; Ishida, Fumihiro

Phase II study of FLAGM (fludarabine plus high-dose cytarabine plus granulocyte colony-stimulating factor plus mitoxantrone) for relapsed or refractory acute myeloid leukemia
Author:Hatsumi, Nahoko; Miyawaki, Shuichi; Yamauchi, Takahiro; Takeshita, Akihiro; Komatsu, Norio; Usui, Noriko; Arai, Yukihiro; Ishida, Fumihiro; Morii, Takeshi; Kano, Yasuhiko; Ogura, Michinori; Machida, Shinichiro; Nishii, Kazuhiro; Honda, Sumihisa; Ohnishi, Kazunori; Naoe, Tomoki; JALSG;

Tumor long-axis diameter and SUVmax predict long-term responders in 90Y-ibritumomab tiuxetan monotherapy
Author:Tsukamoto, Norifumi; Yokohama, Akihiko; Higuchi, Tetsuya; Mitsui, Takeki; Koiso, Hiromi; Takizawa, Makiko; Shimizu, Hiroaki; Ishizaki, Takuma; Matsumoto, Morio; Toyama, Kohtaro; Sakura, Tohru; Ogura, Hidemi; Saitoh, Takayuki; Ishida, Fumihiro; Murakami, Hirokazu; Tsushima, Yoshito; Handa, Hiroshi;

T-cell large granular lymphocytic (LGL) leukemia consists of CD4+/CD8dim and CD4-/CD8+ LGL populations in association with immune thrombocytopenia, autoimmune neutropenia, and monoclonal B-cell lymphocytosis.
Journal of clinical and experimental hematopathology : JCEH,59(4):202-206 2019
Author:Naoya Kuwahara; Taiichi Kodaka; Yuriko Zushi; Miho Sasaki; Takae Goka; Hayato Maruoka; Yumi Aoyama; Hiroko Tsunemine; Taku Yamane; Jun Kobayashi; Toru Kawakami; Fumihiro Ishida; Tomoo Itoh; Takayuki Takahashi
Abstract:CD3+/CD57+ T-cell large granular lymphocyte leukemia (T-LGLL) is an indolent neoplasm, exhibiting mostly CD8+, less frequently CD4+ phenotypes, and T-LGLL consisting of 2 populations with CD8+ and CD4+ phenotypes is markedly rare. An 87-year-old female was admitted under a diagnosis of immune thrombocytopenia (ITP) with a platelet count of 5.0×109/L and increased number of LGL with unknown etiology. Her neutrophil count also decreased to 0.27×109/L and she was positive for antineutrophil antibody. The WBC count was 2.7×109/L with 34.7% LGL and flow cytometry (FCM) analysis revealed 16% CD3+/CD4+/CD8dim/CD57+ and 20.9% CD3+/CD8+/CD57+ populations. These populations also expressed granzyme B and perforin. Circulating mononuclear cells were found to be clonal by PCR analysis of T-cell receptor β-chain gene. Serum immunofixation and bone marrow FCM analyses demonstrated 2 clonal B-cells producing IgG-λ and IgA-λ. Deep amplicon sequencing of STAT3 and STAT5B genes revealed a STAT3 R302G mutation with an allele burden of 2.6%. The thrombocytopenia and neutropenia were successfully treated by prednisolone and romiplostim with negative conversion of antineutrophil antibody. This is the first reported case of T-LGLL with dual components of CD4+/CD8dim and CD4-/CD8+ populations in terms of multiple comorbidities related to the respective CD8+ and CD4+ T-LGLLs.

Aggressive natural killer-cell leukemia mutational landscape and drug profiling highlight JAK-STAT signaling as therapeutic target
Nature Communications,9(1):1-12 2018(Dec. 01)
Author:Olli Dufva; Matti Kankainen; Tiina Kelkka; Nodoka Sekiguchi; Shady Adnan Awad; Samuli Eldfors; Bhagwan Yadav; Heikki Kuusanmäki; Disha Malani; Emma I Andersson; Paavo Pietarinen; Leena Saikko; Panu E. Kovanen; Teija Ojala; Dean A. Lee; Thomas P. Loughran; Hideyuki Nakazawa; Junji Suzumiya; Ritsuro Suzuki; Young Hyeh Ko; Won Seog Kim; Shih-Sung Chuang; Tero Aittokallio; Wing C. Chan; Koichi Ohshima; Fumihiro Ishida; Satu Mustjoki
Abstract:Aggressive natural killer-cell (NK-cell) leukemia (ANKL) is an extremely aggressive malignancy with dismal prognosis and lack of targeted therapies. Here, we elucidate the molecular pathogenesis of ANKL using a combination of genomic and drug sensitivity profiling. We study 14 ANKL patients using whole-exome sequencing (WES) and identify mutations in STAT3 (21%) and RAS-MAPK pathway genes (21%) as well as in DDX3X (29%) and epigenetic modifiers (50%). Additional alterations include JAK-STAT copy gains and tyrosine phosphatase mutations, which we show recurrent also in extranodal NK/T-cell lymphoma, nasal type (NKTCL) through integration of public genomic data. Drug sensitivity profiling further demonstrates the role of the JAK-STAT pathway in the pathogenesis of NK-cell malignancies, identifying NK cells to be highly sensitive to JAK and BCL2 inhibition compared to other hematopoietic cell lineages. Our results provide insight into ANKL genetics and a framework for application of targeted therapies in NK-cell malignancies.

Frequent STAT3 mutations in CD8+ T cells from patients with pure red cell aplasia
BLOOD ADVANCES,2(20):2704-2712 2018(Oct.)
Author:Toru Kawakami; Nodoka Sekiguchi; Jun Kobayashi; Tatsuya Imi; Kazuyuki Matsuda; Taku Yamane; Sayaka Nishina; Yasushi Senoo; Hitoshi Sakai; Toshiro Ito; Tomonobu Koizumi; Makoto Hirokawa; Shinji Nakao; Hideyuki Nakazawa; Fumihiro Ishida

Aggressive NK-Cell Leukemia
Author:Ishida, Fumihiro

Breakthrough Candida guilliermondii (Meyerozyma guilliermondii) fungemia after cord blood transplantation for extranodal NK-cell lymphoma with azole prophylaxis.
Transplant infectious disease : an official journal of the Transplantation Society,:e12922 2018(May)
Author:Nakazawa Hideyuki; Nishina Sayaka; Senoo Yasushi; Sakai Hitoshi; Ito Toshiro; Kikuchi Ken; Ishida Fumihiro
Abstract:Fluconazole (FLCZ) is an azole antifungal agent and it has shown excellent clinical activities in suppressing fungemia with Candida albicans after hematopoietic stem cell transplantation. Increased administration of prophylactic FLCZ seems to have given rise to the relatively higher incidence of more resistant Candida non-albicans infection. We present a case with a rare breakthrough fungemia with C. guilliermondii after cord blood transplantation for Extranodal NK cell Lymphoma, nasal type (ENKL), during antifungal prophylaxis with FLCZ. High level of caution is needed for the breakthrough, especially after long- term azole administration.

Severe infection of pseudomonas aeruginosa during eculizumab therapy for paroxysmal nocturnal hemoglobinuria
Internal Medicine,57(1):127-130 2018
Author:Toru Kawakami; Hideyuki Nakazawa; Yukifumi Kurasawa; Hitoshi Sakai; Sayaka Nishina; Noriko Senoo; Yasushi Senoo; Fumihiro Ishida
Abstract:Eculizumab is the complement inhibitor administered to ameliorate intravascular hemolysis in paroxysmal nocturnal hemoglobinuria. Whether or not the inhibitory mechanism may also increase the susceptibility to non-Neisserial infection is unclear. A 73-year old woman presented with bacteremia, cholecystitis and liver abscess with Pseudomonas aeruginosa. Although she had been neutropenic for 21 years, she had no history of severe infection before eculizumab had been administered. The infection with P. aeruginosa was successfully controlled with antibiotics, granulocyte colony-stimulating factor and cholecystectomy. The present case might be representative of less common bacterial infections than Neisseria spp. among patients treated with eculizumab.

Successful treatment with vitamin B6 for X-linked sideroblastic anemia with ALAS2 R452H mutation.
Rinsho Ketsueki.,59:401-406 2018
Author:Kawakami T; Nakazawa H; Kawakami F; Matsuzawa S; Sudo Y; Sakai H; Nishina S; Sendo N; Sendo Y; Komatsu M; Umemura T; Yamaguti T; Kosho T; Fujiwara T; Harigae H; Ishida F

大顆粒リンパ球増多症 (特集 リンパ球の増減を正しく評価するために)
臨床検査 = Journal of clinical laboratory medicine,61(8):976-982 2017(Aug.)
Author:石田 文宏
Abstract:臨床検査 = Journal of clinical laboratory medicine

Oral cyclophosphamide was effective for Coombs-negative autoimmune hemolytic anemia in CD16+CD56-chronic lymphoproliferative disorder of NK-cells
Author:Nodoka Sekiguchi; Sayaka Nishina; Toru Kawakami; Hitoshi Sakai; Noriko Senoo; Yasushi Senoo; Toshiro Ito; Hiroshi Saito; Hideyuki Nakazawa; Tomonobu Koizumi; Fumihiro Ishida
Abstract:An 84-year-old woman was referred to our hospital presenting anemia. Her hemoglobin level was 5.8 g/dL, and white blood cell count was 9400/mu L, consisting of 82% lymphocytes. Given the lymphocyte phenotype (CD2+, CD3-, CD16+, and CD56-) and negative whole blood EBV viral load, we made a diagnosis of chronic lymphoproliferative disorder of NK cells (CLPD-NK). We suspected hemolytic anemia because of the high levels of reticulocytes in the peripheral blood and the low haptoglobin value. Although the direct Coombs test was negative and there was no cold agglutination, we examined her red-blood-cell-bound IgG (RBC-IgG), which was elevated. She was diagnosed as having as Coombs-negative autoimmune hemolytic anemia (AIHA). We report the effectiveness of oral cyclophosphamide for Coombs-negative autoimmune hemolytic anemia in CLPD-NK.

A screening method with lymphocyte percentage and proportion of granular lymphocytes in the peripheral blood for large granular lymphocyte (LGL) leukemia
Author:Takahiro Tanahashi; Nodoka Sekiguchi; Kazuyuki Matsuda; Akihiro Matsumoto; Toshiro Ito; Hideyuki Nakazawa; Fumihiro Ishida
Abstract:Large granular lymphocyte leukemia (LGL-L) is defined morphologically as a group of lymphoproliferative disorders, including T-cell large granular lymphocytic leukemia (T-LGL-L), chronic lymphoproliferative disorders of NK cells (CLPD-NK), and aggressive NK cell leukemia. We investigated the morphological features of LGL leukemic cells in LGL-L patients to identify screening methods best suited to application in daily clinical practice. LGL leukemic cells were mostly indistinguishable from normal LGL; however, we developed a simplified approach to distinguishing among these cells, in which lymphocyte % and the proportion of granular lymphocytes among lymphocytes (GL %) can serve as parameters at the cut-off values of 52 and 50 %, respectively. We confirmed the accuracy of these methods in validation groups. It may be useful to evaluate GL % to screen for LGL-L using a cut-off threshold of 50 when Lym % exceeds 52 in the peripheral blood examination in cases in which LGL-L is suspected in daily practice while keeping clinical context and the coefficient of variation of these parameters under consideration.

節外性NK細胞リンパ腫との鑑別が問題となったplasmablastic lymphoma
臨床血液,57(11):2426-2427 2016(Nov.)
Author:松澤 周治; 川上 徹; 須藤 裕里子; 酒井 均; 仁科 さやか; 妹尾 紀子; 小原 洋一; 佐野 健司; 浅野 直子; 石田 文宏; 中澤 英之

High incidence of activating STAT5B mutations in CD4-positive T-cell large granular lymphocyte leukemia
BLOOD,128(20):2465-2468 2016(Nov.)
Author:Emma I. Andersson; Takahiro Tanahashi; Nodoka Sekiguchi; Vanessa Rebecca Gasparini; Sabrina Bortoluzzi; Toru Kawakami; Kazuyuki Matsuda; Takeki Mitsui; Samuli Eldfors; Stefania Bortoluzzi; Alessandro Coppe; Andrea Binatti; Sonja Lagstrom; Pekka Ellonen; Noriyasu Fukushima; Sayaka Nishina; Noriko Senoo; Hitoshi Sakai; Hideyuki Nakazawa; Yok-Lam Kwong; Thomas P. Loughran; Jaroslaw P. Maciejewski; Satu Mustjoki; Fumihiro Ishida

Cell size variations of large granular lymphocyte leukemia: Implication of a small cell subtype of granular lymphocyte leukemia with STAT3 mutations
LEUKEMIA RESEARCH,45:8-13 2016(Jun.)
Author:Takahiro Tanahashi; Nodoka Sekiguchi; Kazuyuki Matsuda; Yuka Takezawa; Toshiro Ito; Hikaru Kobayashi; Naoaki Ichikawa; Sayaka Nishina; Noriko Senoo; Hitoshi Sakai; Hideyuki Nakazawa; Fumihiro Ishida
Abstract:Large granular lymphocyte leukemia (LGL-L) has been morphologically defined as a group of lymphoproliferative disorders, including T-cell large granular lymphocytic leukemia (T-LGL-L), chronic lymphoproliferative disorders of NK cells (CLPD-NK) and aggressive NK cell leukemia. We investigated the morphological features of LGL leukemic cells in 26 LGL-L patients in order to elucidate relationships with current classifications and molecular backgrounds. LGL-L cells were mostly indistinguishable from normal LGL. Patients with STAT3 SH2 domain mutations showed significantly smaller cells compared with patients without STAT3 mutations. Four patients with T-LGL-L showed smaller granular lymphocytes with a median diameter of less than 13 mu m, which were rarely seen in normal subjects. This small subtype of T-LGL-L was recognized among rather young patients and was associated with D661Y mutations in the STAT3 gene SH2 domain. In addition, all of them showed anemia including two cases with pure red cell aplasia. These results suggest the heterogeneity of T-LGL-L and a specific subtype with small variants of T-LGL-L. (C) 2016 Elsevier Ltd. All rights reserved.

Exogenous Magnesium Chloride Reduces the Activated Partial Thromboplastin Times of Lupus Anticoagulant-Positive Patients
PLOS ONE,11(6):- 2016(Jun.)
Author:Takayoshi Tokutake; Hisami Baba; Yuji Shimada; Wataru Takeda; Keijiro Sato; Yuki Hiroshima; Takehiko Kirihara; Ikuo Shimizu; Hideyuki Nakazawa; Hikaru Kobayashi; Fumihiro Ishida
Abstract:The activated partial thromboplastin time (APTT) assay is a basic hemostatic assay based on the time it takes for clots to form in plasma samples after the addition of calcium chloride. It is used to screen for various coagulation disorders. Several previous reports have suggested that magnesium (Mg) might contribute to coagulation reactions by binding to specific coagulation proteins. We investigated the effects of Mg on the APTT. In healthy controls, the APTT was significantly prolonged in proportion to the increase in the concentration of magnesium chloride in the range from 2.1 to 16.7 mmol/L. Among eight samples from patients with various disorders that exhibited prolonged APTT, two samples demonstrated shorter APTT when Mg was added, both of which were from patients that were positive for lupus anticoagulant. When we examined 206 clinical APTT samples, we found that Mg shortened the APTT of two samples. These two samples were also from lupus anticoagulant-positive patients (p-value: < 0.003). Our findings regarding the unique effects of exogenous Mg on the APTT of lupus anticoagulant-positive patients might shed light on the role of Mg in APTT assays and lead to the development of a novel screening method for lupus anticoagulant.

A prognostic index for natural killer cell lymphoma after non-anthracycline-based treatment: a multicentre, retrospective analysis
LANCET ONCOLOGY,17(3):389-400 2016(Mar.)
Author:Seok Jin Kim; Dok Hyun Yoon; Arnaud Jaccard; Wee Joo Chng; Soon Thye Lim; Huangming Hong; Yong Park; Kian Meng Chang; Yoshinobu Maeda; Fumihiro Ishida; Dong-Yeop Shin; Jin Seok Kim; Seong Hyun Jeong; Deok-Hwan Yang; Jae-Cheol Jo; Gyeong-Won Lee; Chul Won Choi; Won-Sik Lee; Tsai-Yun Chen; Kiyeun Kim; Sin-Ho Jung; Tohru Murayama; Yasuhiro Oki; Ranjana Advani; Francesco d'Amore; Norbert Schmitz; Cheolwon Suh; Ritsuro Suzuki; Yok Lam Kwong; Tong-Yu Lin; Won Seog Kim
Abstract:Background The clinical outcome of extranodal natural killer T-cell lymphoma (ENKTL) has improved substantially as a result of new treatment strategies with non-anthracycline-based chemotherapies and upfront use of concurrent chemoradiotherapy or radiotherapy. A new prognostic model based on the outcomes obtained with these contemporary treatments was warranted. Methods We did a retrospective study of patients with newly diagnosed ENKTL without any previous treatment history for the disease who were given non-anthracycline-based chemotherapies with or without upfront concurrent chemoradiotherapy or radiotherapy with curative intent. A prognostic model to predict overall survival and progression-free survival on the basis of pretreatment clinical and laboratory characteristics was developed by filling a multivariable model on the basis of the dataset with complete data for the selected risk factors for an unbiased prediction model. The final model was applied to the patients who had complete data for the selected risk factors. We did a validation analysis of the prognostic model in an independent cohort. Findings We did multivariate analyses of 527 patients who were included from 38 hospitals in 11 countries in the training cohort. Analyses showed that age greater than 60 years, stage III or IV disease, distant lymph-node involvement, and non-nasal type disease were significantly associated with overall survival and progression-free survival. We used these data as the basis for the prognostic index of natural killer lymphoma (PINK), in which patients are stratified into low-risk (no risk factors), intermediate-risk (one risk factor), or high-risk (two or more risk factors) groups, which were associated with 3-year overall survival of 81% (95% CI 75-86), 62% (55-70), and 25% (20-34), respectively. In the 328 patients with data for Epstein-Barr virus DNA, a detectable viral DNA titre was an independent prognostic factor for overall survival. When these data were added to PINK as the basis for another prognostic index (PINK-E)-which had similar low-risk (zero or one risk factor), intermediate-risk (two risk factors), and high-risk (three or more risk factors) categories-significant associations with overall survival were noted (81% [95% CI 75-87%], 55% (44-66), and 28% (18-40%), respectively). These results were validated and confirmed in an independent cohort, although the PINK-E model was only significantly associated with the high-risk group compared with the low-risk group. Interpretation PINK and PINK-E are new prognostic models that can be used to develop risk-adapted treatment approaches for patients with ENKTL being treated in the contemporary era of non-anthracycline-based therapy.

NK細胞腫瘍の診断と治療における最近の進歩 (第76回日本血液学会学術集会) -- (Symposium : Advances in the treatment of NK-cell malignancies)
臨床血液,56(6):645-650 2015(Jun.)
Author:石田 文宏

Evaluation of Epstein-Barr virus DNA loads in peripheral blood using a plasmid solution calibrated with the 1st WHO International Standard
Japanese Journal of Medical Technology,64(1):48-53 2015
Author:IWASHITA Chinami; SUEKI Akane; MATSUDA Kazuyuki; IDE Yuichiro; SUGANO Mitsutoshi; ISHIDA Fumihiro; HONDA Takayuki
Abstract:The monitoring of Epstein-Barr virus (EBV) DNA loads is important for the detection and prevention of EBV-related disorders. EBV loads have been evaluated by quantitative real-time PCR analysis requiring the preparation of standards. The standard materials for the quantification of EBV loads vary among clinical laboratories. Therefore, it is difficult to obtain comparable EBV DNA quantitative values among laboratories. In this study, the copy number of plasmids containing a fragment corresponding to the EBV BNRF1 gene was calibrated with the 1st WHO International Standard for EBV quantification tests, and then the calibrated plasmids were used as secondary standards to quantitate the EBV DNA loads of subsequent samples from patients. Additionally, the theoretical conversion factor was calculated to convert the EBV DNA loads obtained using noncalibrated standards into those obtained using the present secondary standards. The EBV DNA loads in hematopoietic-stem-cell-transplanted patients suspected of having post-transplant lymphoproliferative disorder (PTLD) and patients with EBV-related hemophagocytic lymphohistiocytosis (EBV-HLH) were 8.74 × 102−1.51 × 104 and 1.58 × 104−1.33 × 106 IU/μg DNA, respectively, as determined by these methods. The utilization of secondary unified standards for the quantification of EBV DNA loads available for many clinical laboratories would contribute to the establishment of a diagnostic reference and therapeutic intervention criteria based on the quantitative values of EBV DNA, not only in PTLD patients but also in those with various disorders related to EBV infection.

Prognostic significance of pleural or pericardial effusion and the implication of optimal treatment in primary mediastinal large B-cell lymphoma: a multicenter retrospective study in Japan
HAEMATOLOGICA,99(12):1817-1825 2014(Dec.)
Author:Tomohiro Aoki; Koji Izutsu; Ritsuro Suzuki; Chiaki Nakaseko; Hiroshi Arima; Kazuyuki Shimada; Akihiro Tomita; Makoto Sasaki; Jun Takizawa; Kinuko Mitani; Tadahiko Igarashi; Yoshinobu Maeda; Noriko Fukuhara; Fumihiro Ishida; Nozomi Niitsu; Ken Ohmachi; Hirotaka Takasaki; Naoya Nakamura; Tomohiro Kinoshita; Shigeo Nakamura; Michinori Ogura
Abstract:The prognosis of patients with primary mediastinal large B-cell lymphoma has improved over recent years. However, the optimal treatment strategy including the role of radiotherapy remains unknown. We retrospectively analyzed the clinical outcomes of 345 patients with newly diagnosed primary mediastinal large B-cell lymphoma in Japan. With a median follow up of 48 months, the overall survival at four years for patients treated with R-CHOP (n=187), CHOP (n=44), DA-EPOCH-R (n=9), 2nd- or 3rd-generation regimens, and chemotherapy followed by autologous stem cell transplantation were 90%, 67%, 100%, 91% and 92%, respectively. Focusing on patients treated with R-CHOP, a higher International Prognostic Index score and the presence of pleural or pericardial effusion were identified as adverse prognostic factors for overall survival in patients treated with R-CHOP without consolidative radiotherapy (IPI: hazard ratio 4.23, 95% confidence interval 1.48-12.13, P=0.007; effusion: hazard ratio 4.93, 95% confidence interval 1.37-17.69, P=0.015). Combined with the International Prognostic Index score and the presence of pleural or pericardial effusion for the stratification of patients treated with R-CHOP without radiotherapy, patients with lower International Prognostic Index score and the absence of effusion comprised approximately one-half of these patients and could be identified as curable patients (95% overall survival at 4 years). The DA-EPOCH-R regimen might overcome the effect of these adverse prognostic factors. Our simple indicators of International Prognostic Index score and the presence of pleural or pericardial effusion could stratify patients with primary mediastinal large B-cell lymphoma and help guide selection of treatment.

STAT3 gene mutations and their association with pure red cell aplasia in large granular lymphocyte leukemia
CANCER SCIENCE,105(3):342-346 2014(Mar.)
Author:Fumihiro Ishida; Kazuyuki Matsuda; Nodoka Sekiguchi; Hideki Makishima; Chiaki Taira; Kayoko Momose; Sayaka Nishina; Noriko Senoo; Hitoshi Sakai; Toshiro Ito; Yok-Lam Kwong
Abstract:Large granular lymphocyte leukemia (LGLL) has been morphologically characterized as a group of lymphoproliferative diseases that include T-cell large granular lymphocytic leukemia (T-LGLL) and chronic lymphoproliferative disorders of natural killer cells (CLPD-NK). We investigated mutations in the Src homology 2 (SH2) domain of the signal transducer and activator of transcription 3 (STAT3) gene in Asian cohorts of T-LGLL and CLPD-NK (n=42 and 11, respectively). Two mutations, Y640F and D661Y, were identified using direct sequencing or allele-specific (AS) PCR. Y640F and D661Y mutations were found in seven and 18 patients, respectively. Two patients were positive for both mutations. Frequencies of STAT3 mutations in T-LGLL and CLPD-NK were 47.6% and 27.2%, respectively. Pure red cell aplasia (PRCA) was associated with the mutations (P=0.005). The mutations were persistently found at stable levels in some patients after more than 5years using AS-quantitative PCR. The results of the present study indicate that the SH2 domain of the STAT3 gene is frequently mutated in Asian T-LGLL and CLPD-NK, and that PRCA is closely correlated with the mutations. SH2 domain of the STAT3 gene is frequently mutated in Asian T cell large granular lymphocyte leukemia and chronic lymphoproliferative disorders of NK cells. Pure red cell aplasia is closely associated with the mutations.

Screening Tests Using Serum Tissue Transglutaminase IgA May Facilitate the Identification of Undiagnosed Celiac Disease among Japanese Population
Author:Hideyuki Nakazawa; Hideki Makishima; Toshiro Ito; Hiroyoshi Ota; Kayoko Momose; Nodoka Sekiguchi; Kaname Yoshizawa; Taiji Akamatsu; Fumihiro Ishida
Abstract:The prevalence of celiac disease (CD) among Japanese population has been unknown, whereas it has been increasingly recognized in the US and in the European countries. The aim of the present study is to identify possible cases with CD among Japanese population and clarify the relevance of screening for the disease. We conducted a serologic screening for the disease among 710 Japanese patients and 239 healthy volunteers at a local tertiary teaching hospital, using an anti-tissue transglutaminase IgA (TTG-IgA) test, and histological examination of the small intestines from the TTG-IgA positive subjects. There were no TTG-IgA positive sera among the healthy volunteers. Twenty of the patients (2.8%), including eight with malignant lymphoma, were tested positive for TTG-IgA. The histological examination of the eleven patients among those with positive TTG-IgA, seven showed villous atrophy and partial lymphocytes infiltration in the mucosa, which could be compatible to mucosal changes observed in CD. Five of them had non-Hodgkin lymphoma in the gastrointestinal tracts. Serologic tests using TTG-IgA might be relevant to screen for those with undiagnosed CD among Japanese population.

Large vessel vasculitis developed early after allogeneic bone marrow transplant for acute erythroid leukemia
LEUKEMIA & LYMPHOMA,54(12):2748-2749 2013(Dec.)
Author:Toru Kawakami; Toshiro Ito; Hitoshi Sakai; Nodoka Sekiguchi; Hideyuki Nakazawa; Noriko Senoo; Sayaka Nishina; Fumihiro Ishida

Liver Dysfunction and Thrombocytopenia Diagnosed as Intravascular Large B-cell Lymphoma Using a Timely and Accurate Transjugular Liver Biopsy
INTERNAL MEDICINE,52(17):1903-1908 2013
Author:Nodoka Sekiguchi; Satoru Joshita; Toshikazu Yoshida; Masahiro Kurozumi; Kenji Sano; Michitaka Nakagawa; Tetsuya Ito; Tsuyoshi Matsushita; Daisuke Komatsu; Michiharu Komatsu; Toshiro Ito; Takeji Umemura; Shu-ichi Ikeda; Masumi Kadoya; Fumihiro Ishida; Eiji Tanaka
Abstract:A 72-year-old man suffered from paraparesis with a sensory impairment and bladder and rectal disturbances. Magnetic resonance imaging T2-weighted images depicted a high-intensity lesion in the spinal cord that was consistent with myelitis. A blood examination revealed severe thrombocytopenia and liver dysfunction. No malignant cells were detected by peripheral smears or bone marrow biopsy. Systemic computed tomography detected hepatosplenomegaly and ascites but no lymphadenopathies. Transjugular liver biopsy (TJLB) safely confirmed a diagnosis of intravascular large B-cell lymphoma (IVLBCL), and the patient achieved a complete response following treatment with an appropriate chemotherapy. TJLB is therefore a timely and accurate diagnostic approach for IVLBCL, especially when a bleeding tendency and ascites are noted.

Spliceosome-related gene mutations in myelodysplastic syndrome can be used as stable markers for monitoring minimal residual disease during follow-up
LEUKEMIA RESEARCH,36(11):1393-1397 2012(Nov.)
Author:Kazuyuki Matsuda; Fumihiro Ishida; Toshiro Ito; Hideyuki Nakazawa; Shuhei Miura; Chiaki Taira; Akane Sueki; Yukihiro Kobayashi; Takayuki Honda
Abstract:Various gene mutations have been reported in patients with myelodysplastic syndrome (MDS). Serial studies of mutations during follow-up are important for investigating the stability of the mutations for use as minimal residual disease (MRD) markers. Sequential quantitative analyses of 5 patients with spliceosome-related gene mutations by allele-specific quantitative polymerase chain reaction revealed that the U2AF1 S34F and SF3B1 K666N were persistently retained during the disease progression. The spliceosome-related gene mutations appear to be stable during disease progression and may be useful as potential markers for MRD monitoring in MDS patients that usually lack established specific MRD markers. (C) 2012 Elsevier Ltd. All rights reserved.

Elevated serum granulysin and its clinical relevance in mature NK-cell neoplasms
Author:Nodoka Sekiguchi; Naoko Asano; Toshiro Ito; Kayoko Momose; Masanobu Momose; Fumihiro Ishida
Abstract:Mature natural killer (NK)-cell neoplasms include extranodal NK/T cell lymphoma, nasal type (ENKL), aggressive NK-cell leukemia (ANKL) and chronic lymphoproliferative disorders of NK cells (CLPD-NK). Granulysin, a cytolytic granule protein, is expressed in cytotoxic T cells and NK cells, and is found in the sera as well, and functions as a cytotoxic and proinflammatory protein. Cytolytic proteins, such as granzyme B and perforin, have been shown to play crucial pathophysiological roles in NK/T cell neoplasms and have also been utilized for diagnostic purposes. Granulysin in NK-cell proliferative disorders, however, has yet to be fully analyzed. To elucidate the clinical relevance of granulysin in mature NK-cell neoplasms, we measured serum granulysin and analyzed cytolytic molecules immunohistologically. The median concentrations of serum granulysin were 39.0, 2.85, 2.8 and 1.35 ng/ml in ANKL, ENKL, CLPD-NK and healthy subjects, respectively (P < 0.01). Serum granulysin was significantly elevated in patients with ANKL compared with the levels in ENKL (P = 0.006) and CLPD-NK (P = 0.037). Furthermore, serum granulysin was correlated with whole-blood EBV viral load in ENKL and ANKL (P = 0.005) and was significantly reduced after treatment. Different expression patterns of cytolytic granule proteins were observed among the mature NK-cell neoplasms. Granulysin is closely associated with the characteristics of NK-cell neoplasms and serum granulysin may serve as a novel biomarker for these disorders.

Pretreatment EBV-DNA copy number is predictive of response and toxicities to SMILE chemotherapy for extranodal NK/T-cell lymphoma, nasal type.
Clinical cancer research : an official journal of the American Association for Cancer Research,18(15):4183-4190 2012(Aug.)
Author:Ito Y; Kimura H; Maeda Y; Hashimoto C; Ishida F; Izutsu K; Fukushima N; Isobe Y; Takizawa J; Hasegawa Y; Kobayashi H; Okamura S; Kobayashi H; Yamaguchi M; Suzumiya J; Hyo R; Nakamura S; Kawa K; Oshimi K; Suzuki R

Aggressive natural killer cell leukemia: Therapeutic potential of l-asparaginase and allogeneic hematopoietic stem cell transplantation
CANCER SCIENCE,103(6):1079-1083 2012(Jun.)
Author:Fumihiro Ishida; Young Hyeh Ko; Won Seog Kim; Junji Suzumiya; Yasushi Isobe; Kazuo Oshimi; Shigeo Nakamura; Ritsuro Suzuki
Abstract:We conducted a retrospective JapanKorea multicenter study to better elucidate the clinicopathologic features and therapeutic modalities for aggressive natural killer cell leukemia (ANKL). A total of 34 patients were analyzed. The median age of the patients was 40 years. Among the patients in the study, four had a history of EpsteinBarr virus-related disorders. Three types of ANKL cells were categorized according to their morphological features. Leukemic cells were below 20% in both peripheral blood and bone marrow of 11 patients. The clinical characteristics and prognoses of these 11 patients did not differ significantly from those of the others. As an initial therapy, l-asparaginase chemotherapy resulted in a better response. A total of six patients received allogeneic hematopoietic stem cell transplantation (HSCT) and two received autologous HSCT, with all in non-complete remission (CR). After HSCT, four with allogeneic and one with autologous HSCT reached CR. Median survival of all patients was 51 days. Median survival for the patients with and without HSCT were 266 and 36 days, respectively. A total of two patients with allogeneic HSCT were alive and in CR. All patients without HSCT died of ANKL. The use of l-asparaginase was indicated as a factor for longer survival (HR 0.33, 95% confidence interval; 0.130.83, P = 0.02). Early diagnosis of ANKL, l-asparaginase-based chemotherapy and allogeneic HSCT might lead to improved patient outcomes. (Cancer Sci 2012; 103: 10791083)

Long-term haematological improvement after non-intensive or no chemotherapy in juvenile myelomonocytic leukaemia and poor correlation with adult myelodysplasia spliceosome-related mutations
BRITISH JOURNAL OF HAEMATOLOGY,157(5):647-650 2012(Jun.)
Author:Matsuda, Kazuyuki; Yoshida, Nao; Miura, Shuhei; Nakazawa, Yozo; Sakashita, Kazuo; Hyakuna, Nobuyuki; Saito, Masahiro; Kato, Fumiyo; Ogawa, Atsushi; Watanabe, Akihiro; Sotomatsu, Manabu; Kobayashi, Chie; Ito, Toshiro; Ishida, Fumihiro; Manabe, Atsushi; Kojima, Seiji; Koike, Kenichi

Phase II Study of SMILE Chemotherapy for Newly Diagnosed Stage IV, Relapsed, or Refractory Extranodal Natural Killer (NK)/T-Cell Lymphoma, Nasal Type: The NK-Cell Tumor Study Group Study
JOURNAL OF CLINICAL ONCOLOGY,29(33):4410-4416 2011(Nov.)
Author:Motoko Yamaguchi; Yok-Lam Kwong; Won Seog Kim; Yoshinobu Maeda; Chizuko Hashimoto; Cheolwon Suh; Koji Izutsu; Fumihiro Ishida; Yasushi Isobe; Eisaburo Sueoka; Junji Suzumiya; Takao Kodama; Hiroshi Kimura; Rie Hyo; Shigeo Nakamura; Kazuo Oshimi; Ritsuro Suzuki
Abstract:Purpose To explore a more effective treatment for newly diagnosed stage IV, relapsed, or refractory extranodal natural killer/T-cell lymphoma, nasal type (ENKL), we conducted a phase II study of the steroid (dexamethasone), methotrexate, ifosfamide, L-asparaginase, and etoposide (SMILE) regimen. Patients and Methods Patients with newly diagnosed stage IV, relapsed, or refractory disease and a performance status of 0 to 2 were eligible. Two cycles of SMILE chemotherapy were administered as the protocol treatment. The primary end point was the overall response rate (ORR) after the protocol treatment. Results A total of 38 eligible patients were enrolled. The median age was 47 years (range, 16 to 67 years), and the male: female ratio was 21:17. The disease status was newly diagnosed stage IV in 20 patients, first relapse in 14 patients, and primary refractory in four patients. The eligibility was revised to include lymphocyte counts of 500/mu L or more because the first two patients died from infections. No treatment-related deaths were observed after the revision. The ORR and complete response rate after two cycles of SMILE chemotherapy were 79% (90% CI, 65% to 89%) and 45%, respectively. In the 28 patients who completed the protocol treatment, 19 underwent hematopoietic stem-cell transplantation. The 1-year overall survival rate was 55% (95% CI, 38% to 69%). Grade 4 neutropenia was observed in 92% of the patients. The most common grade 3 or 4 nonhematologic complication was infection (61%). Conclusion SMILE chemotherapy is an effective treatment for newly diagnosed stage IV, relapsed or refractory ENKL. Myelosuppression and infection during the treatment should be carefully managed.

Primary gastric diffuse large B-cell lymphoma with orbital involvement: diagnostic usefulness of 18F-fluorodeoxyglucose positron emission tomography.
Intern Med,50(18):1953-1956 2011(Sep.)
Author:Iwaya Y; Takenaka K; Akamatsu T; Yamada Y; Haba S; Matsuda K; Nakazawa H; Muraki T; Kaneko Y; Ito T; Sano K; Arakura N; Tanaka E; Ishida F

キメリズム解析における multiplex short tandem repeat(STR)-PCR 法の検討と検査材料の評価
臨床病理,59(1):24-30 2011(Jan. 25)
Author:平 千明; 松田 和之; 竹澤 由夏; 伊藤 俊朗; 石田 文宏; 日高 惠以子; 熊谷 俊子; 本田 孝行

Quantitative monitoring of single nucleotide mutations by allele-specific quantitative PCR can be used for the assessment of minimal residual disease in patients with hematological malignancies throughout their clinical course
CLINICA CHIMICA ACTA,412(1-2):53-58 2011(Jan.)
Author:Chiaki Taira; Kazuyuki Matsuda; Yuka Kamijyo; Kazuo Sakashita; Fumihiro Ishida; Toshiko Kumagai; Kazuyoshi Yamauchi; Nobuo Okumura; Takayuki Honda
Abstract:Background: Monitoring of minimal residual disease (MRD) in patients with hematological malignancies is important for evaluating the patients' therapeutic response and risk of relapse. Single nucleotide mutations associated with leukemogenesis can be considered as applicable MRD markers. Methods: We developed an allele-specific quantitative polymerase chain reaction (AS-qPCR) for FLT3 2503G>T, KIT2446G>T, and KIT2447A>T and compared the change in the expression levels of the FLT3 or KIT mutations assessed by AS-qPCR to those of the RUNX1-RUNX1T1 fusion gene and WT1 by conventional quantitative PCR. Results: The AS-qPCR using primers including template-mismatched nucleotide or template-mismatched nucleotide plus locked nucleic acid substituted nucleotide provided higher selectivity for mutant nucleotides. The change in the expression levels of the FLT3 or KIT mutations at the time of relapse and just after hematopoietic stem cell transplantation correlated well with that of the RUNX1-RUNX1T1 fusion gene and WT1. Moreover, during complete remission, only AS-qPCR could detect low-level expression of residual mutations. Conclusions: The AS-qPCR for analyzing single nucleotide mutations contributes to the monitoring of MRD in patients without recurrent fusion gene throughout the clinical course and thus broadens the spectrum of patients in whom MRD can be monitored. (C) 2010 Elsevier B.V. All rights reserved.

Donor lymphocyte infusion for extranodal NK/T cell lymphoma, nasal type, relapsed after allogeneic hematopoietic stem cell transplantation
Bone Marrow Transplantation.,46:1270-71 2011
Author:Ito T; Makishima H; Nakazawa H; Senoo Y; Senoo N; Ishida F

Spontaneous regression of the inhibitor against the coagulation factor XIII A subunit in acquired factor XIII deficiency
THROMBOSIS AND HAEMOSTASIS,104(6):1284-1285 2010(Dec.)
Author:Fumihiro Ishida; Kentaro Okubo; Toshiro Ito; Nobuo Okumura; Masayoshi Souri; Akitada Ichinose

Diagnosis and management of natural killer-cell malignancies
EXPERT REVIEW OF HEMATOLOGY,3(5):593-602 2010(Oct.)
Author:Fumihiro Ishida; Yok-Lam Kwong
Abstract:Natural killer (NK)-cell malignancies are uncommon neoplasms, which have been referred to as polymorphic reticulosis or angiocentric T-cell lymphomas in the past. In the current WHO classification, they are categorized as extranodal NK/T-cell lymphoma, nasal type and aggressive NK-cell leukemia. NK-cell malignancies show a geographical predilection for Asian and South American populations and are rare in the west. Pathologically, NK-cell lymphomas show a polymorphic neoplastic infiltrate with angioinvasion and angiodestruction. The lymphoma cells are CD2(+), cytoplasmic CD3 epsilon(+) and CD56(+), with germline T-cell receptor gene. There is an almost invariable clonal episomal infection with Epstein Barr virus. Clinically, NK-cell lymphomas can be classified into nasal, non-nasal and aggressive lymphoma/leukemia subtypes. Most nasal NK-cell lymphomas present with stage I/II disease. The early use of radiotherapy, either alone or concomitantly/sequentially with chemotherapy, is the most important factor in achieving successful treatment. Many stage I/II patients receiving radiotherapy alone fail systemically, so the use of chemotherapy is also considered necessary. Chemotherapy is indicated for stage III/IV nasal NK-cell lymphoma, and the non-nasal and aggressive subtypes. Recent regimens that incorporate the use of L-asparaginase have resulted in substantial improvements in outcome in high-risk, refractory or relapsed patients. High-dose chemotherapy and hematopoietic stem-cell transplantation with autologous or allogeneic hematopoietic stem cells may be beneficial to selected patients. Prognostication of patients with clinical prognostic models and presentation circulating Epstein Barr DNA load may be useful in the stratification of patients for various treatment modalities.

A C-terminal amino acid substitution in the gamma-chain caused by a novel heterozygous frameshift mutation (Fibrinogen Matsumoto VII) results in hypofibrinogenaemia
THROMBOSIS AND HAEMOSTASIS,104(2):213-223 2010(Aug.)
Author:Noriko Fujihara; Ayumi Haneishi; Kazuyoshi Yamauchi; Fumiko Terasawa; Toshiro Ito; Fumihiro Ishida; Nobuo Okumura
Abstract:We found a novel hypofibrinogenemia designated as Matsumoto VII (M-VII), which is caused by a heterozygous nucleotide deletion at position g.7651 in FGG and a subsequent frameshift mutation in codon 387 of the gamma-chain. This frameshift results in 25 amino acid substitutions, late termination of translation with elongation by 15 amino acids, and the introduction of a canonical glycosylation site. Western blot analysis of the patient's plasma fibrinogen visualised with anti-gamma-chain antibody revealed the presence of two extra bands. To identify the extra bands and determine which of the above-mentioned alterations caused the assembly and/or secretion defects in the patient, 11 variant vectors that introduced mutations into the cDNA of the gamma-chain or gamma'-chain were transfected into Chinese hamster ovary cells. In vitro expression of transfectants containing gamma Delta 7651A and gamma Delta 7651A/399T (gamma Delta 7651A with an amino acid substitution of 399Asn by Thr and a variant lacking the canonical glycosylation site) demonstrated a reduction in secretion to approximately 20% of the level seen in the transfectants carrying the normal gamma-chain. Furthermore, results from other transfectants demonstrated that eight aberrant residues between 391 and 398 of the M-VII variant, rather than the 15 amino acid extension or the additional glycosylation, are responsible for the reduced levels of assembly and secretion of M-VII variant fibrinogen. Finally, the results of this study and our previous reports demonstrate that the fibrinogen gamma-chain C-terminal tail (388-411) is not necessary for protein assembly or secretion, but the aberrant amino acid sequence observed in the M-VII variant (especially 391-398) disturbs these functions.

Killer cell immunoglobulin-like receptor gene polymorphism in lymphoproliferative diseases of granular lymphocytes in a Japanese population
LEUKEMIA & LYMPHOMA,51(8):1580-1581 2010(Aug.)
Author:Kosuke Obama; Hideki Makishima; Fumihiro Ishida

Late relapse of extranodal natural killer/T cell lymphoma, nasal type, after more than ten years
LEUKEMIA & LYMPHOMA,51(1):171-173 2010(Jan.)
Author:Fumihiro Ishida; Sayaka Nishina; Naoko Asano; Shigeru Sasaki; Nodoka Sekiguchi; Hideyuki Nakazawa; Toshiro Ito; Naoto Shikama

The diagnosis and management of natural killer cell malignancies.
Expert Review of Hematology,3:593-602 2010
Author:Ishida F, Kwong YL

Age-related Epstein-Barr virus (EBV)-associated B-cell lymphoproliferative disorders: comparison with EBV-positive classic Hodgkin lymphoma in elderly patients
BLOOD,113(12):2629-2636 2009(Mar.)
Author:Naoko Asano; Kazuhito Yamamoto; Jun-Ichi Tamaru; Takashi Oyama; Fumihiro Ishida; Koichi Ohshima; Tadashi Yoshino; Naoya Nakamura; Shigeo Mori; Osamu Yoshie; Yoshie Shimoyama; Yasuo Morishima; Tomohiro Kinoshita; Shigeo Nakamura
Abstract:Age-related Epstein-Barr virus-associated B-cell lymphoproliferative disorder (aEBVLPD) is a disease group characterized by EBV-associated large B-cell lymphoma in the elderly without predisposing immunodeficiency. In nearly one-third of cases, aEBVLPD occurs as a polymorphous subtype with reactive cell-rich components, bearing a morphologic similarity to classic Hodgkin lymphoma (cHL). The aim of this study was to clarify clinicopathologic differences between the polymorphic subtype of aEBVLPD (n = 34) and EBV+ cHL (n = 108) in patients aged 50 years or older. Results showed that aEBVLPD was more closely associated with aggressive clinical parameters than cHL, with a higher age at onset (71 vs 63 years); lower male predominance (male-female ratio, 1.4 vs 3.3); and a higher rate of involvement of the skin (18% vs 2%), gastrointestinal tract (15% vs 4%), and lung (12% vs 2%). aEBVLPD was histopathologically characterized by a higher ratio of geographic necrosis, greater increase (>30%) in cytotoxic T cells among background lymphocytes, higher positivity for CD20 and EBNA2, and absence of CD15 expression. As predicted by the clinical profile, aEBVLPD had a significantly poorer prognosis than EBV+ cHL (P <.001). The polymorphous subtype of aEBVLPD constitutes an aggressive group with an immune response distinct from EBV+ cHL, and requires the development of innovative therapeutic strategies. (Blood. 2009; 113: 2629-2636)

Promising approach for aggressive NK cell leukaemia with allogeneic haematopoietic cell transplantation
Author:Toshiro Ito; Hideki Makishima; Hideyuki Nakazawa; Hikaru Kobayashi; Shigetaka Shimodaira; Yozo Nakazawa; Kiyoshi Kitano; Kazuyuki Matsuda; Eiko Hidaka; Fumihiro Ishida
Abstract:Objectives: Aggressive natural killer cell leukaemia (ANKL) is a malignant disorder of mature NK cells with a poor prognosis, for which no effective therapeutic approach has been established. We investigated the role of allogeneic haematopoietic cell transplantion (allo-HCT) in ANKL. Patients and methods: Three patients with ANKL received allo-HCT and seven did not. Epstein-Barr virus (EBV) viral load (VL) of the whole blood was measured with real-time quantitative polymerase chain reaction. Results: We transplanted three patients using a myeloablative conditioning regimen with human leucocyte antigen (HLA) two-loci mismatched cord blood (n=2), or HLA-matched sibling bone marrow (n=1). In one patient, a second transplantation from the haploidentical mother was also performed at relapse. No patients were in complete remission (CR) at the time of conditioning. After allo-HCT, all three achieved and maintained CR. One died from sepsis and the other relapsed, received the second transplantation and achieved a second CR. EBV VL was quite high in all three at presentation and its significant reduction was observed after allo-HCT. Although their backgrounds were not different from those without allo-HCT, patients with allo-HCT had a better outcome. Conclusion: Allo-HCT might be a promising therapy for ANKL with curative potential.

Additional chromosomal changes in impaired chimeric analysis of a patient with relapsed leukemia after bone marrow transplantation
臨床血液,49 2008(Feb.)
Author:Higuchi Y; Ito T; Matsuda K; Higuchi T; Hidaka E; Imagawa E; Uhara M; Nakazawa H; Ishida F; Yamauchi K; Sano K; Katsuyama T

Zygomycosis presenting as acute myocardial infarction during hematological malignancies
INTERNAL MEDICINE,47(9):839-842 2008
Author:Satoru Joshita; Kiyoshi Kitano; Tadanobu Nagaya; Atsushi Kamijo; Koh Nakazawa; Fumihiro Ishida
Abstract:Here we report two patients with hematological malignancies associated with complications of fatal cardiac zygomycosis. The first case, a 72-year-old man with myelodysplastic syndrome being treated with low-dose cytarabine, died of sudden cardiac arrest. An autopsy revealed disseminated zygomycosis accompanied with occlusion of the coronary artery by fungal thrombi. The second case, a 52-year-old woman with acute lymphoblastic leukemia, developed febrile neutropenia and skin eruptions with induration on the face and extremities during the first induction chemotherapy. She experienced sudden bradycardia with unstable hemodynamics and died of acute myocardial infarction. Histological examination of a skin biopsy demonstrated zygomycosis. In light of the above, it should be kept in mind that cardiac zygomycosis might occur in hematologically compromised patients presenting with acute myocardial infarction.

Peripheral T-cell lymphoma following diffuse large B-cell lymphoma associated with celiac disease.
Internal medicine (Tokyo, Japan),47(4):295-298 2008
Author:Makishima H; Komiyama Y; Asano N; Momose K; Nakamura S; Ishida F

Chemokine system and tissue infiltration in aggressive NK-cell leukemia
LEUKEMIA RESEARCH,31(9):1237-1245 2007(Sep.)
Author:Hideki Makishima; Toshiro Ito; Kayoko Momose; Hideyuki Nakazawa; Shigetaka Shimodaira; Yuji Kamijo; Yozo Nakazawa; Naoaki Ichikawa; Mayumi Ueno; Hikaru Kobayashi; Kiyoshi Kitano; Hiroshi Saito; Kendo Kiyosawa; Fumihiro Ishida
Abstract:NK cell-type lymphoproliferative disease of granular lymphocytes can be subdivided into aggressive NK-cell leukemia (ANKL) and chronic NK-cell lymphocytosis (CNKL). Hepatosplenomegaly is observed in ANKL patients, and hepatic failure is a common cause of death. Significant numbers of ANKL cells were pathologically observed in sinusoidal and interlobular regions of the liver, and in the splenic red pulp. In our previous study, ANKL cells were simultaneously positive for CXCR1 and CCR5. So, in order to elucidate the mechanism in the systemic migration of ANKL cells, we investigated the expression of the corresponding chemokines in ANKL compared with CNKL. The serum level of IL-8, MIP-1 alpha and MIP-1 beta was significantly elevated in ANKL patients, and ANKL cells were highly positive for IL-8, RANTES, MIP-1 alpha and MIP-1 beta according to intracellular staining and RT-PCR. These chemokines were also positively stained in heparocytes. The interaction between Fas and Fas ligand (FasL) is supposed to be one of the mechanisms for liver dysfunction in ANKL. The serum concentration of soluble FasL was significantly high in ANKL patients, and ANKL cells expressed FasL protein in the cytoplasm. These results suggest that the chemokine system plays an important role in the transmigration of FasL-expressing ANKL cells. (c) 2006 Elsevier Ltd. All rights reserved.

Close resemblance between chemokine receptor expression profiles of lymphoproliferative disease of granular lymphocytes and their normal counterparts in association with elevated serum concentrations of IP-10 and MIG.
International journal of hematology,86(2):174-179 2007(Aug.)
Author:Momose K; Makishima H; Ito T; Nakazawa H; Shimodaira S; Kiyosawa K; Ishida F

Cytotoxic molecule expression is predictive of prognosis in Hodgkin's-like anaplastic large cell lymphoma
Histopathology,50(6):705-715 2007
Author:Asano N; Suzuki R; Matsuo K; Kagami Y; Ishida F; Tamaru J.-I; Jin G.-S; Sato Y; Shimoyama Y; Yoshino T; Morishima Y; Nakamura S

A case of acute myelogenous leukemia with MLL-AF10 fusion caused by insertion of 5 ' MLL into 10p12, with concurrent 3 ' MLL deletion
Author:Kazuyuki Matsuda; Eiko Hidaka; Futnihiro Ishida; Kazuyoshi Yamauchi; Hideki Makishima; Toshiro Ito; Takefumi Suzuki; Eri Imagawa; Kenji Sano; Tsutomu Katsuyama; Hiroyoshi Ota
Abstract:Structural abnormalities involving the mixed-lineage leukemia (MLL) gene on 11q23 have been associated with hematological malignancies. The rearrangement of MLL occurs during translocations and insertions involving a variety of genes on the partner chromosome. We report a rare case of acute myelogenous leukemia (AML-M2) with 11q23 abnormalities. Fluorescence in situ hybridization (FISH) using a commercial dual-color MLL probe detected an atypical signal pattern: one fusion signal, two green signals smaller than those usually detected, and no orange signals. Spectral karyotyping (SKY) analysis indicated that one green signal was detected on the short arm of derivative chromosome 10, and the other green signal on the long arm of a derivative chromosome 11, on which no orange signal was detected. A long-distance inverse polymerase chain reaction (LDI-PCR) identified the fusion partner gene, in which introm 6 of MLL was fused with intron 8 of AF10 on 10p12 in the 5' to 3' direction. Our observations indicated that the MLL-AF10 fusion gene resulted from the insertion of part of the region that included the 5' MLL insertion into 10p12; this was concurrent with the deletion of 3' MLL. (c) 2006 Elsevier Inc. All rights reserved.

Prognostic significance of T-cell or cytotoxic molecules phenotype in classical Hodgkin's lymphoma: A clinicopathologic study
JOURNAL OF CLINICAL ONCOLOGY,24(28):4626-4633 2006(Oct.)
Author:Naoko Asano; Aya Oshiro; Keitaro Matsuo; Yoshitoyo Kagami; Fumihiro Ishida; Ritsuro Suzuki; Tomohiro Kinoshita; Yoshie Shimoyama; Jun-Ichi Tamaru; Tadashi Yoshino; Kunio Kitamura; Hisashi Fukutani; Yasuo Morishima; Shigeo Nakamura
Abstract:Purpose Classical Hodgkin's lymphoma (CHL) is characterized by Hodgkin's and Reed-Sternberg (H-RS) cells, most of which are derived from germinal-center B cells. Nevertheless, one or more markers for T cells and follicular dendritic cells (FDC) may be expressed in a minority of H-RS cells in some CHL patients, although the clinical significance of this remains controversial. The aim of this study was to clarify the association between phenotypic expression and clinical outcome in CHL. Patients and Methods Participants were 324 consecutive CHL patients, comprising 132 patients with nodular sclerosis (NS), 35 patients with NS grade 2 (NS2), and 157 patients with mixed cellularity (MC). We evaluated the presenting features and prognosis of patients on categorization into four phenotypically defined groups: B-cell (CD20(+) and/or CD79a(+); n = 63), T-cell and/or cytotoxic molecules (CD3(+), CD4(+), CD8(+), CD45RO(+), TIA-1(+), and/or granzyme B+; n = 27), FDC (CD21(+) without B-cell marker, n = 22), and null-cell types (n 212). Other potential prognostic factors were examined. Results The T-cell and/or cytotoxic molecules group showed a significantly poorer prognosis than the other three groups (P <.0001). This finding was seen consistently in multivariate analyses. Morphologic subtyping (NS/NS2/MC) and Epstein-Barr virus positivity were not identified as independent prognostic factors. Conclusion The presence of T-cell and/or cytotoxic antigens in H-RS cells may represent a poor prognostic factor in CHL, even if their expression is not regarded as lineage specific. Examination of T-cell and/or cytotoxic molecules phenotype in CHL patients is recommended as a routine pathologic practice.

Intestinal diffuse large B-cell lymphoma associated with celiac disease: A Japanese case
Author:H Makishima; T Ito; R Kodama; N Asano; H Nakazawa; K Hirabayashi; S Nakamura; M Ota; T Akamatsu; K Kiyosawa; F Ishida
Abstract:Intestinal non-Hodgkin's lymphoma (NHL), especially the T-cell type, is well known to be associated with celiac disease (CD), an enteropathic disorder with a propensity for certain racial and genetic backgrounds. CD is typically characterized by gastrointestinal (GI) symptoms, anti-transglutaminase antibodies in the sera, and microscopical findings of the intestinal mucosa, which resolve with a gluten-free diet (GFD). In Asian populations, including the Japanese, CD and the associated NHL have been supposed to be quite rare, and studies concerning the frequency of CD or its relationship with NHL are scarce. We describe a Japanese middle-aged man with intestinal diffuse large B-cell lymphoma associated with CD. Following multi-combined chemotherapy, the patient's lymphoma has been in a state of complete response, and his GI symptoms have improved with a GFD. This case suggests that the possibility of CD and its association with intestinal NHL should be kept in mind, even in Asian populations.

Adenovirus fulminant hepatic failure: Disseminated adenovirus disease after unrelated allogeneic stem cell transplantation for acute lymphoblastic leukemia
INTERNAL MEDICINE,45(16):975-980 2006
Author:Hideyuki Nakazawa; Toshiro Ito; Hideki Makishima; Noriko Misawa; Wataru Okiyama; Takeshi Uehara; Eiko Hidaka; Kendo Kiyosawa; Fumihiro Ishida
Abstract:Adenovirus is one of the major causes of non-relapse morbidity and mortality after allogeneic hematopoietic stem cell transplantation for hematological malignancy. Fulminant hepatic failure is a rare manifestation of post-transplant complication with adenovirus. Extremely high mortality and aggressiveness of the clinical course have been posing clinical challenges for the diagnosis as well as for the treatment. Here, we report a case with disseminated adenovirus disease presenting with fulminant hepatic failure after bone marrow transplantation for acute lymphoblastic leukemia.

Thrombotic thrombocytopenic purpura associated with pegylated-interferon alpha-2a by an ADAMTS13 inhibitor in a patient with chronic hepatitis C
Haematologica,91:ECR34 2006
Author:Kitano K; Gibo Y; Kamijo A; Furuta K; Oguchi S; Joshita S; Takahashi Y; Ishida F; Matsumoto M; Uemura M; Fujimura Y

Prognostic significance of T-cell or cytotoxic molecules phenotype in classical Hodgkin’s lymphoma: a clinicopathologic study
J Clin Oncol,24:2646-2633 2006
Author:Asano N; Oshiro A; Matsuo K; Kagami Y; Ishida F; Suzuki R; Kinoshita T; Shimoyama Y; Tamaru J; Yoshino T; Kitamura K; Fukutani H; Morishima Y; Nakamura S

Clinicopathologic and prognostic significance of cytotoxic molecule expression in nodal peripheral T-cell lymphoma, unspecified
Author:N Asano; R Suzuki; Y Kagami; F Ishida; K Kitamura; H Fukutani; Y Morishima; K Takeuchi; S Nakamura
Abstract:Cytotoxic molecules (CMs) are apoptosis-inducing molecules that are present in azurophilic cytoplasmic granules of T lymphocytes. Expression of TIA-1 and granzyme B was examined for 100 cases of nodal peripheral T-cell lymphoma, unspecified (PTCL-U) to assess clinicopathologic significance of CM. Forty-one were positive for at least one CM. Patients with CM-positive PTCL-U showed younger onset (median, 55 years vs. 64 years, P = 0.01) and less male predominance (male:female ratio, 21:20 vs. 44:15, P = 0.02). CM-positive PTCL-U was significantly associated with several clinical factors to indicate poor prognosis, in comparison with CM-negative PTCL-U, such as poorer performance status (P = 0.006), more frequent B-symptoms (68% vs. 35%, P = 0.002), higher serum lactate dehydrogenase levels (P = 0.003), and more frequent extranodal involvement, particularly bone marrow involvement (33% vs. 9%, P = 0.004). Epstein-Barr virus was mostly found in CM-positive PTCL-U (51%vs. 2%, P < 0.0001). The CM-positive group showed higher distribution of the International Prognostic Index (P = 0.009) and the Prognostic Index for T-cell lymphoma (P = 0.004) scores than CM-negative group. Complete remission rate was 30% for the former but 63% for the latter. Overall survival of CM-positive PTCL-U was significantly lower than that of CM-negative patients (P = 0.004). Multivariate analyses confirmed at CM expression is a significant prognostic factor, independent from of er clinical factors or prognostic index scores. These findings suggest that nodal CM-positive PTCL-U show distinct clinicopathologic characteristics among the current category of PTCL-U.

Significance of chemokine receptor expression in aggressive NK cell leukemia
LEUKEMIA,19(7):1169-1174 2005(Jul.)
Author:H Makishima; T Ito; N Asano; H Nakazawa; S Shimodaira; Y Kamijo; Y Nakazawa; T Suzuki; H Kobayashi; K Kiyosawa; F Ishida
Abstract:Natural killer (NK) cell-type lymphoproliferative diseases of granular lymphocytes can be subdivided into aggressive NK cell leukemia (ANKL) and chronic NK cell lymphocytosis (CNKL). One reason for the poor outcome in ANKL is leukemic infiltration into multiple organs. The mechanisms of cell trafficking associated with the chemokine system have been investigated in NK cells. To clarify the mechanism of systemic migration of leukemic NK cells, we enrolled nine ANKL and six CNKL cases, and analyzed the expression profiles and functions of chemokine receptors by flowcytometry and chemotaxis assay. CXCR1 was detected on NK cells in all groups, and CCR5 was positive in all ANKL cells. Proliferating NK cells were simultaneously positive for CXCR1 and CCR5 in all ANKL patients examined, and NK cells with this phenotype did not expand in CNKL patients or healthy donors. ANKL cells showed enhanced chemotaxis toward the ligands of these receptors. These results indicated that the chemokine system might play an important role in the pathophysiology of ANKL and that chemokine receptor profiling might be a novel tool for discriminating ANKL cells from benign NK cells.

DNA microarray analysis of natural killer cell-type lymphoproliferative disease of granular lymphocytes with purified CD3(-)CD56(+) fractions
LEUKEMIA,18(3):556-565 2004(Mar.)
Author:YL Choi; H Makishima; J Ohashi; Y Yamashita; R Ohki; K Koinuma; J Ota; Y Isobe; F Ishida; K Oshimi; H Mano
Abstract:Natural killer (NK) cell-type lymphoproliferative disease of granular lymphocytes (LDGL) is characterized by the outgrowth of CD3(-)CD16/56(+) NK cells, and can be further subdivided into two distinct categories: aggressive NK cell leukemia (ANKL) and chronic NK lymphocytosis (CNKL). To gain insights into the pathophysiology of NK cell-type LDGL, we here purified CD3(-)CD56(+) fractions from healthy individuals (n=9) and those with CNKL (n=9) or ANKL (n=1), and compared the expression profiles of >12000 genes. A total of 15 'LDGL-associated genes' were identified, and a correspondence analysis on such genes could clearly indicate that LDGL samples share a 'molecular signature' distinct from that of normal NK cells. With a newly invented class prediction algorithm, 'weighted distance method', all 19 samples received a clinically matched diagnosis, and, furthermore, a detailed cross-validation trial for the prediction of normal or CNKL status could achieve a high accuracy (77.8%). By applying another statistical approach, we could extract other sets of genes, expression of which was specific to either normal or LDGL NK cells. Together with sophisticated statistical methods, gene expression profiling of a background-matched NK cell fraction thus provides us a wealth of information for the LDGL condition.

Ribavirin-induced pure red-cell aplasia during treatment of chronic hepatitis C.
NEW ENGLAND JOURNAL OF MEDICINE,350(12):1264-1265 2004(Mar.)
Author:N Tanaka; F Ishida; E Tanaka

Idiopathic myelofibrosis with refractory massive ascites
INTERNAL MEDICINE,42(6):525-528 2003(Jun.)
Author:M Yotsumoto; F Ishida; T Ito; M Ueno; K Kitano; K Kiyosawa
Abstract:A 55-year-old woman presented with massive refractory ascites in the course of idiopathic myelofibrosis. The ascites was exudative, and a cytological examination revealed granulocytes of varying maturity, erythroblasts, and megakaryocytes with trisomy 8. The ascites was assumed to have developed from peritoneal extramedullary hematopoiesis. An abnormal karyotype in the cells in the ascitic fluid, which was the same abnormality as in peripheral blood, helped to prove extramedulary hematopoiesis in this case, which can be an aid in making a differential diagnosis in cases of ascites associated with myelofibrosis.

Lymphoproliferative disease of granular lymphocytes with T-cell receptor gamma delta-positive phenotype: restricted usage of T-cell receptor gamma and delta subunit genes
Author:H Makishima; F Ishida; H Saito; N Ichikawa; Y Ozaki; S Ito; M Ota; Y Katsuyama; K Kiyosawa
Abstract:Lymphoproliferative disease of granular lymphocytes (LDGL) is characterized by more than 0.5 x 10(9)/L of proliferating granular lymphocytes in the peripheral blood. Because of its rarity, the characteristics of LDGL with T-cell receptor (TCR) gammadelta phenotype (gammadelta T-LDGL) have not yet been identified. This report describes the clinical, hematological, and immunological findings of four patients with this disease. In two cases, the clinical course was indolent and the other two patients required various therapies. The cells had a common immunophenotype: CD3+, CD4-, CD16+, CD56-, CD57-, CD122-, TCR-gammadelta+, and three were CD8-positive. The immunopurified TCR-gammadelta cells from the patients expressed only Vgamma9 and Vdelta1. Spectratyping and sequencing showed mono- or oligoclonality for TCRgamma and TCRdelta subunit genes. Soluble Fas ligand in sera was significantly elevated in all patients. These findings suggest that gammadelta T-LDGL qualifies as a distinct disease entity.

DNA microarray analysis of T cell-type lymphoproliferative disease of granular lymphocytes
BRITISH JOURNAL OF HAEMATOLOGY,118(2):462-469 2002(Aug.)
Author:H Makishima; F Ishida; T Ito; K Kitano; S Ueno; K Ohmine; Y Yamashita; J Ota; M Ota; K Yamauchi; H Mano
Abstract:Lymphoproliferative disease of granular lympho- cytes (LDGL) is characterized by the clonal proliferationoflarge granular lymphocytes of either T- or natural killer cell origin. To better understand the nature of T cell-type LDGL, we purified the CD4(-) CD8(+) proliferative fractions from LDGL patients (n =4) and the surface marker-matched T cells isolated from healthy volunteers (n =4), and compared the expression profiles of 3456 genes using DNA microarray. Through this analysis, we identified a total of six genes whose expression was active in the LDGL T cells, but silent in the normal ones. Interestingly, expression of the gene for interleukin (IL) 1beta was specific to LDGL T cells, which was further confirmed by the examination of the serum level of IL-1beta protein. Given its important role in inflammatory reactions, the disease-specific expression of IL-1beta may have a causative relationship with the LDGL- associated rheumatoidarthritis. Spectratyping analysis of the T-cell receptor repertoire also proved the monoclonal or oligoclonal natureof LDGL cells. These data have shown that microarray analysis with a purified T-cell subset is an efficient approach to investigate the pathological condition of Tcell-type LDGL.

Human factor VII deficiency caused by S339C mutation located adjacent to the specificity pocket of the catalytic domain
Author:O Takamiya; M Seta; K Tanaka; F Ishida
Abstract:This report documents our identification of a novel factor VII (FVII) gene mutation in a Japanese boy with FVII deficiency. The proband's FVII activity was 34% and his FVII antigen level was 40% of normal controls. DNA sequence analysis of the proband's FVII gene identified a C to G point mutation at nucleotide position 10 933 in exon 8, which results in the substitution of Cys (TGC) for Ser339 (TCC). Hinf I digestion results indicate the proband and his mother were heterozygous for the mutation. Both wild-type and mutant FVIIs were transiently expressed in COS-1 cells. FVII levels measured in the culture medium of FVII Ser339Cys mutants were markedly reduced as compared to those of cells with FVII wild-type. The amount of intracellular FVII in FVII Ser339Cys mutants was 80% of that in wild-type. In the wild-type FVII, Ser339 is juxtaposed to Asp338, which is positioned at the bottom of the substrate-binding pocket in the protease domain and located adjacent to FVII Cys340, that forms a disulphide bond with Cys368. We suspect that the creation of a novel unpaired cysteine through this mutation leads to abnormal disulphide bonding during protein folding, thereby reducing the secretion of FVII.

t(8;21;14)(q22;q22;q24) is a novel variant of t(8;21) with chimeric transcripts of AML1-ETO in acute myelogenous leukemia
Author:F Ishida; M Ueno; H Tanaka; H Makishima; K Suzawa; S Hosaka; E Hidaka; M Ishikawa; K Yamauchi; K Kitano; K Kiyosawa
Abstract:We report a male patient with acute myelogenous leukemia (AML; French-American-British M2) associated with AML 1-ETO. Cytogenetic studies showed a complex karyotype including a novel translocation (8;21;14)(q22;q22;q24) in all analyzed cells. This three-way translocation was confirmed with spectral karyotyping. Reverse transcription-polymerase chain reaction analysis for AML1-ETO chimeric transcripts showed the presence of the fusion product with the expected size. Translocation (8;21;14)(q22; q22;q24) is a novel variant of t(8;21)(q22;q22), possibly having a common molecular pathogenetic mechanism. (C) 2002 Elsevier Science Inc. All rights reserved.

A missense mutation (Tyr88 to Cys) in the platelet membrane glycoprotein Ib beta gene affects GPIb/IX complex expression - Bernard-Soulier syndrome in the homozygous form and giant platelets in the heterozygous form
THROMBOSIS AND HAEMOSTASIS,86(5):1249-1256 2001(Nov.)
Author:Y Kurokawa; F Ishida; T Kamijo; S Kunishima; D Kenny; K Kitano; K Koike
Abstract:This study examined the Molecular basis of a missense mutation of the platelet glycoprotein (GP) Ib beta gene in two families. In the propositus with a novel form of Bernard-Soulier syndrome (BSS) from Family I, only GPIb alpha was detectable in reduced amounts on platelet surfaces by flow cytometry. There were no GPIX or GPIb beta found by immunoblotting. DNA sequencing analysis showed a homozygous mutation in the GPIb beta gene which changed Tyr (TAC) to Cys (TGC) at residue 88. Her parents were heterozygous for Tyr88Cys in the GPIb beta gene. In transient transfection studies on 293T cells, both Tyr88Cys and Tyr88Ala mutations suppressed the expression of GPIb/IX complexes. in addition, Tyr88Cys GPIb beta mutation was found to exert a dominant negative effect on the GPIb alpha expression. Five individuals from Family II, four of whom reported elsewhere as having giant platelet disorders with normal aggregation (BLOOD, 1997 89: 2404) and one newly analyzed in this study, were heterozygous for Tyr88Cys in the GPIb beta gene. Microsatellite analysis of chromosome 22 showed a common haplotype in 8 of the individuals with Tyr88Cys mutations in Families I and II. Tyr88 in the GPIb beta gene plays a significant role in the GPIb/IX expression; the defect causes BSS in a homozygous form and possibly giant platelets in a heterozygous form.

Fibrinogen Matsumoto V: a variant with a alpha 19 Arg -> Gly (AGG -> GGG) - Comparison between fibrin polymerization stimulated by thrombin or reptilase and fibrin monomer polymerization
THROMBOSIS AND HAEMOSTASIS,85(1):108-113 2001(Jan.)
Author:H Tanaka; F Terasawa; T Ito; S Tokunaga; F Ishida; K Kitano; K Kiyosawa; N Okumura
Abstract:Fibrinogen Matsumoto V (M-V) is a dysfibrinogen identified in a 52-year-old woman with systemic lupus erythematous. The triplet AGG encoding the amino acid residue A alpha 19 was replaced by GGG, resulting in the substitution of Arg-->Gly. Residue Aa19 has been shown to be one of the most important amino acids in the so-called 'A' site or a-chain knob. The thrombin-catalyzed release of fibrinopeptide A from M-V fibrinogen was only slightly delayed yet release of fibrinopeptide B was significantly delayed. Both thrombin-catalyzed fibrin polymerization and fibrin monomer polymerization were markedly impaired compared to normal fibrinogen. In addition, reptilase-catalyzed fibrin polymerization of M-V was much more impaired than thrombin-catalyzed fibrin polymerization. These results indicate 'B' and/or 'b' site of M-V fibrinogen play a more important role in thrombin-catalyzed fibrin polymerization than that of normal control fibrinogen.

Hairy cell leukemia with translocation (11;20)(q13;q11) and overexpression of cyclin D1
LEUKEMIA RESEARCH,23(8):763-765 1999(Aug.)
Author:F Ishida; K Kitano; N Ichikawa; T Ito; Y Kohara; T Taniguchi; T Motokura; K Kiyosawa
Abstract:We report on a male Japanese patient with hairy cell leukemia (HCL). A cytogenetic study with lipopolysaccharide stimuli showed a novel translocation (11;20)(q13;q11) in 10% of the analyzed cells. Northern blot analysis and RT-PCR analysis for cyclin D1 revealed the overexpression of cyclin D1, although the southern blot analysis of PRAD1 gene showed no rearrangement. In this particular case, the t(11;20)(q13: q11) might play some role in the oncogenesis of HCL and the overexpression of cyclin D1. (C) 1999 Elsevier Science Ltd. All rights reserved.

Fibrinogen Matsumoto III: a variant with gamma 275 Arg -> Cys (CGC -> TGC) - Comparison of fibrin polymerization properties with those of Matsumoto I (gamma 364 Asp -> His) and Matsumoto II (gamma 308 Asn -> Lys)
THROMBOSIS AND HAEMOSTASIS,81(5):763-766 1999(May)
Author:F Terasawa; N Okumura; Y Higuchi; S Ishikawa; M Tozuka; F Ishida; K Kitano; T Katsuyama
Abstract:Fibrinogen Matsumoto III (M-III) is a dysfibrinogen identified in a 66-year-old woman with rectal cancer. The fibrinogen level determined by the thrombin-time method was markedly decreased in preoperative coagulation tests of her plasma. Three fibrinogen polypeptide-chain gene fragments from the proposita were amplified by the polymerase chain reaction method, then sequenced. The triplet CGC encoding the amino acid residue gamma 275 was replaced by TGC, resulting in the substitution of Arg-->Cys. There have been previous reports of nine families with the same alteration, nine families with an Arg-->His variant and one family with an Arg-->Ser variant in this residue, which has been shown to be one of the most important amino acids in the 'D:D' interaction site. In addition, there are three silent mutations in the A alpha-chain gene and two mutations in the intron of the B beta-chain and the gamma-chain gene. However, none of these mutations is thought to be the cause of the dysfunctional fibrinogen. The thrombin-catalyzed fibrin polymerization in the presence of 1 mM Ca ions was markedly delayed in purified M-III. Its lag period was longer than those of Matsumoto II (M-II; gamma 308Asn-->Lys) and Matsumoto 1 (M-I; gamma 364Asp-->His). gamma 364Asp is one of the most important residues in the polymerization pocket of the 'D:E' interaction site and gamma 308Asn is located in the vicinity of a high affinity Ca(2+) binding site in the D-domain, gamma 311-336. The maximum slope of the polymerization curve for M-III was about 4-fold steeper than that for M-I but less steep than that for M-II. These results may suggest that the tertiary structure of the polymerization pocket plays a more important role in the lateral aggregation of protofibrils than that of the 'D:D' interaction site.

Abnormal proliferation of CD4(-) CD8(+) gamma delta(+) T cells with chromosome 6 anomaly: Role of fas ligand expression in spontaneous regression of the cells
Author:N Ichikawa; K Kitano; T Ito; T Nakazawa; S Shimodaira; F Ishida; K Kiyosawa
Abstract:We report a case of granular lymphocyte proliferative disorder accompanied with hemolytic anemia and neutropenia. Phenotypes of the cells were T cell receptor gamma delta(+) CD3(+) CD4(-)CD8(+) CD16(+) CD56(-) CD57(-), Southern blot analysis of T cell receptor beta and gamma chains demonstrated rearranged bands in both. Chromosomal analysis after IL-2 stimulation showed deletion of chromosome 6, Sorted gamma delta(+) T cells showed an increase in Pas ligand expression compared with the levels in sorted alpha beta(+) T cells, The expression of Pas ligand on these gamma delta(+) T cells increased after IL-2 stimulation, The patient's anemia improved along with a decrease in granular lymphocyte count and disappearance of the abnormal karyotype without treatment. The expression of Pas ligand may be involved in spontaneous regression of granular lymphocyte proliferation with hemolytic anemia, (C) 1999 Wiley-Liss, Inc.

Elliptocytosis in myelodysplastic syndrome associated with translocation (1;5)(p10;q10) and deletion of 20q
Author:F Ishida; S Shimodaira; H Kobayashi; H Saito; M Kako; A Kanzaki; Y Yawata; K Kitano; K Kiyosawa
Abstract:Acquired elliptocytosis is a red blood cell abnormality occasionally associated with myelodysplastic syndrome (MDS). A Japanese male with MDS who presented with elliptocytosis had mild anemia and hypercellular bone marrow with three lineage-dysplasia. He was diagnosed with refractory anemia of MDS. Cytogenetic analysis of bone marrow cells showed 47,XY, +1,der(1;5)(q10;p10),t(1;5) (p10;q10),del(20)(q11) in 70% of the analyzed cells. Analysis of red blood cell membrane proteins by sodium dodecyl sulphate-polyacrylamide gel electrophoresis showed normal electrophoretic patterns with no quantitative abnormalities of each protein. Del(20q) and/or t(1;5)(p10;q10) might be associated with elliptocytosis in this patient. (C) Elsevier Science Inc., 1998.

Liver cirrhosis with marked thrombocytopenia and highly elevated serum thrombopoietin levels
International Journal of Hematology,70(1):52-55 1999
Author:Kiyoshi Kitano; Shigetaka Shimodaira; Toshiro Ito; Naoaki Ichikawa; Hiroshi Kodaira; Yoichi Kohara; Mayumi Ueno; Tomoyuki Tahara; Takashi Kato; Fumihiro Ishida; Kendo Kiyosawa
Abstract:Three patients with liver cirrhosis (LC) and a bleeding tendency due to marked thrombocytopenia of less than 20 × 109/1 were admitted to our hospital for further examination. Bone marrow examination revealed megakaryocytic hypoplasia in all three patients. All patients exhibited amegakaryocytic thrombocytopenic purpura, myelodysplastic syndrome, or bone marrow hypoplasia. 111In-labeled platelet kinetic studies revealed decreased platelet production in all patients. Although serum thrombopoietin (sTPO) levels are usually within the normal level in patients with LC, the sTPO levels of our patients were about 10 times higher than the levels of normal subjects (1.22 ± 0.37 fmol/ml): 13.34, 16.79, and 10.46 fmol/ml, respectively. These sTPO data supported our findings of decreased megakaryopoiesis. Our findings suggest that examination of sTPO lev els is useful in determining the etiology of marked thrombocytopenia in LC patients. © 1999 The Japanese Society of Hematology.

Polymorphisms of platelet membrane glycoprotein Ibα and plasma von Willebrand factor antigen in coronary artery disease
International Journal of Hematology,70(1):47-51 1999
Author:Toshiro Ito; Fumihiro Ishida; Shigetaka Shimodaira; Kiyoshi Kitano
Abstract:To examine the relationships of two polymorphisms of platelet glycoprotein (GP) Ibα and coronary artery diseases (CAD) in Japanese patients, we conducted a case-control study with 158 Japanese patients and 169 control subjects. The frequencies of HPA-2 polymorphism and the variable number of tandem repeat (VNTR) polymorphisms in the macroglycopeptide region did not significantly differ between CAD patients and control subjects. The polymorphisms of GPIbα were not associated with the number of affected vessels in CAD patients. When patients with acute coronary syndrome only were analyzed, the frequencies of the two polymorphisms of GPIbα showed no significant difference. Although plasma von Willebrand antigen (vWF:Ag) levels in patients were significantly higher than in controls, no association between vWF concentration and GPIb genotypes was observed. In patient groups with higher or lower vWF:Ag concentrations, no increase in the frequencies of Met145 or larger VNTR polymorphisms was seen in either group. Our findings indicate that no association exists between the frequencies of the two polymorphisms of GPIbα and CAD. © 1999 The Japanese Society of Hematology.

Associations between polymorphism of platelet membrane glycoprotein Ib alpha, plasma von Willebrand factor antigen and coronary artery disease.
BLOOD,92(10):191A-191A 1998(Nov.)
Author:T Ito; F Ishida; S Shimodaira; M Takei; K Kitano; K Kiyosawa

Cold agglutinin disease by autoanti-i blood type antibody associated with B cell chronic lymphocytic leukemia
Author:F Ishida; H Saito; K Kitano; K Kiyosawa
Abstract:Cold agglutinin (CA) disease is a rare complication of B cell chronic lymphocytic leukemia (CLL). We report a case of CA disease by autoanti-i blood type antibody associated with CDS-negative CLL. The specificity of cold agglutinin in the sera and eluates from red blood cells were analyzed by reactivity against panels of red blood cells and also hemagglutinin inhibition assay using lactonorhexaosylceramide, and were determined to be an anti-i blood group antibody of the monoclonal IgM kappa isotype. PCR analysis of immunoglobulin heavy-chain genes from bone marrow-derived genomic DNA showed CLL cells utilizing DP-54 as a VH gene, differing from the reported VH genes used for cold agglutinin with anti-I/i specificity. The anti-i antibody in this patient might be secondarily produced by residual B cells due to an immunodysregulatory state, and not from CLL cells. (C) 1998 Elsevier Science Ireland Ltd. All rights reserved.

Changes in serum thrombopoietin levels after splenectomy
Acta Haematologica,100(3):137-141 1998
Author:Naoaki Ichikawa; Kiyoshi Kitano; Shigetaka Shimodaira; Fumihiro Ishida; Toshiro Ito; Shoji Kajikawa; Tomoyuki Tahara; Takashi Kato; Kendo Kiyosawa
Abstract:To clarify the role of thrombopoietin (c-Mp1 ligand, TPO) in 'hypersplenic' thrombocytopenia, we used an enzyme-linked immunosorbent assay to examine changes in serum TPO levels accompanied with splenectomy in 6 patients with liver cirrhosis, 4 patients with gastric cancer, and 2 patients with lymphoid malignancies. We also measured serum levels of other thrombopoietic cytokines such as interleukin-6 (IL-6) and erythropoietin. Platelet counts reached a maximum at day 14 after splenectomy in all subjects. In patients with liver cirrhosis, a lower elevation of platelet counts was observed compared with that in patients with gastric cancer. Serum TPO levels gradually elevated after splenectomy and reached a maximum 3.5 days after splenectomy in noncirrhotic patients, whereas peak serum TPO levels were delayed until day 7 in the cirrhosis group. IL-6 and erythropoietin showed similar kinetics between cirrhotic and noncirrhotic patients. These findings suggest that transient thrombocytosis after splenectomy may be associated with an alteration in the site of TPO catabolism by platelets from spleen to the blood and that deterioration of TPO production may play a role in thrombocytopenia in liver cirrhosis.

Elevated IgG4 levels in a case with multicentric Castleman's disease
Author:F Ishida; K Kitano; H Kobayashi; H Saito; K Kiyosawa

Eosinophilia associated with clonal T-cell proliferation
LEUKEMIA & LYMPHOMA,27(3-4):335-342 1997(Oct.)
Author:K Kitano; N Ichikawa; S Shimodaira; T Ito; F Ishida; K Kiyosawa
Abstract:Eosinophilia associated with the expansion of cloned T-cells is reviewed in relation to cytokine production. It has been proved that eosinophilopoiesis is caused by eosinophil-stimulating cytokines, including interleukin-5 (IL-5), granulocyte-macrophage colony-stimulating factor and interleukin-3, which are secreted from T-cells. Recently, we and other groups have reported several cases of eosinophilia including hypereosinophilic syndrome (HES) accompanied with proliferation of abnormal T-cells with an unusual phenotype CD3(-)CD4(+) or CD3(+)CD4(-)CD8(-) in the peripheral blood. The T-cells clonally proliferate, as confirmed by clonal rearrangements of the T-cell receptor (TCR) gene, and produce eosinophil-stimulating cytokines, especially IL-5, with or without stimulation in vitro. Although HES is defined by the combination of unexplained prolonged eosinophilia and evidence of organ involvement, these observations suggest that increased production of eosinophil-stimulating cytokines from the abnormal T-cells with phenotype CD3(-)CD4(+) or CD3(+)CD4(-)CD8(-) may cause eosinophilia, some of which have been diagnosed as HES.

Resistance to activated protein C and Arg 506 Gln factor V mutation are uncommon in eastern Asian populations
ACTA HAEMATOLOGICA,98(1):22-25 1997
Author:H Kodaira; F Ishida; S Shimodaira; O Takamiya; K Furihata; K Kitano
Abstract:We investigated the prevalence of the factor V (FV) Arg 506 Gin mutation in healthy subjects from three eastern Asian countries (Japan, n = 270; China, n = 113; and Korea, n = 93) and in 26 Japanese patients showing venous thromboembolic events. The patients were also examined for activated protein C (APC) resistance by using the Coatest(R) APC resistance kit, The FV mutation was investigated by polymerase chain reaction and restricted enzyme digestion with Mn/I RFLP assay of the FV gene, None of the patients showed APC resistance, while all subjects examined were homozygous for Arg at position 506 of the RV gene, Our results imply that FV mutation and APC resistance contribute little to venous thrombotic diseases in eastern Asia.

Serum thrombopoietin (c-Mpl ligand) levels in patients with liver cirrhosis
THROMBOSIS AND HAEMOSTASIS,76(4):545-548 1996(Oct.)
Author:S Shimodaira; F Ishida; N Ichikawa; T Tahara; T Kato; H Kodaira; T Ito; E Tanaka; T Sodeyama; K Kiyosawa; K Kitano
Abstract:To clarify the role of c-Mpl ligand (thrombopoietin: TPO) in liver cirrhosis (LC), we examined serum TPO levels (sTPO) in patients with LC (N = 44), chronic hepatitis (CH; N = 13) and healthy controls (N = 41) by an enzyme-linked immunosorbent assay. Although platelet counts of all LC patients (89 +/- 59 X 10(9)/l; mean +/- SD) were lower than those of controls and CH patients, sTPO levels in LC patients (1.23 +/- 0.51 fmol/ml) were the same as those in controls (1.22 +/- 0.37) and CH patients (1.18 +/- 0.36). Platelet counts were significantly higher in splenectomized patients than in unsplenectomized patients, but the sTPO level did not differ between these two groups. In LC patients, the sTPO level was not correlated with the platelet count, but was correlated with prothrombin time, activated partial thromboplastin time, and total bilirubin, indicating that production of TPO in the liver decreases slightly with the development of liver dysfunction. Our findings suggest that production of TPO is maintained in LC patients and their thrombocytopenia is not due to a defect in platelet production.

Serum thrombopoietin (c-Mpl ligand) levels in patients with liver cirrhosis
THROMBOSIS AND HAEMOSTASIS,76(4):545-548 1996(Oct.)
Author:S Shimodaira; F Ishida; N Ichikawa; T Tahara; T Kato; H Kodaira; T Ito; E Tanaka; T Sodeyama; K Kiyosawa; K Kitano
Abstract:To clarify the role of c-Mpl ligand (thrombopoietin: TPO) in liver cirrhosis (LC), we examined serum TPO levels (sTPO) in patients with LC (N = 44), chronic hepatitis (CH; N = 13) and healthy controls (N = 41) by an enzyme-linked immunosorbent assay. Although platelet counts of all LC patients (89 +/- 59 X 10(9)/l; mean +/- SD) were lower than those of controls and CH patients, sTPO levels in LC patients (1.23 +/- 0.51 fmol/ml) were the same as those in controls (1.22 +/- 0.37) and CH patients (1.18 +/- 0.36). Platelet counts were significantly higher in splenectomized patients than in unsplenectomized patients, but the sTPO level did not differ between these two groups. In LC patients, the sTPO level was not correlated with the platelet count, but was correlated with prothrombin time, activated partial thromboplastin time, and total bilirubin, indicating that production of TPO in the liver decreases slightly with the development of liver dysfunction. Our findings suggest that production of TPO is maintained in LC patients and their thrombocytopenia is not due to a defect in platelet production.

The largest isoform of platelet membrane glycoprotein Ib alpha is commonly distributed in Eastern Asian populations
THROMBOSIS AND HAEMOSTASIS,76(2):245-247 1996(Aug.)
Author:F Ishida; K Furihata; K Ishida; H Kodaira; KS Han; L DaZhuang; K Kitano; K Kiyosawa
Abstract:Platelet membrane glycoprotein Ib alpha has at least two polymorphisms which affect phenotype. One is the dimorphism at codon 145, and the other is a molecular weight polymorphism due to variable numbers of tandem repeats (TR) in the macroglycopeptide region. These two polymorphisms are in linkage disequilibrium. The frequencies of these polymorphisms differ considerably depending on race, and the largest variant with four TR is almost exclusively present in the Japanese population. We examined the genotypes of HPA-2 and TR polymorphism in three different races from Eastern Asia; the Japanese (n = 103), Korean (n = 101) and Chinese population (n = 177). The gene frequency of HPA-2 differed significantly among these three populations. Among HPA-2b-positive individuals, the A isoform with four TR and B with three TR were present in all three populations and A dominated over B. Individuals homozygous for the A isoform were found in both Japanese and Korean populations. These findings indicate that the largest haplotype is common in the Eastern Asian region.

Regulation of serum thrombopoietin levels by platelets and megakaryocytes in patients with aplastic anemia and idiopathic thrombocytopenic purpura.
EXPERIMENTAL HEMATOLOGY,24(9):428-428 1996(Aug.)
Author:N Ichikawa; F Ishida; S Shimodaira; T Ito; T Tahara; T Kato; K Kitano

Eosinophilia associated with proliferation of CD3(+)4(-)8(-) alpha beta(+) T cells with chromosome 16 anomalies
Author:K Kitano; N Ichikawa; B Mahbub; M Ueno; T Ito; S Shimodaira; H Kodaira; F Ishida; H Kobayashi; H Saito; Y Okubo; H Enokihara; K Kiyosawa
Abstract:We describe a patient with eosinophilia and an abnormal CD3(+)4(-)8(-)alpha beta(+) T-cell population. Chromosomal analysis of sorted CD3(+)4(-)8(-) cells revealed abnormal karyotypes on chromosome 16. In the presence of IL-2 the production of IL-5 from CD3(+)4(-)8(-) cells was higher than that from CD3(+)4(+)/8(+) cells. Eosinophil survival-enhancing activity in the patient serum was inhibited by a combination of anti-IL-5 and anti-GM-CSF monoclonal antibodies. These data suggest that increased production of IL-5 and GM-CSF from the abnormal CD3(+)4(-)8(-) cells might cause eosinophilia.

Regulation of serum thrombopoietin levels by platelets and megakaryocytes in patients with aplastic anaemia and idiopathic thrombocytopenic purpura
Thromb Haemost,76(2):156-160 1996
Author:Ichikawa N; Ishida F

THROMBOSIS AND HAEMOSTASIS,74(3):996-996 1995(Sep.)

BLOOD,86(4):1356-1360 1995(Aug.)
Abstract:Platelet membrane glycoprotein Ib alpha (GPIb alpha) bears the human platelet alloantigen (HPA)-2 and molecular weight (MW) polymorphisms on sodium dodecyl sulfate-polyacrylamide gels. HPA-2 arises from a threonine/methionine dimorphism at residue 145 of the GPIb alpha sequence, whereas different numbers of tandem repeats of a 39-bp sequence encoding 13-amino acids corresponding to a region between serine(399) and threonine(411) of the GPIb alpha account for the latter. To identify the genetic basis of the MW polymorphism among Japanese, we counted the tandem repeats in 103 individuals. In addition to the reported three variants with one, two, or three tandem repeats, we identified a new variant with four perfect tandem repeats of the 39-bp sequence that corresponded to the largest phenotype. Phenotypic analysis of the MW polymorphism on 12 individuals including all four phenotypes completely accorded in the genotype. We also determined the genotype of HPA-2 and found that methionine(145) was in complete linkage disequilibrium, with the larger variants containing three or four tandem repeats. These results imply a model of evolutionary steps in the gene encoding GPIb alpha. (C) 1995 by The American Society of Hematology.

Abstract:The clonal proliferation of large granular lymphocytes can be detected in patients with T-cell-lineage granular lymphocyte-proliferative disorders (T-GLPD) by Southern blotting T-cell receptor genes. However, this cannot be applied to patients with natural killer-cell-lineage GLPD (NK-GLPD) as it lacks a clonal marker. We therefore investigated the use of two other diagnostic techniques in evaluating clonal proliferation in Japanese patients with NK-GLPD (n=4) and T-GLPD (n=3) by chromosomal analysis of peripheral blood mononuclear cells (PBMC) stimulated with either interleukin-2 or phytohaemagglutinin, and Epstein-Barr viral (EBV) genomic DNA analysis, Chromosomal analysis revealed abnormal karyotypes in the PBMC of three of four patients with NK-GLPD, whereas EBV analysis showed a monoclonal terminal configuration in the PBMC in the fourth patient. Southern blots revealed rearrangements of the TCR genes in all three patients with T-GLPD but in none of those with NK-GLPD. It is suggested that these methods may be useful in detecting the abnormal proliferation of large granular lymphocytes in NK-GLPD.

Arg 506 Gln factor V mutation is uncommon in eastern Asian populations.
BLOOD,86(10):3658-3658 1995
Author:Kodaira, H; Ishida, F; Ito, T; Ichikawa, N; Shimofaira, S; Takamiya, O; Furihata, K; Kiyosawa, K; Kitano, K

Serum thrombopoietin (c-Mpl ligand) levels in patients with liver cirrhosis
BLOOD,86(10):3621-3621 1995
Author:Shimodaira, S; Ishida, F; Ichikawa, N; Tahara, T; Kato, T; Kodaira, H; Ito, T; Tanaka, E; Sodeyama, T; Kiyosawa, K; Kitano, K

Abstract:Cytogenetic analysis of bone marrow (BM) cells from a patient with myelodysplastic syndrome (MDS) associated with eosinophilia showed a 45,XY,t(12;21)(q23;q22), -17 karyotype. We performed clonal and suspension cultures using the patient's BM mononuclear cells to clarify the mechanism of eosinophilia. Eosinophil colonies formed in the presence of interleukin-5 (IL-5) and granulocyte-macrophage colony-stimulating factor (GM-CSF), but not in the presence of IL-3. When BM cells were cultured in suspension in the presence of IL-5, they differentiated to mature eosinophils, and chromosome analysis identified the 45,XY,t(12;21)(q23;q22), -17 karyotype in all metaphases. The patient's serum did not stimulate eosinophil proliferation or differentiation in comparison with normal serum, however, these data suggest that the abnormal clone with 45,XY,t(12;21)(q23;q22), -17 karyotype may have an increased responsiveness to IL-5 and GM-CSF, resulting in eosinophilia.

Abstract:We report a case of transfusion-associated graft-vs.-host disease (TA-GVHD) in a previously healthy, 58 year old Japanese man following an accidental injury. Despite intensive treatment, the patient died 22 days after the transfusion. Although many cases of fatal TA-GVHD have been reported in immunocompetent patients, only two are known to have developed this disorder unrelated to a surgical procedure. This is the first such case to be confirmed by HLA typing. Our report suggests that TA-GVHD can occur even in a healthy, immunocompetent recipient. Thus, it seems necessary to establish guidelines for the irradiation of cellular blood products before transfusion.

ACTA HAEMATOLOGICA,90(1):42-45 1993
Abstract:A case of 64-year-old Japanese woman who developed aplastic anemia and idiopathic thrombocytopenic purpura with antibody to platelet glycoprotein IIb/IIIa 4 years following a resection of malignant thymoma is reported. Bone marrow was hypocellular and ferrokinetics revealed the prolongation of the half-time of plasma iron disappearance and a decrease in red cell utilization, findings compatible with a diagnosis of aplastic anemia. The life span of platelets was markedly decreased to 3.07 h, and the test for antiplatelet glycoprotein IIb/IIIa antibody was positive. The patient's serum had a suppressive effect on the formation of colonies of burst-forming unit-erythroid and colony-forming unit-granulocyte using normal bone marrow cells. These results suggest that the aplastic anemia in this patient may have been induced by some suppressive activity in the serum. Splenectomy followed by an administration of cyclosporine effectively restored the peripheral blood count.

BLOOD,79(11):3086-3090 1992(Jun.)

BLOOD,78(7):1722-1729 1991(Oct.)

THROMBOSIS AND HAEMOSTASIS,65(6):796-796 1991(Jun.)

IL-2添加による染色体分析で47XY, +8の異常を認めたnatural killer細胞型granular lymphocyte leukemia
臨床血液,31(10):1711-1715 1990
Author:石田 文宏; 斉藤 博; 降旗 謙一; 古田 精市; 園山 政行; 押味 和夫

Mutational Landscape of Aggressive Natural Killer Cell Leukemia and Drug Sensitivity Profiling Reveal Therapeutic Options in Natural Killer Cell Malignancies
BLOOD,128(22) 2016(Dec.)
Author:Olli Dufva; Matti Kankainen; Tiina Kelkka; Shady Adnan Awad; Nodoka Sekiguchi; Bhagwan Yadav; Samuli Eldfors; Heikki Kuusanmaki; Emma I. Andersson; Disha Malani; Paavo Pietarinen; Leena Saikko; Panu E. Kovanen; Junji Suzumiya; Ritsuro Suzuki; Young Hyeh Ko; Won Seog Kim; Shih-Sung Chuang; Thomas P. Loughran; Wing Chung Chan; Koichi Ohshima; Tero Aittokallio; Fumihiro Ishida; Satu Mustjoki

Exome Sequencing of Aggressive Natural Killer Cell Leukemia and Drug Profiling Highlight Candidate Driver Pathways in Malignant Natural Killer Cells
BLOOD,126(23) 2015(Dec.)
Author:Olli Dufva Bmed; Tiina Kelkka; Shady Awad; Nodoka Sekiguchi; Heikki Kuusanmaki; Emma I. Andersson; Samuli Eldfors; Disha Malani; Paavo Pietarinen; Leena Saikko; Panu E. Kovanen; Junji Suzumiya; Koichi Ohshima; Young Hyeh Ko; Won Seog Kim; Shih-Sung Chuang; Thomas P. Loughran; Wing C. Chan; Fumihiro Ishida; Satu Mustjoki

STAT5b Gene Mutations and Their Association with a Unique Immunophenotype of CD4+CD56+TCR alpha beta Type LGL Leukemia
BLOOD,124(21) 2014(Dec.)
Author:Takahiro Tanahashi; Kazuyuki Matsuda; Nodoka Sekiguchi; Sayaka Nishina; Noriko Senoo; Kaoko Sakai; Hitoshi Sakai; Toshiro Ito; Takeki Mitsui; Yok Lam Kwong; Fumihiro Ishida

Clinicopathological features and treatments of aggressive NK-cell leukemia
血液内科,68(1):124-128 2014(Jan.)
Author:石田 文宏

Novel Prognostic Model Of Primary Mediastinal Large B-Cell Lymphoma (PMBL): A Multicenter Cooperative Retrospective Study In Japan
BLOOD,122(21) 2013(Nov.)
Author:Tomohiro Aoki; Koji Izutsu; Ritsuro Suzuki; Chiaki Nakaseko; Hiroshi Arima; Kazuyuki Shimada; Makoto Sasaki; Jun Takizawa; Kinuko Mitani; Tadahiko Igarashi; Yoshinobu Maeda; Fumihiro Ishida; Nozomi Niitsu; Ken Ohmachi; Hirotaka Takasaki; Naoya Nakamura; Tomohiro Kinoshita; Shigeo Nakamura; Michinori Ogura

STAT3 Gene Mutations and the Association with Pure Red Cell Aplasia in Large Granular Lymphocyte Leukemia.
BLOOD,120(21) 2012(Nov.)
Author:Nodoka Sekiguchi; Kazuyuki Matsuda; Kayoko Momose; Hideki Makishima; Toshiro Ito; Fumihiro Ishida

Aggressive NK Cell Leukemia (ANKL): Experience with 34 Patients and Therapeutic potentials of L-Asparaginase and Allogeneic Hematopoietic Cell Transplantation - A Japan-Korea Multicenter Study for ANKL (ANKL07)
BLOOD,116(21):1274-1275 2010(Nov.)
Author:Fumihiro Ishida; Won Seog Kim; Young Hyeh Ko; Junji Suzumiya; Yasushi Isobe; Kazuo Oshimi; Shigeo Nakamura; Ritsuro Suzuki

Pretreatment EBV-DNA Copy Number Is Predictive for Response to SMILE Chemotherapy for Newly-Diagnosed Stage IV, Relapsed or Refractory Extranodal NK/T-Cell Lymphoma, Nasal Type: Results of NKTSG Phase II Study
BLOOD,116(21):1183-1184 2010(Nov.)
Author:Ritsuro Suzuki; Hiroshi Kimura; Yok-Lam Kwong; Yoshinobu Maeda; Chizuko Hashimoto; Won Seog Kim; Cheolwon Suh; Koji Izutsu; Fumihiro Ishida; Yoshinori Ito; Motoko Yamaguchi; Junji Suzumiya; Rie Hyo; Shigeo Nakamura; Kazuo Oshimi

Profiles of Cytotoxic Molecules In Mature NK Cell Neoplasms
BLOOD,116(21):1685-1685 2010(Nov.)
Author:Nodoka Sekeguchi; Naoko Asano; Hideki Makishima; Masanobu Momose; Toshiro Ito; Shigeo Nakamura; Fumihiro Ishida

Author:F. Ishida; Y. H. Ko; W. S. Kim; J. Suzumiya; Y. Isobe; K. Oshimi; S. Nakamura; R. Suzuki

Linkage of expression of chemokine receptors (CXCR3 and CCR4) and cytotoxic molecules in peripheral T cell lymphoma, not otherwise specified and ALK-negative anaplastic large cell lymphoma.
Int J Hematol,91(3):426-435 2010
Author:Asano N; Suzuki R; Ohshima K; Kagami Y; Ishida F; Yoshino T; Ogawa H; Morishima Y; Nakamura S

Cytotoxic Molecule (CM)-Positive Classical Hodgkin Lymphoma: a Clinicopathologic Study in Comparison with Nodal Peripheral T-Cell Lymphoma of Not Otherwise Specified Type Possessing CM Expression.
BLOOD,114(22):620-620 2009(Nov.)
Author:Naoko Asano; Jun-Ichi Tamaru; Fumihiro Ishida; Tadashi Yoshino; Yoshitoyo Kagami; Yasuo Morishima; Masao Seto; Tomohiro Kinoshita; Shigeo Nakamura

診断に苦慮したCD4+/CD56+ hematodermic neoplasm(blastic NK cell lymphoma)の1例
Skin Cancer,23(2):238-238 2008(Nov.)
Author:木藤 健治; 鶴田 恭子; 生垣 英之; 塩原 順子; 木庭 幸子; 高沢 裕子; 村田 浩; 太田 桂子; 高田 実; 斎田 俊明; 石田 文宏; 伊藤 俊郎; 中澤 英之; 福島 万奈; 浅野 直子; 中村 栄男
Abstract:A 63-year-old man had noticed a nodule on the lower back about 4 months before consultation. He visited a clinic nearby his house and malignant lymphoma was suspected. He was subsequently referred to our department. Histopathologic analysis revealed proliferation of small to medium-sized atypical lymphocytes. The tumor cells had a CD3 (+), CD4 (+), CD20 (+) phenotype, thereby diagnosed as &ldquo;peripheral T-cell lymphoma, unspecified&rdquo;. He received 6 courses of CHOP regimen, leading to a complete remission (CR) . However, about 11 months after the therapy, recurrence of lymphoma on the trunk and extremities was confirmed histopathologically by skin biopsy of the erythematous lesions. The biopsy specimen showed infiltration of monotonous atypical lymphocytes with a phenotype of CD4 (+), CD79a (-), CD56 (+), CD123 (+), TdT (-), MPO (-), and EBV-ISH (-), establishing the diagnosis of &ldquo;CD4+/CD56+ hematodermic neoplasm (blastic NK cell lymphoma)&rdquo;. He received 2 courses of CHOP regimen and then 1 course of hyper-CVAD regimen; thereafter he underwent allogenic peripheral blood stem cell transplantation, leading to a CR. He has remained in CR without any symptoms for about 6 months. [<i>Skin Cancer (Japan)</i>2008; 23: 410-415]

診断に苦慮したCD4+/CD56+ hematodermic neoplasm(blastic NK cell lymphoma)の一例
日本皮膚悪性腫瘍学会学術大会プログラム・抄録集,24回:182-182 2008(Jul.)
Author:木藤 健治; 鶴田 恭子; 生垣 英之; 塩原 順子; 木庭 幸子; 高沢 裕子; 村田 浩; 太田 桂子; 高田 実; 斎田 俊明; 石田 文宏; 伊藤 俊郎; 中澤 英之; 福島 万奈; 浅野 直子; 中村 栄男

Expression of chemokine receptors (CXCR3 and CCR4) and cytotoxic molecules in peripheral T-cell lymphoma, unspecified and ALK-negative anaplastic large cell lymphoma
ANNALS OF ONCOLOGY,19:154-154 2008(Jun.)
Author:N. Asano; R. Suzuki; K. Ohshima; F. Ishida; Y. Kagami; T. Yoshino; Y. Morishima; S. Nakamura

Chemokine system in NK-cell neoplasms
血液・腫瘍科,54(3):367-372 2007(Mar.)
Author:牧島 秀樹; 石田 文宏

Expression levels of CXCR1 and CCR5 on natural killer (NK) cells in aggressive NK cell leukemia and chronic NK cell lymphocytosis.
BLOOD,102(11):604A-604A 2003(Nov.)
Author:H Makishima; F Ishida; T Ito; S Shimodaira; K Kiyosawa

Identification of novel molecular markers in T cell type lymphoproliferative disease of granular lymphocytes by microarray analysis with purified T cell populations.
BLOOD,98(11):162B-162B 2001(Nov.)
Author:H Makishima; F Ishida; K Kitano; K Kiyosawa; K Omine; H Mano

Skewed T cell receptor repertoires with oligoclonal profiles in patients with CD8-positive T cell lymphoproliferative disease of granular lymphocytes.
BLOOD,96(11):127A-127A 2000(Nov.)
Author:H Makishima; F Ishida; T Ito; M Ota; Y Katsuyama; M Ueno; S Shimodaira; K Kitano; K Kiyosawa