Shinshu University HOMEJAPANESEAccess / Campus Map

Shinshu University Researcher DirectoryShinshu University Researcher Directory

Search by Researcher / Research Field
Search by Category

Sato Takashi

Academic OrganizationFaculty of AgricultureTEL81-265-77-1403
Education and Research OrganizationInterdisciplinary Cluster for Cutting Edge Research, Institute for Biomedical SciencesFAX81-265-77-1403
PositionSpecially Appointed ProfessorMail Address
Address8304, Minamiminowa-Village, Kamiina-County 399-4598Web site

Modified:03/06/2024

Profile

Research Field
Respiratory medicine
Academic Societies
Academic Societies
日本臨床腫瘍学会
日本感染症学会
日本化学療法学会
日本アレルギー学会
日本酪農科学会
日本呼吸器学会
日本内科学会
American Thoracic Society

Committee of Academic Societies
2012-2015 , American Thoracic Society , Program Committee Member
2008-2010 , American Thoracic Society , Program Committee Member
Academic Background
Graduate School
Yokohama City University , (病態免疫制御内科学 , 医学研究科) , 2006

College
Yokohama City University , (医学部) , 2000
Tokyo Institute of Technology , (工学部) , 1994

Degree
Ph.D , Yokohama City University (Japan)
Awards
2013 , NIH Fellows Award for Research Excellence (FARE) 2014
2013 , 米国国立衛生研究所(NIH)Appreciation Award
2009 , 横浜市立大学第一内科同門会賞
2008 , 神奈川難病財団研究奨励賞
2007 , The Oak Ridge Institute for Science and Education (ORISE) Research Award
2007 , 横浜総合医学振興財団呼吸器疾患研究賞
Research Career
Research Career
2022- , 富士電機株式会社 半導体事業本部, 松本工場, 専属産業医
2020- , Department of Pulmonology, School of Medicine, Yokohama City University
2019- , Interdisciplinary Cluster for Cutting Edge Research, Shinshu University
2020-2022 , Head of Medical Oncology Department/Director of Research Center, Tokyo Shinagawa Hospital
2019-2019 , Faculty of Agriculture, Shinshu University

Overseas Education
2012-2014 , 米国国立衛生研究所(NIH)国立がん研究所(NCI)
2007-2008 , 米国国立衛生研究所(NIH)国立がん研究所(NCI)
2005-2006 , 米国食品医薬品局(FDA)生物製剤評価研究センター(CBER)

Research

Books, Articles, etc.
Books
クリニカルトピックス, Innovative therapy for lung cancer - intratracheal delivery of probiotics harboring immune checkpoint-blocking single-chain variable fragment -
BIO Clinica 2023;38(7):638-42 2023(Jun. 28)
Author:Takashi Sato


Chapter title: Intratracheally therapeutic option for COPD: a potential usage of the therapeutic microbe for delivering specific protein to the lungs, Book title: Chronic Obstructive Pulmonary Disease - A Compendium of Medicine and the Humanities
IntechOpen 2022(Aug. 17)
Author:Takashi Sato; Takeshi Shimosato
Abstract:Abstract Currently, inhaled therapy using corticosteroids and/or bronchodilators is the major established treatment for chronic obstructive pulmonary disease (COPD). The topic to be covered in this chapter is the recently developed experimental approach using biologically active molecules secreted by the live genetically modified lactic acid bacteria (gmLAB). The strategy to use gmLAB as a therapeutic/delivering tool targeting disease-specific active molecules/cites is proceeding. The role of inflammation and oxidative stress in COPD development is a valid target point. Heme oxygenase (HO)-1 as an anti-inflammatory and antioxidative stress molecule has been examined to attenuate the lung function decline and inflammation in the murine model of COPD. Recently, HO-1-secreting gmLAB as a tool for targeting inflammatory diseases has been developed and examined in several disease models including COPD. When administered intratracheally, the gmLAB showed migration to the peripheral lung and overexpression of anti-inflammatory/oxidative HO-1 in both lung and serum, protecting the lung from COPD development.


Development of a New Treatment Modality for Lung Diseases That Uses Innovative Fine Droplet Drying (FDD) Technology Offering Inhalable Nano/microparticles-incorporated Therapeutic Agents
Science Impact Ltd 2019(Mar.)
Author:SATO Takashi; SHIMOSATO Takeshi
Abstract:DOI:https://doi.org/10.21820/23987073.2019.3.56


Chapter 14: Pharmacokinetics, Pharmacodynamics, and Toxicities: What Should We Know About Genetic Factors that Affect the Pharmacotherapy of Pulmonary Diseases?, Respiratory Disease Series: Diagnostic Tools and Disease Managements, 255-271
Springer 2018(Jul.)
Author:SATO Takashi


第40章 肺胞を拡散する酸素と二酸化炭素のガス交換の原理、第41章 血液や組織液中の酸素と二酸化炭素の輸送, ガイトン生理学(原著第13版)
ELSEVIER 2018(Mar.)
Author:佐藤 隆


Articles
Oral administration of Brevibacterium linens from washed cheese increases the proportions of short‐chain fatty acid‐producing bacteria and lactobacilli in the gut microbiota of mice
Animal Science Journal,94(1) 2023(Dec. 16)
Author:Aito Murakami; Koharu Toyomoto; Fu Namai; Takashi Sato; Tadashi Fujii; Takumi Tochio; Takeshi Shimosato
Abstract:Abstract Brevibacterium linens (B. linens) is a dairy microorganism used in the production of washed cheese. However, there has been little research on B. linens, especially regarding its effects in vivo. Herein, we report the morphological characteristics of B. linens, such as its two‐phase growth and V‐ and Y‐shaped bodies. We also report that oral administration of B. linens increased the diversity of the gut microbiota and promoted the growth of lactobacilli and short‐chain fatty acid‐producing bacteria, such as Lachnospiraceae and Muribaculaceae. These findings suggest that the ingestion of B. linens may have beneficial effects in humans and animals.


CD62L expression marks SARS-CoV-2 memory B cell subset with preference for neutralizing epitopes
Science Advances,9(24) 2023(Jun. 16)
Author:Taishi Onodera; Nicolas Sax; Takashi Sato; Yu Adachi; Ryutaro Kotaki; Takeshi Inoue; Ryo Shinnakasu; Takayuki Nakagawa; Shuetsu Fukushi; Tommy Terooatea; Mai Yoshikawa; Keisuke Tonouchi; Takaki Nagakura; Saya Moriyama; Takayuki Matsumura; Masanori Isogawa; Kazutaka Terahara; Tomohiro Takano; Lin Sun; Ayae Nishiyama; Shinnya Omoto; Masaharu Shinkai; Tomohiro Kurosaki; Kazuo Yamashita; Yoshimasa Takahashi
Abstract:Severe acute respiratory syndrome coronavirus 2–neutralizing antibodies primarily target the spike receptor binding domain (RBD). However, B cell antigen receptors (BCRs) on RBD-binding memory B (B mem ) cells have variation in the neutralizing activities. Here, by combining single B mem cell profiling with antibody functional assessment, we dissected the phenotype of B mem cell harboring the potently neutralizing antibodies in coronavirus disease 2019 (COVID-19)–convalescent individuals. The neutralizing subset was marked by an elevated CD62L expression and characterized by distinct epitope preference and usage of convergent V H (variable region of immunoglobulin heavy chain) genes, accounting for the neutralizing activities. Concordantly, the correlation was observed between neutralizing antibody titers in blood and CD62L + subset, despite the equivalent RBD binding of CD62L + and CD62L subset. Furthermore, the kinetics of CD62L + subset differed between the patients who recovered from different COVID-19 severities. Our B mem cell profiling reveals the unique phenotype of B mem cell subset that harbors potently neutralizing BCRs, advancing our understanding of humoral protection.


High Fever, Wide Distribution of Viral Pneumonia, and Pleural Effusion are More Critical Findings at the First Visit in Predicting the Prognosis of COVID-19: A Single Center, retrospective, Propensity Score-Matched Case–Control Study
International Journal of General Medicine,Volume 16:2337-2348 2023(Jun. 08)
Author:Masahiro Shinoda; Shinichiro Ota; Yuto Yoshida; Takatomo Hirouchi; Kanako Shinada; Takashi Sato; Miwa Morikawa; Naoki Ishii; Masaharu Shinkai


Development of a Single-Chain Fragment Variable that Binds to the SARS-CoV-2 Spike Protein Produced by Genetically Modified Lactic Acid Bacteria
Molecular Biotechnology 2023(Apr. 15)
Author:Suzuka Oshima; Fu Namai; Takashi Sato; Takeshi Shimosato


Heterologous SARS-CoV-2 spike protein booster elicits durable and broad antibody responses against the receptor-binding domain.
Nature communications,14(1):1451-1451 2023(Mar. 15)
Author:Tomohiro Takano; Takashi Sato; Ryutaro Kotaki; Saya Moriyama; Shuetsu Fukushi; Masahiro Shinoda; Kiyomi Kabasawa; Nagashige Shimada; Mio Kousaka; Yu Adachi; Taishi Onodera; Kazutaka Terahara; Masanori Isogawa; Takayuki Matsumura; Masaharu Shinkai; Yoshimasa Takahashi
Abstract:The immunogenicity of mRNA vaccines has not been well studied when compared to different vaccine modalities in the context of additional boosters. Here we show that longitudinal analysis reveals more sustained SARS-CoV-2 spike receptor-binding domain (RBD)-binding IgG titers with the breadth to antigenically distinct variants by the S-268019-b spike protein booster compared to the BNT162b2 mRNA homologous booster. The durability and breadth of RBD-angiotensin-converting enzyme 2 (ACE2) binding inhibitory antibodies are pronounced in the group without systemic adverse events (AEs) after the S-268019-b booster, leading to the elevated neutralizing activities against Omicron BA.1 and BA.5 variants in the stratified group. In contrast, BNT162b2 homologous booster elicited antibodies to spike N-terminal domain in proportion to the AE scores. High-dimensional immune profiling identifies early CD16+ natural killer cell dynamics with CCR3 upregulation, as one of the correlates for the distinct anti-RBD antibody responses by the S-268019-b booster. Our results illustrate the combinational effects of heterologous booster on the immune dynamics and the durability and breadth of recalled anti-RBD antibody responses against emerging virus variants.


Development of fluorescence-labeled antibody for immune checkpoint inhibitor using engineered probiotics
AMB Express,13(1) 2023(Jan. 12)
Author:Fu Namai; Shunsuke Sumiya; Natsumi Nomura; Takashi Sato; Takeshi Shimosato
Abstract:Abstract Here, we developed a genetically modified lactic acid bacteria (gmLAB) that produces green fluorescent protein (GFP)-conjugating, anti-programmed death-ligand 1 (PD-L1) single-chain variable fragments (scFv) for use as an anti-cancer device that targets immune checkpoint molecules. Since PD-L1 plays a key role as an immune checkpoint molecule in the tumor microenvironment, inhibition and detection of PD-L1 are important in cancer research. The anti-PD-L1 scFv was designed based on atezolizumab, a humanized IgG1 monoclonal antibody, and integrated into a lactococcal GFP gene expression vector. Gene expression from the constructed gmLAB was confirmed by western blotting and GFP fluorescence. The ability of GFP-conjugating anti-PD-L1 scFv against the target antigen, PD-L1 protein, was shown using an enzyme-linked immunosorbent assay. Finally, the ability to recognize PD-L1-expressing tumor-cell lines was confirmed using flow cytometry and fluorescence microscopy. Our results suggest that the gmLAB could be applied to in vivo imaging in cancer as an affordable diagnostic/treatment tool. Graphical Abstract


SARS-CoV-2 specific CD4+ T cell longevity correlates with Th17-like phenotype
iScience,:104959-104959 2022(Aug. 17)
Author:Kazutaka Terahara; Takashi Sato; Yu Adachi; Keisuke Tonouchi; Taishi Onodera; Saya Moriyama; Lin Sun; Tomohiro Takano; Ayae Nishiyama; Ai Kawana-Tachikawa; Tetsuro Matano; Takayuki Matsumura; Masaharu Shinkai; Masanori Isogawa; Yoshimasa Takahashi


Immunogenicity and safety of booster dose of S-268019-b or BNT162b2 in Japanese participants: An interim report of phase 2/3, randomized, observer-blinded, noninferiority study
Vaccine 2022(Jun. 20)
Author:Masaharu Shinkai; Takuhiro Sonoyama; Akari Kamitani; Risa Yokokawa Shibata; Naomi M. Seki; Shinya Omoto; Masahiro Shinoda; Takashi Sato; Naoki Ishii; Kenji Igarashi; Mari Ariyasu


Longitudinal Analysis of Neutralizing Potency against SARS-CoV-2 in the Recovered Patients after Treatment with or without Favipiravir
Viruses,14(4):670-670 2022(Mar. 24)
Author:Kanako Shinada; Takashi Sato; Saya Moriyama; Yu Adachi; Masahiro Shinoda; Shinichiro Ota; Miwa Morikawa; Masamichi Mineshita; Takayuki Matsumura; Yoshimasa Takahashi; Masaharu Shinkai
Abstract:The effect of treatment with favipiravir, an antiviral purine nucleoside analog, for coronavirus disease 2019 (COVID-19) on the production and duration of neutralizing antibodies for SARS-CoV-2 was explored. There were 17 age-, gender-, and body mass index-matched pairs of favipiravir treated versus control selected from a total of 99 patients recovered from moderate COVID-19. These subjects participated in the longitudinal (>6 months) analysis of (i) SARS-CoV-2 spike protein’s receptor-binding domain IgG, (ii) virus neutralization assay using authentic virus, and (iii) neutralization potency against original (WT) SARS-CoV-2 and cross-neutralization against B.1.351 (beta) variant carrying triple mutations of K417N, E484K, and N501Y. The results demonstrate that the use of favipiravir: (1) significantly accelerated the elimination of SARS-CoV-2 in the case vs. control groups (p = 0.027), (2) preserved the generation and persistence of neutralizing antibodies in the host, and (3) did not interfere the maturation of neutralizing potency of anti-SARS-CoV-2 and neutralizing breadth against SARS-CoV-2 variants. In conclusion, treatment of COVID-19 with favipiravir accelerates viral clearance and does not interfere the generation or maturation of neutralizing potency against both WT SARS-CoV-2 and its variants.


Glycan engineering of the SARS-CoV-2 receptor-binding domain elicits cross-neutralizing antibodies for SARS-related viruses
Journal of Experimental Medicine,218(12) 2021(Dec. 06)
Author:Ryo Shinnakasu; Shuhei Sakakibara; Hiromi Yamamoto; Po-hung Wang; Saya Moriyama; Nicolas Sax; Chikako Ono; Atsushi Yamanaka; Yu Adachi; Taishi Onodera; Takashi Sato; Masaharu Shinkai; Ryosuke Suzuki; Yoshiharu Matsuura; Noritaka Hashii; Yoshimasa Takahashi; Takeshi Inoue; Kazuo Yamashita; Tomohiro Kurosaki
Abstract:Broadly protective vaccines against SARS-related coronaviruses that may cause future outbreaks are urgently needed. The SARS-CoV-2 spike receptor-binding domain (RBD) comprises two regions, the core-RBD and the receptor-binding motif (RBM); the former is structurally conserved between SARS-CoV-2 and SARS-CoV. Here, in order to elicit humoral responses to the more conserved core-RBD, we introduced N-linked glycans onto RBM surfaces of the SARS-CoV-2 RBD and used them as immunogens in a mouse model. We found that glycan addition elicited higher proportions of the core-RBD–specific germinal center (GC) B cells and antibody responses, thereby manifesting significant neutralizing activity for SARS-CoV, SARS-CoV-2, and the bat WIV1-CoV. These results have implications for the design of SARS-like virus vaccines.


Identification of conserved SARS-CoV-2 spike epitopes that expand public cTfh clonotypes in mild COVID-19 patients
Journal of Experimental Medicine,218(12) 2021(Dec. 06)
Author:Xiuyuan Lu; Yuki Hosono; Masamichi Nagae; Shigenari Ishizuka; Eri Ishikawa; Daisuke Motooka; Yuki Ozaki; Nicolas Sax; Yuichi Maeda; Yasuhiro Kato; Takayoshi Morita; Ryo Shinnakasu; Takeshi Inoue; Taishi Onodera; Takayuki Matsumura; Masaharu Shinkai; Takashi Sato; Shota Nakamura; Shunsuke Mori; Teru Kanda; Emi E. Nakayama; Tatsuo Shioda; Tomohiro Kurosaki; Kiyoshi Takeda; Atsushi Kumanogoh; Hisashi Arase; Hironori Nakagami; Kazuo Yamashita; Yoshimasa Takahashi; Sho Yamasaki
Abstract:Adaptive immunity is a fundamental component in controlling COVID-19. In this process, follicular helper T (Tfh) cells are a subset of CD4+ T cells that mediate the production of protective antibodies; however, the SARS-CoV-2 epitopes activating Tfh cells are not well characterized. Here, we identified and crystallized TCRs of public circulating Tfh (cTfh) clonotypes that are expanded in patients who have recovered from mild symptoms. These public clonotypes recognized the SARS-CoV-2 spike (S) epitopes conserved across emerging variants. The epitope of the most prevalent cTfh clonotype, S864–882, was presented by multiple HLAs and activated T cells in most healthy donors, suggesting that this S region is a universal T cell epitope useful for booster antigen. SARS-CoV-2–specific public cTfh clonotypes also cross-reacted with specific commensal bacteria. In this study, we identified conserved SARS-CoV-2 S epitopes that activate public cTfh clonotypes associated with mild symptoms.


Free feeding of CpG-oligodeoxynucleotide particles prophylactically attenuates allergic airway inflammation and hyperresponsiveness in mice
Frontiers in Immunology 2021(Nov. 02)
Author:Takuma Okajima, Suguru Shigemori, Fu Namai, Tasuku Ogita, Takashi Sato and Takeshi Shimosato


Temporal maturation of neutralizing antibodies in COVID-19 convalescent individuals improves potency and breadth to circulating SARS-CoV-2 variants
Immunity,54(8):1841-1852.e4 2021(Aug.)
Author:Saya Moriyama; Yu Adachi; Takashi Sato; Keisuke Tonouchi; Lin Sun; Shuetsu Fukushi; Souichi Yamada; Hitomi Kinoshita; Kiyoko Nojima; Takayuki Kanno; Minoru Tobiume; Keita Ishijima; Yudai Kuroda; Eun-Sil Park; Taishi Onodera; Takayuki Matsumura; Tomohiro Takano; Kazutaka Terahara; Masanori Isogawa; Ayae Nishiyama; Ai Kawana-Tachikawa; Masaharu Shinkai; Natsuo Tachikawa; Shigeki Nakamura; Takahiro Okai; Kazu Okuma; Tetsuro Matano; Tsuguto Fujimoto; Ken Maeda; Makoto Ohnishi; Takaji Wakita; Tadaki Suzuki; Yoshimasa Takahashi
Abstract:Antibody titers against SARS-CoV-2 slowly wane over time. Here, we examined how time affects antibody potency. To assess the impact of antibody maturation on durable neutralizing activity against original SARS-CoV-2 and emerging variants of concern (VOCs), we analyzed receptor binding domain (RBD)-specific IgG antibodies in convalescent plasma taken 1-10 months after SARS-CoV-2 infection. Longitudinal evaluation of total RBD IgG and neutralizing antibody revealed declining total antibody titers but improved neutralization potency per antibody to original SARS-CoV-2, indicative of antibody response maturation. Neutralization assays with authentic viruses revealed that early antibodies capable of neutralizing original SARS-CoV-2 had limited reactivity toward B.1.351 (501Y.V2) and P.1 (501Y.V3) variants. Antibodies from late convalescents exhibited increased neutralization potency to VOCs, suggesting persistence of cross-neutralizing antibodies in plasma. Thus, maturation of the antibody response to SARS-CoV-2 potentiates cross-neutralizing ability to circulating variants, suggesting that declining antibody titers may not be indicative of declining protection.


Oral Administration of Flavonifractor plautii, a Bacteria Increased With Green Tea Consumption, Promotes Recovery From Acute Colitis in Mice via Suppression of IL-17
Frontiers in Nutrition,7 2021(Feb. 04)
Author:Ayane Mikami; Tasuku Ogita; Fu Namai; Suguru Shigemori; Takashi Sato; Takeshi Shimosato
Abstract:Flavonifractor plautii (FP) has been reported to participate in the metabolism of catechins in the human gut. However, there is limited information on the immune regulatory effects of this bacterium. We confirmed that the administration of green tea increases the abundance of FP in the gut microbiota and investigated the effect of FP in a mouse colitis model. Mice were orally administered FP for 10 consecutive days; colonic inflammation was evaluated daily on the basis of stool consistency, gross rectal bleeding, and body weight. In the dextran sodium sulfate model, FP-exposed animals exhibited lower levels of inflammation and strong inhibition of interleukin (IL)-17 signaling. Moreover, lipoteichoic acid from FP was identified as the active component mediating IL-17 suppression. Thus, oral administration of FP appears to modulate gut inflammation and represents a viable and inexpensive oral microbial therapeutic.


Construction of Genetically Modified Lactococcus lactis Producing Anti-human-CTLA-4 Single-Chain Fragment Variable
Molecular Biotechnology,62(11-12):572-579 2020(Dec.)
Author:Fu Namai; Aito Murakami; Asami Ueda; Masami Tsukagoshi; Suguru Shigemori; Tasuku Ogita; Takashi Sato; Takeshi Shimosato


Clinical Features of 154 COVID-19 Patients and the Parameters for the Effective Detection of Pneumonia at the Time of the Initial Diagnosis in Japan
Internal Medicine 2020(Nov.)
Author:Miwa Morikawa; Masahiro Shinoda; Shinichiro Ohta; Yuto Yoshida; Takatomo Hirouchi; Kanako Shinada; Osamu Sasaki; Takashi Sato; Kenichi Kamachi; Masaharu Shinkai


Nasally Administered Lactococcus lactis Secreting Heme Oxygenase-1 Attenuates Murine Emphysema
Antioxidants,9(11):1049-1049 2020(Oct. 27)
Author:Kentaro Yumoto; Takashi Sato; Kentaro Nakashima; Fu Namai; Suguru Shigemori; Takeshi Shimosato; Takeshi Kaneko
Abstract:Emphysema, a type of lung-destroying condition associated with chronic obstructive pulmonary disease (COPD), is an inflammatory lung disease mainly due to cigarette smoke exposure. As there is no curative therapy, prevention should be considered first by cessation of smoking to avoid exposure to oxidative stresses and inflammatory mediators. In addition, therapies involving antioxidative and/or anti-inflammatory agents such as heme oxygenase (HO)-1 are candidate treatments. We developed a new tool using genetically modified Lactococcus lactis to deliver recombinant HO-1 to the lungs. Using an elastase-induced emphysema model mimicking COPD, we evaluated the effect of nasally administered L. lactis secreting HO-1 (HO-1 lactis) on cellular and molecular responses in the lungs and further disease progression. Nasally administered HO-1 lactis resulted in (1) overexpression of HO-1 in the lungs and serum and (2) attenuation of emphysema progression evaluated both physiologically and morphologically. There was a transient 5–10% weight loss compared to baseline through trafficking to the lungs when administering 1.0 × 109 cells/mouse; however, this did not impact either survival or final body weight. These results suggest that delivering HO-1 using genetically modified L. lactis through the airways could be a safe and potentially effective therapeutic approach for COPD.


Oral administration of Flavonifractor plautii attenuates inflammatory responses in obese adipose tissue
Molecular Biology Reports,47(9):6717-6725 2020(Sep.)
Author:Ayane Mikami; Tasuku Ogita; Fu Namai; Suguru Shigemori; Takashi Sato; Takeshi Shimosato


Microbial therapeutics for acute colitis based on genetically modified Lactococcus lactis hypersecreting IL-1Ra in mice
Experimental & Molecular Medicine,52(9):1627-1636 2020(Sep.)
Author:Fu Namai; Suguru Shigemori; Tasuku Ogita; Takashi Sato; Takeshi Shimosato
Abstract:Abstract The increased incidence of inflammatory bowel disease (IBD) in Western and rapidly Westernizing developing countries poses a global pandemic threat. The development of affordable drugs for treating IBD worldwide is thus a priority. Genetically modified lactic acid bacteria (gmLAB) as microbial therapeutics are inexpensive protein producers suitable for use as carriers of protein to the intestinal mucosa. Here, we successfully constructed gmLAB hypersecreting interleukin 1 receptor antagonist (IL-1Ra). Oral administration of these gmLAB suppressed body weight reduction and exacerbation of the disease activity index score in mice with acute colitis and decreased the number of CD4+ IL-17A+ cells in the mesenteric lymph nodes. These data suggest that the gmLAB deliver IL-1Ra to the colon, where it inhibits IL-1 signaling. We thus developed a novel IBD therapeutic that blocks IL-1 signaling using a gmLAB protein delivery system. This system could be an inexpensive oral microbial therapeutic.


Construction of genetically modified Lactococcus lactis that produces bioactive anti-interleukin-4 single-chain fragment variable
Molecular Biology Reports,47(9):7039-7047 2020(Sep.)
Author:Fu Namai; Suguru Shigemori; Tasuku Ogita; Takashi Sato; Takeshi Shimosato


Ribosome-Engineered Lacticaseibacillus rhamnosus Strain GG Exhibits Cell Surface Glyceraldehyde-3-Phosphate Dehydrogenase Accumulation and Enhanced Adhesion to Human Colonic Mucin
Applied and Environmental Microbiology,86(20) 2020(Aug. 14)
Author:Minori Ishida; Fu Namai; Suguru Shigemori; Shoko Kajikawa; Masami Tsukagoshi; Takashi Sato; Tasuku Ogita; Takeshi Shimosato
Abstract:ABSTRACT Differences in individual host responses have emerged as an issue regarding the health benefits of probiotics. Here, we applied ribosome engineering (RE) technology, developed in an actinomycete study, to Lacticaseibacillus rhamnosus GG (LGG). RE can effectively enhance microbial potential by using antibiotics to induce spontaneous mutations in the ribosome and/or RNA polymerase. In this study, we identified eight types of streptomycin resistance mutations in the LGG rpsL gene, which encodes ribosomal protein S12. Notably, LGG harboring the K56N mutant (LGG-MTK56N) expressed high levels of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) on the cell surface compared with the LGG wild type (LGG-WT). GAPDH plays a key role in colonic mucin adhesion. Indeed, LGG-MTK56N significantly increased type A human colonic mucin adhesion compared to LGG-WT in experiments using the Biacore system. The ability to adhere to the colon is an important property of probiotics; thus, these results suggest that RE is an effective breeding strategy for probiotic lactic acid bacteria. IMPORTANCE We sought to apply ribosome engineering (RE) to probiotic lactic acid bacteria and to verify RE’s impact. Here, we showed that one mutant of RE Lacticaseibacillus rhamnosus GG (LGG-MTK56N) bore a GAPDH on the cell surface; the GAPDH was exported via an ABC transporter. Compared to the wild-type parent, LGG-MTK56N adhered more strongly to human colonic mucin and exhibited a distinct cell size and shape. These findings demonstrate that RE in LGG-MTK56N yielded dramatic changes in protein synthesis, protein transport, and cell morphology and affected adherence to human colonic mucin.


The clinical significance of CXCL16 in the treatment of advanced non‐small cell lung cancer
Thoracic Cancer,11(5):1258-1264 2020(May)
Author:Yuji Shibata; Nobuaki Kobayashi; Takashi Sato; Kentaro Nakashima; Takeshi Kaneko


Oral Administration of Flavonifractor plautii Strongly Suppresses Th2 Immune Responses in Mice
Frontiers in Immunology,11 2020(Feb. 28)
Author:Tasuku Ogita; Yoshinari Yamamoto; Ayane Mikami; Suguru Shigemori; Takashi Sato; Takeshi Shimosato


Lactobacillus ingluviei C37 from chicken inhibits inflammation in LPS‐stimulated mouse macrophages
Animal Science Journal,91(1) 2020(Jan.)
Author:Masami Tsukagoshi; Merisa Sirisopapong; Fu Namai; Minori Ishida; Supattra Okrathok; Suguru Shigemori; Tasuku Ogita; Takashi Sato; Sutisa Khempaka; Takeshi Shimosato


Oral priming with oligodeoxynucleotide particles from Lactobacillus rhamnosus GG attenuates symptoms of dextran sodium sulfate‐induced acute colitis in mice
Animal Science Journal,91(1) 2020(Jan.)
Author:Suguru Shigemori; Fu Namai; Tasuku Ogita; Takashi Sato; Takeshi Shimosato


Hepcidin exerts a negative immunological effect in pulmonary tuberculosis without HIV co-infection, prolonging the time to culture-negative.
International journal of infectious diseases : IJID : official publication of the International Society for Infectious Diseases,86:47-54 2019(Sep.)
Author:Ken Tashiro; Masaki Yamamoto; Ryota Ushio; Nobuaki Kobayashi; Takashi Sato; Makoto Kudo; Takeshi Kaneko
Abstract:OBJECTIVES: A major regulatory peptide in iron metabolism, hepcidin, has been shown to predict mortality in HIV-infected tuberculosis patients. The aim of this study was to evaluate whether plasma hepcidin levels on admission can be used to predict the treatment outcome of patients with smear-positive pulmonary tuberculosis (PTB) without HIV co-infection. METHODS: In this prospective observational study, a total of 35 PTB patients with Mycobacterium tuberculosis-positive sputum smears were enrolled. The relationship between plasma hepcidin levels on admission and the time period to sputum culture-negative was explored. RESULTS: Plasma hepcidin levels of PTB patients were significantly higher than those of healthy subjects (p<0.001). A positive correlation between hepcidin level on admission and the period until culture-negative was also observed (r=0.46, p=0.006). Furthermore, the hepcidin level showed a negative correlation with spot numbers in the positive control wells of the T-SPOT.TB assay; thus the effect of the peptide on interferon-gamma production in T cells was explored. Hepcidin reduced interferon-gamma gene transcription and interferon-gamma production in a dose-dependent manner in Jurkat cells stimulated with phytohaemagglutinin, an antigen non-specific stimulation. CONCLUSIONS: These findings indicate that hepcidin alters immunological reactions against M. tuberculosis infection and has an influence on the outcomes of PTB patients without HIV co-infection.


Clinical significance of Charlson comorbidity index as a prognostic parameter for patients with acute or subacute idiopathic interstitial pneumonias and acute exacerbation of collagen vascular diseases-related interstitial pneumonia.
Journal of thoracic disease,11(6):2448-2457 2019(Jun.)
Author:Kota Murohashi; Yu Hara; Yusuke Saigusa; Nobuaki Kobayashi; Takashi Sato; Masaki Yamamoto; Makoto Kudo; Takeshi Kaneko
Abstract:Background: A prognostic factor for patients with acute or subacute idiopathic interstitial pneumonias (IIPs) or acute exacerbation (AE) of collagen vascular diseases-related interstitial pneumonia (CVD-IP) has not been established. We aimed to determine whether the Charlson comorbidity index (CCI) could serve as a prognostic factor for patients with these patients. Methods: We assessed baseline prognostic factors among patients with acute or subacute IIPs and AE of CVD-IP who were admitted to hospital between January 2014 and December 2017. We classified them as survivors and non-survivors at 3 months and compared their age, sex, CCI, blood parameters [lactate dehydrogenase (LDH), surfactant protein (SP)-D, Krebs von den Lungen-6, and partial pressure of oxygen in arterial blood/fraction of the inspiratory oxygen], high resolution CT (HRCT) scores and treatment. Results: Sixty eight patients with (mean age, 75 years), were assessed. All patients received steroid pulse therapy. We found that 45 of acute or subacute IIPs and 16 of AE of CVD-IP were included. Stepwise multivariate analysis selected CCI (OR, 1.306; 95% CI, 1.090-1.573; P=0.004), serum LDH (OR, 1.003; 95% CI, 1.001-1.005; P=0.002), and sex (OR, 8.555; 95% CI, 1.729-154.978; P=0.038) as significant predictors of 3-month mortality among these patients. Three-month mortality was significantly worse among patients with high (≥4) than low (<4) CCI (mortality rates: 63.2% vs. 16.3%, P<0.001). Moreover, the composite scoring system including CCI, serum LDH, and sex was acceptable (Bootstrap AUC, 0.859; Bootstrap C-index, 0.747). Conclusions: The composite scoring system including CCI, sex, and serum LDH could be a useful mortality prediction tool for patients with acute or subacute IIPs and AE of CVD-IP requiring steroid pulse therapy.


Recombinant mouse calcitonin gene-related peptide secreted by Lactococcus lactis inhibits lipopolysaccharide-induced inflammatory response in macrophages.
Animal science journal = Nihon chikusan Gakkaiho,89(12):1707-1711 2018(Dec.)
Author:Fu Namai; Yoshinari Yamamoto; Takashi Sato; Tasuku Ogita; Takeshi Shimosato
Abstract:We describe the development of a genetically modified strain of lactic acid bacteria (gmLAB) capable of producing a recombinant mouse calcitonin gene-related peptide (rCGRP). This strain (NZ-CGRP) was generated by introducing a CGRP secretion plasmid into Lactococcus lactis NZ9000. Western blotting confirmed the secretion of rCGRP in the presence of the inducer nisin. Highly purified rCGRP was obtained from the culture supernatants of NZ-CGRP. We demonstrated that prophylactic exposure of a culture of mouse peritoneal macrophages to rCGRP inhibited lipopolysaccharide (LPS) induction of tumor necrosis factor-α (TNF-α). The rCGRP-secreting gmLAB strain holds promise for development as a new anti-inflammatory prophylactic.


Silicosis and lung cancer: current perspectives
Lung Cancer (Auckl),9:91-101 2018(Oct.)
Author:Sato T; Shimosato T; Klinman DM


Regulatory role of heme oxygenase-1 in silica-induced lung injury
Respiratory Research,19(1) 2018(Aug. 01)
Author:Kentaro Nakashima; Takashi Sato; Suguru Shigemori; Takeshi Shimosato; Masaharu Shinkai; Takeshi Kaneko


Immune synergistic oligodeoxynucleotide from Lactobacillus rhamnosus GG enhances the immune response upon co-stimulation by bacterial and fungal cell wall components.
Anim Sci J,89(10):1504-1511 2018(Jul.)
Author:Nigar S; Yamamoto Y; Okajima T; Sato T; Ogita T; Shimosato T
Abstract:Bacterial genomic DNA has recently been shown to elicit a highly evolved immune defense. This response can be selectively triggered for a wide range of therapeutic applications, including use as a vaccine adjuvant to immunotherapies for allergy, cancer, and infectious diseases. Previously, we identified a low-concentration immune synergistic oligodeoxynucleotide (iSN-ODN, named iSN34) from Lactobacillus rhamnosus GG that has immunosynergistic activity upon costimulation of target cells with ligands of Toll-like receptor 9 (TLR9). Here, we extend that observation by demonstrating the synergistic induction (in mouse splenocytes) of IL-6 by the combination of iSN34 with cell wall components of bacteria and fungi. We observed that splenocytes pretreated with iSN34 and then costimulated with agonists for TLR1/2 (Pam3 CSK4 ), TLR4 (lipopolysaccharide), or TLR2/6 (Zymosan) exhibited enhanced accumulation of IL-6. These results suggested that the combination of iSN34 with TLR1/2, TLR4, or TLR2/6 agonists may permit the induction of a potent immune response.


Recombinant Mouse Osteocalcin Secreted by Lactococcus lactis Promotes Glucagon-Like Peptide-1 Induction in STC-1 Cells.
Current microbiology,75(1):92-98 2018(Jan.)
Author:Fu Namai; Suguru Shigemori; Koichi Sudo; Takashi Sato; Yoshinari Yamamoto; Shireen Nigar; Tasuku Ogita; Takeshi Shimosato
Abstract:An osteoblastic protein, osteocalcin (OC), exists in vivo in two forms: carboxylated OC, and uncarboxylated or low-carboxylated OC (ucOC). ucOC acts as a hormone to regulate carbon and energy metabolism. Recent studies demonstrated that ucOC exerts insulinotropic effects, mainly through the glucagon-like peptide 1 (GLP-1) pathway. GLP-1 is an insulinotropic hormone secreted by enteroendocrine L cells in the small intestine. Thus, efficient delivery of ucOC to the small intestine may be a new therapeutic option for metabolic diseases such as diabetes and obesity. Here, we genetically engineered a lactic acid bacterium, Lactococcus lactis, to produce recombinant mouse ucOC. Western blotting showed that the engineered strain (designated NZ-OC) produces and secretes the designed peptide (rOC) in the presence of nisin, an inducer of the recombinant gene. Highly-purified rOC was obtained from the culture supernatants of NZ-OC using immobilized metal affinity chromatography. An in vitro assay showed that purified rOC promotes GLP-1 secretion in a mouse intestinal neuroendocrine cell line, STC-1, in a dose-dependent manner. These results clearly demonstrate that NZ-OC secretes rOC, and that rOC can promote GLP-1 secretion by STC-1 cells. Genetically modified lactic acid bacteria (gmLAB) have been proposed over the last two decades as an effective and low-cost mucosal delivery vehicle for biomedical proteins. NZ-OC may be an attractive tool for the delivery of rOC to trigger GLP-1 secretion in the small intestine to treat diabetes and obesity.


The platelet count can predict in-hospital death in hiv-negative smear-positive pulmonary tuberculosis inpatients
Internal Medicine,57(10):1391-1397 2018
Author:Hideto Goto; Nobuyuki Horita; Ken Tashiro; Kenjiro Nagai; Masaki Yamamoto; Takashi Sato; Yu Hara; Hideyuki Nagakura; Yuji Shibata; Hiroki Watanabe; Kentaro Nakashima; Ryota Ushio; Akimichi Nagashima; Misako Ikeda; Atsuya Narita; Katsuhito Sasaki; Nobuaki Kobayashi; Makoto Kudo; Takeshi Kaneko
Abstract:Objective This retrospective cohort study investigated whether the three components of the blood cell count have prognostic implications in HIV-negative Japanese adult inpatients with smear-positive pulmonary tuberculosis. Methods We reviewed patients who were treated by the isoniazid, rifampicin, pyrazinamide, and ethambutol regimen or by the isoniazid, rifampicin, and ethambutol regimen. The association between the patient data on admission and the survival outcome was evaluated. Results We reviewed 367 consecutive patients (male, 60.5%) with a median age of 72 [interquartile range (IQR), 54-82] years. While the white blood cell count did not differ between the two groups, (discharged alive: 7,000/μL IQR, 5,500-9,300 died in hospital: 7,200/μL IQR, 5,600-9,400 p=0.797), hemoglobin level (discharged alive: 11.5 g/dL IQR, 10.0-13.1 died in hospital: 9.9 g/dL IQR, 8.6-11.3 p< 0.001) and the platelet count (discharged alive: 275,000/μL IQR, 206,000-345,000 died in hospital: 149,000/μL IQR, 93,000-236,000 p< 0.001) were lower in patients who died in hospital. After dividing patients into hemoglobin- and platelet-based quantiles, the lower quantile class tended to show poorer survival (log-rank test for trend p< 0.001 for both). A multi-variable Cox proportional hazards model revealed that hazard ratio for in-hospital death for every 1,000/μL increase of platelet count was 0.997 (95%CI, 0.995-0.999 p=0.010) the hazard ratio for the hemoglobin level was not significant. Conclusion A low platelet count was clearly related to a poor life prognosis in HIV-negative Japanese adult inpatients with smear-positive pulmonary tuberculosis.


Synergistic oligodeoxynucleotide strongly promotes CpG-induced interleukin-6 production
BMC IMMUNOLOGY,18:44 2017(Oct.)
Author:Shireen Nigar; Yoshinari Yamamoto; Takuma Okajima; Suguru Shigemori; Takashi Sato; Tasuku Ogita; Takeshi Shimosato
Abstract:Background: Bacterial genomes span a significant portion of diversity, reflecting their adaptation strategies; these strategies include nucleotide usage biases that affect chromosome configuration. Here, we explore an immuno-synergistic oligodeoxynucleotide (iSN-ODN, named iSN34), derived from Lactobacillus rhamnosus GG (LGG) genomic sequences, that exhibits a synergistic effect on immune response to CpG-induced immune activation. Methods: The sequence of iSN34 was designed based on the genomic sequences of LGG. Pathogen-free mice were purchased from Japan SLC and maintained under temperature- and light-controlled conditions. We tested the effects of iSN34 exposure in vitro and in vivo by assessing effects on mRNA expression, protein levels, and cell type in murine splenocytes. Results: We demonstrate that iSN34 has a significant stimulatory effect when administered in combination with CpG ODN, yielding enhanced interleukin (IL)-6 expression and production. IL-6 is a pleotropic cytokine that has been shown to prevent epithelial apoptosis during prolonged inflammation. Conclusions: Our results are the first report of a bacterial-DNA-derived ODN that exhibits immune synergistic activity. The potent over-expression of IL-6 in response to treatment with the combination of CpG ODN and iSN34 suggests a new approach to immune therapy. This finding may lead to novel clinical strategies for the prevention or treatment of dysfunctions of the innate and adaptive immune systems.


Genetically modified Lactococcus lactis producing a green fluorescent protein bovine lactoferrin fusion protein suppresses proinflammatory cytokine expression in lipopolysaccharide-stimulated RAW 264.7 cells
JOURNAL OF DAIRY SCIENCE,100(9):7007-7015 2017(Sep.)
Author:S. Shigemori; F. Namai; Y. Yamamoto; S. Nigar; T. Sato; T. Ogita; T. Shimosato
Abstract:Lactoferrin (LF), an iron-binding glycoprotein distributed widely in the biological fluids of mammals, is believed to play an important role in host defenses against infection. Previous studies in animal models and humans demonstrated that combined administration of LF and probiotic lactic acid bacteria (LAB) can prevent sepsis. In this study, we genetically engineered a probiotic LAB strain, Lactococcus lactis, to produce recombinant bovine LF based on the green fluorescent protein (GFP)-fused expression system. Western blotting confirmed that the genetically modified L. lactis strain (designated NZ-GFP-bLF) produced a protein corresponding to a fusion of GFP and bLF in the presence of nisin, an inducer of target gene expression. The protein synthesized by NZ-GFP-bLF was fluorescent and thus we monitored the time-dependent change in the production level of the recombinant protein using fluorometric analysis. The utility of NZ-GFP-bLF in preventing sepsis was determined by investigating its anti-inflammatory property in lipopolysaccharide (LPS)-stimulated mouse macrophage RAW 264.7 cells. Pretreatment of RAW 264.7 cells with NZ-GFP-bLF significantly attenuated the LPS-induced mRNA expression and protein production of 3 proinflammatory cytokines (IL-l alpha, IL-6, and tumor necrosis factor-alpha) compared with pretreatment with a vector control strain of L. lactis. Our results suggest that NZ-GFP-bLF holds promise for the development of a new prophylaxis for sepsis.


Class A CpG Oligonucleotide Priming Rescues Mice from Septic Shock via Activation of Platelet-Activating Factor Acetylhydrolase
FRONTIERS IN IMMUNOLOGY,8:1049 2017(Aug.)
Author:Yoshinari Yamamoto; Ryu Sugimura; Takafumi Watanabe; Suguru Shigemori; Takuma Okajima; Shireen Nigar; Fu Namai; Takashi Sato; Tasuku Ogita; Takeshi Shimosato
Abstract:Sepsis is a life-threatening, overwhelming immune response to infection with high morbidity and mortality. Inflammatory response and blood clotting are caused by sepsis, which induces serious organ damage and death from shock. As a mechanism of pathogenesis, platelet-activating factor (PAF) induces excessive inflammatory responses and blood clotting. In this study, we demonstrate that a Class A CpG oligodeoxynucleotide (CpG-A(1585)) strongly induced PAF acetylhydrolase, which generates lyso-PAF. CpG-A(1585) rescued mice from acute lethal shock and decreased fibrin deposition, a hallmark of PAF-induced disseminated intravascular coagulation. Furthermore, CpG-A(1585) improved endotoxin shock induced by lipopolysaccharide, which comprises the cell wall of Gram-negative bacteria and inhibits inflammatory responses induced by cytokines such as interleukin-6 and tumor necrosis factor-alpha. These results suggest that CpG-A(1585) is a potential therapeutic target to prevent sepsis-related induction of PAF.


HbA1c level cannot predict the treatment outcome of smearpositive non-multi-drug-resistant HIV-negative pulmonary tuberculosis inpatients
SCIENTIFIC REPORTS,7:46488 2017(Apr.)
Author:Ken Tashiro; Nobuyuki Horita; Kenjiro Nagai; Misako Ikeda; Masaharu Shinkai; Masaki Yamamoto; Takashi Sato; Yu Hara; Hideyuki Nagakura; Yuji Shibata; Hiroki Watanabe; Kentaro Nakashima; Ryota Ushio; Akimichi Nagashima; Atsuya Narita; Nobuaki Kobayashi; Makoto Kudo; Takeshi Kaneko
Abstract:We conducted a single-center retrospective cohort study to evaluate whether the HbA1c level on admission could predict the in-hospital treatment outcome of smear-positive non-multi-drug-resistant HIV-negative culture-proven pulmonary tuberculosis inpatients. Our standard regimens under the direct observation were HRZE or HRE for the first two months followed by combination therapy with isoniazid and rifampicin. Our cohort consisted of consecutive 239 patients consisted of 147 men and 92 women with a median age of 73 years. The HbA1c level of patients whose HbA1c was above 7.0% on admission showed clear declining trends after admission. HbA1c on admission had no Spearman's rank correlation with time to discharge alive (r = 0.17) and time to becoming non-infective (r = 0.17). By Kaplan-Meier curves and a log-rank trend test, HbA1c quartile subgroups showed no association with times to discharge alive (p = 0.431), becoming non-infective (p = 0.113), and in-hospital death (p = 0.427). Based on multi-variate Cox analysis, HbA1c on admission had no significant impact on time to discharge alive (hazard ratio = 1.03, 95% CI 0.89-1.20, p = 0.659), becoming non-infective (hazard ratio = 0.93, 95% CI 0.80-1.06, p = 0.277), and in-hospital death (hazard ratio = 0.68, 0.43-1.07, p = 0.097). In conclusion, the HbA1c level on admission did not seem to affect in-hospital tuberculosis treatment outcomes in Japanese cohort.


Secretion of an immunoreactive single-chain variable fragment antibody against mouse interleukin 6 by Lactococcus lactis
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,101(1):341-349 2017(Jan.)
Author:Suguru Shigemori; Masaki Ihara; Takashi Sato; Yoshinari Yamamoto; Shireen Nigar; Tasuku Ogita; Takeshi Shimosato
Abstract:Interleukin 6 (IL-6) is an important pathogenic factor in development of various inflammatory and autoimmune diseases and cancer. Blocking antibodies against molecules associated with IL-6/IL-6 receptor signaling are an attractive candidate for the prevention or therapy of these diseases. In this study, we developed a genetically modified strain of Lactococcus lactis secreting a single-chain variable fragment antibody against mouse IL-6 (IL6scFv). An IL6scFv-secretion vector was constructed by cloning an IL6scFv gene fragment into a lactococcal secretion plasmid and was electroporated into L. lactis NZ9000 (NZ-IL6scFv). Secretion of recombinant IL6scFv (rIL6scFv) by nisin-induced NZ-IL6scFv was confirmed by western blotting and was optimized by tuning culture conditions. We found that rIL6scFv could bind to commercial recombinant mouse IL-6. This result clearly demonstrated the immunoreactivity of rIL6scFv. This is the first study to engineer a genetically modified strain of lactic acid bacteria (gmLAB) that produces a functional anti-cytokine scFv. Numerous previous studies suggested that mucosal delivery of biomedical proteins using gmLAB is an effective and low-cost way to treat various disorders. Therefore, NZ-IL6scFv may be an attractive tool for the research and development of new IL-6 targeting agents for various inflammatory and autoimmune diseases as well as for cancer.


Factors for Predicting Outcomes among Non-HIV Patients with Pulmonary Tuberculosis
INTERNAL MEDICINE,56(24):3277-3282 2017
Author:Toshinori Tsukahara; Nobuyuki Horita; Ken Tashiro; Kenjiro Nagai; Masaharu Shinkai; Masaki Yamamoto; Takashi Sato; Yu Hara; Hideyuki Nagakura; Yuji Shibata; Hiroki Watanabe; Kentaro Nakashima; Ryota Ushio; Akimichi Nagashima; Misako Ikeda; Atsuya Narita; Katsuhito Sasaki; Nobuaki Kobayashi; Makoto Kudo; Takeshi Kaneko
Abstract:Objective Onodera's Prognostic Nutritional Index (PNI), determined as "10x albumin (g/dL) + 0.005x lymphocyte count (/mu L)," was originally designed to determine the risk of complications following gastrointestinal surgery. This single-center, retrospective observational study was designed to investigate whether or not the PNI can predict the treatment outcome. Methods We consecutively reviewed HIV-negative pulmonary tuberculosis adults in an isolation ward. Most patients were being treated with standard three-or four-drug regimens. Patients were discharged after consecutive negative smears/cultures were confirmed. The risk of all-cause death was assessed using a multivariable Cox proportional hazard model and a log-rank trend test. Results During the observation period, we observed 371 consecutive patients with a median age of 72 (interquartile range [IQR]: 54-82) years. In our cohort, 295 (79.5%) patients were discharged alive, and 76 (20.5%) died in-hospital. Patients who died in-hospital had a lower PNI [median 21.2 (IQR: 18.5-25.9)] than those who were discharged alive [median 35.1 (IQR: 28.0-43.3); p<0.001]. The area under the receiver operating characteristic curve was 0.87. After dividing the patients based on the baseline PNI quartile, those patients with a lower PNI showed a poorer survival than those with a higher PNI (log-rank trend p<0.001). After adjusting for other baseline variables, the baseline PNI was still associated with in-hospital death with a hazard ratio of 0.86 (95% confidence interval: 0.82-0.91, p<0.001). Conclusion Our results showed that a low PNI was clearly related to a poor survival prognosis in smearpositive HIV-negative pulmonary tuberculosis inpatients.


Diagnostic test accuracy of loop-mediated isothermal amplification assay for Mycobacterium tuberculosis: systematic review and meta-analysis
SCIENTIFIC REPORTS,6:39090 2016(Dec.)
Author:Kenjiro Nagai; Nobuyuki Horita; Masaki Yamamoto; Toshinori Tsukahara; Hideyuki Nagakura; Ken Tashiro; Yuji Shibata; Hiroki Watanabe; Kentaro Nakashima; Ryota Ushio; Misako Ikeda; Atsuya Narita; Akinori Kanai; Takashi Sato; Takeshi Kaneko
Abstract:Diagnostic test accuracy of the loop-mediated isothermal amplification (LAMP) assay for culture proven tuberculosis is unclear. We searched electronic databases for both cohort and case-control studies that provided data to calculate sensitivity and specificity. The index test was any LAMP assay including both commercialized kits and in-house assays. Culture-proven M. tuberculosis was considered a positive reference test. We included 26 studies on 9330 sputum samples and one study on 315 extra-pulmonary specimens. For sputum samples, 26 studies yielded the summary estimates of sensitivity of 89.6% (95% CI 85.6-92.6%), specificity of 94.0% (95% CI 91.0-96.1%), and a diagnostic odds ratio of 145 (95% CI 93-226). Nine studies focusing on Loopamp MTBC yielded the summary estimates of sensitivity of 80.9% (95% CI 76.0-85.1%) and specificity of 96.5% (95% CI 94.7-97.7%). Loopamp MTBC had higher sensitivity and lower specificity for smear-positive sputa compared to smear-negative sputa. In-house assays showed higher sensitivity and lower specificity compared to Loopamp MTBC. LAMP promises to be a useful test for the diagnosis of TB, however there is still need to improve the assay to make it simpler, cheaper and more efficient to make it competitive against other PCR methods already available.


Clarithromycin Suppresses Chloride Channel Accessory 1 and Inhibits Interleukin-13-Induced Goblet Cell Hyperplasia in Human Bronchial Epithelial Cells
ANTIMICROBIAL AGENTS AND CHEMOTHERAPY,60(11):6585-6590 2016(Nov.)
Author:Akimichi Nagashima; Masaharu Shinkai; Masahiro Shinoda; Tadasuke Shimokawaji; Yasuhiro Kimura; Kei Mishina; Takashi Sato; Mariko Toda; Yoshiaki Inayama; Bruce K. Rubin; Takeshi Kaneko
Abstract:Activation of the interleukin-13 (IL-13) receptor leads to signal transducer and activator of transcription 6 (STAT6) activation and subsequent induction of SAM pointed domain containing ETS transcription factor (SPDEF) and chloride channel accessory 1 (CLCA1), increasing secretion of the gel-forming mucin MUC5AC. Activation of the epidermal growth factor receptor (EGFR) also leads to MUC5AC production via extracellular signal-regulated kinase (ERK1/2). We examined the effect of clarithromycin IL-13 signaling leading to production. Normal human bronchial epithelial (NHBE) cells were grown for 14 days at an air-liquid interface (ALI) with IL-13 and/or clarithromycin. Histochemical analysis was performed using hematoxylin and eosin (HE) staining and MUC5AC immunostaining. MUC5AC, SPDEF, and CLCA1 mRNA expression were evaluated by real-time PCR. Western analysis was used to assess phosphorylation of STAT6 and ERK1/2. Clarithromycin decreased IL-13-induced goblet cell hyperplasia and MUC5AC mRNA expression in a dose-dependent manner. Clarithromycin decreased IL-13-stimulated SPDEF and CLCA1 mRNA expression in a dose-dependent manner, and at 32 mu g/ml CLCA1 was profoundly decreased (P < 0.001). Although clarithromycin had no effect on STAT6 phosphorylation induced by IL-13, it decreased constitutive phosphorylation of ERK1/2 (P< 0.05).


Use of nanoparticles to deliver immunomodulatory oligonucleotides
WILEY INTERDISCIPLINARY REVIEWS-NANOMEDICINE AND NANOBIOTECHNOLOGY,8(4):631-637 2016(Jul.)
Author:Dennis M. Klinman; Takashi Sato; Takeshi Shimosato
Abstract:Synthetic oligonucleotides (ODNs) containing unmethylated 'CpG motifs' stimulate the innate immune system to produce cytokines, chemokines, and polyreactive antibodies. CpG ODNs have shown promise as vaccine adjuvants and for the treatment of infectious diseases and cancer. The immunostimulatory activity of CpG ODNs is inhibited by DNA-containing 'suppressive' motifs. ODNs expressing suppressive motifs (Sup ODNs) reduce ongoing immune reactions and show promise in the treatment of autoimmune and inflammatory diseases. This work reviews recent progress in the use of nanoparticles as carriers of CpG and Sup ODNs to target their delivery to the GI tract and lungs. (C) 2015 Wiley Periodicals, Inc.


Digital PCR assay detection of circulating Mycobacterium tuberculosis DNA in pulmonary tuberculosis patient plasma
TUBERCULOSIS,99:47-53 2016(Jul.)
Author:Ryota Ushio; Masaki Yamamoto; Kentaro Nakashima; Hiroki Watanabe; Kenjiro Nagai; Yuji Shibata; Ken Tashiro; Toshinori Tsukahara; Hideyuki Nagakura; Nobuyuki Horita; Takashi Sato; Masaharu Shinkai; Makoto Kudo; Atsuhisa Ueda; Takeshi Kaneko
Abstract:Nucleic acid amplification tests are a major diagnostic tool for pulmonary tuberculosis (PTB). Recently, digital PCR (dPCR) assay has improved sensitivity for the detection of small copy numbers of target molecules. The aim of this study is to explore the utility of dPCR for detecting Mycobacterium tuberculosis (MTB) DNA in PTB patient plasma. Total DNA was purified from plasma samples of newly diagnosed sputum smear-positive PTB patients. Copy numbers of MTB-specific genes in the samples were quantified with dPCR assays targeted for IS6110 or gyrB. A total of 33 PTB patients were enrolled. Significant differences between PTB patients and controls were observed in copy numbers of both targets: IS6110 mean +/- SD, 144.8 +/- 538.3 vs 0.44 +/- 0.49 (copies/20 mu L, p = 0.004; Manne-Whitney U test) and gyrB mean +/- SD, 359.0 +/- 2116 vs 0.07 +/- 0.28 (copies/20 mu L, p = 0.011; Manne-Whitney U test), respectively. This test had sensitivities of 65% or 29% and a specificity of 93% or 100% with the IS6110-targeted or gyrB-targeted assays, respectively. A dPCR assay successfully detected MTB DNA in PTB patient plasma. This minimally invasive and accurate method has potential to become an alternative diagnostic option. (C) 2016 Elsevier Ltd. All rights reserved.


Diagnostic test accuracy of anti-glycopeptidolipid-core IgA antibodies for Mycobacterium avium complex pulmonary disease: systematic review and meta-analysis
SCIENTIFIC REPORTS,6:29325 2016(Jul.)
Author:Yuji Shibata; Nobuyuki Horita; Masaki Yamamoto; Toshinori Tsukahara; Hideyuki Nagakura; Ken Tashiro; Hiroki Watanabe; Kenjiro Nagai; Kentaro Nakashima; Ryota Ushio; Misako Ikeda; Atsuya Narita; Akinori Kanai; Takashi Sato; Takeshi Kaneko
Abstract:Currently, an anti-glycopeptidolipid (GPL)-core IgA antibody assay kit for diagnosing Mycobacterium avium complex (MAC) is commercially available. We conducted this systematic review and meta-analysis to reveal the precise diagnostic accuracy of anti-GPL-core IgA antibodies for MAC pulmonary disease (MAC-PD). We systematically searched reports that could provide data for both sensitivity and specificity by anti-GPL-core IgA antibody for clinically diagnosed MAC-PD. Diagnostic test accuracy was estimated using the bivariate model. Of the 257 articles that we had found through primary search, we finally included 16 reports consisted of 1098 reference positive subjects and 2270 reference negative subjects. The diagnostic odds ratio was 24.8 (95% CI 11.6-52.8, I-2 = 5.5%) and the area under the hierarchical summary receiver operating characteristic curves was 0.873 (95% CI 0.837-0.913). With a cutoff value of 0.7 U/mL, the summary estimates of sensitivity and specificity were 0.696 (95% CI 0.621-0.761) and 0.906 (95% CI 0.836-0.951), respectively. The positive and negative likelihood ratios were 7.4 (95% CI 4.1-13.8) and 0.34 (95% CI 0.26-0.43), respectively. The demanding clinical diagnostic criteria may be a cause of false positive of the index test. The index test had good overall diagnostic accuracy and was useful to ruling in MAC-PD with the cutoff value.


Development of a simple IgE-independent anaphylactic model using buckwheat antigen and B-type CpG oligodeoxynucleotide from Streptococcus thermophilus
ANIMAL SCIENCE JOURNAL,87(5):710-717 2016(May)
Author:Yoshinari Yamamoto; Suguru Shigemori; Shireen Nigar; Kazushi Oshiro; Yeqin Wang; Takashi Sato; Takeshi Shimosato
Abstract:We developed a severe anaphylactic model in mice using buckwheat antigen and B-type CpG-oligodeoxynucleotides (CpG-ODNs) from Streptococcus thermophilus genome. In typical systemic anaphylaxis models, animals are challenged with large quantity of antigens via an intravenous (i.v.) route. Here, we showed a simple anaphylactic shock after challenge via intraperitoneal (i.p.) route. The i.p. method is simpler than i.v. administration and has a lower risk for failure. To generate this anaphylactic model, 5-week-old female BALB/c mice were first i.p. sensitized with buckwheat antigen mixed with B-type CpG-ODN. After 2 weeks, mice were challenged with antigen to induce anaphylactic shock, which was evaluated by scoring the severity symptoms and measuring serum levels of various proteins and splenic cell producing cytokines. Immunoglobulin (Ig)G(2a) production and interferon-gamma positive cells were markedly increased in mice immunized with antigen mixed with B-type CpG-ODN, whereas serum IgE levels were decreased by B-type CpG-ODN. We also examined the effects of various ODNs (A, B and C-type CpG-ODNs) and antigens (buckwheat, alpha-casein, beta-lactoglobulin and ovalbumin) on anaphylactic severity, and found that the combination of buckwheat and B-type CpG-ODN induced the most intense anaphylactic shock. This model is expected to contribute to the study of the prevention of anaphylactic shock.


Multiple nodular pulmonary metastases of lung adenocarcinoma with epidermal growth factor receptor mutation
Respiratory Investigation,54(2):133-135 2016(Mar. 01)
Author:Natsuki Kawata; Miyo Inoue; Nobuyuki Horita; Koji Tomaru; Masaki Yamamoto; Takashi Sato; Naoki Miyazawa; Takeshi Kaneko


Age, Dehydration, Respiratory Failure, Orientation Disturbance, and Blood Pressure Score Predicts In-hospital Mortality in HIV-negative Non-multidrug-resistant Smear-positive Pulmonary Tuberculosis in Japan
Scientific Reports,6:21610 2016(Feb. 17)
Author:Kenjiro Nagai; Nobuyuki Horita; Takashi Sato; Masaki Yamamoto; Hideyuki Nagakura; Takeshi Kaneko
Abstract:The A-DROP scoring system was originally designed to assess clinical severity of community acquired pneumonia using the following parameters: advanced Age, Dehydration, Respiratory failure, Orientation disturbance (confusion) and, low blood Pressure. Total A-DROP score ranges zero to five assigning one point for each component, wherein five indicates the poorest prognosis. The purpose of this single-center retrospective study was to determine whether A-DROP could predict the risk for death in patients with pulmonary tuberculosis. We reviewed consecutive HIV-negative, non-multidrug-resistant smear-positive adult pulmonary tuberculosis patients. The cohort consisted of 134 men (38.8%), 211 women (61.2%), 272 who discharged alive (28.8%), and 73 who died in-hospital (21.2%) with a median age of 72 (IQR: 54-82) years. A one-point increase in the A-DROP score was associated with a higher risk for in-hospital mortality with odds ratio of 3.8 (95% confidence interval 2.8-5.2, P < 0.001). The area under receiver operating characteristics curve was 0.86. The total score cutoff of 1.5 provided the best Youden Index of 0.61. Using this criteria, total score > 1.5, sensitivity was 85% and specificity was 76%. Kaplan-Meier curve clearly indicated that in-hospital mortality increased with higher A-DROP scores (Log-rank test < 0.001). In conclusion, A-DROP score clearly indicate pulmonary tuberculosis in-hospital mortality.


Amrubicin for relapsed small-cell lung cancer: a systematic review and meta-analysis of 803 patients
SCIENTIFIC REPORTS,6 2016(Jan.)
Author:Nobuyuki Horita; Masaki Yamamoto; Takashi Sato; Toshinori Tsukahara; Hideyuki Nagakura; Ken Tashiro; Yuji Shibata; Hiroki Watanabe; Kenjiro Nagai; Kentaro Nakashima; Ryota Ushio; Misako Ikeda; Nobuaki Kobayashi; Masaharu Shinkai; Makoto Kudo; Takeshi Kaneko
Abstract:Currently, amrubicin is permitted for relapsed small-cell lung carcinoma (SCLC) only in Japan. The efficacy and adverse effects of amrubicin as reported by previous studies varied greatly. The inclusion criterion was a prospective study that was able to provide data for efficacy and safety by the AMR single agent regimen as second-line chemotherapy for a patient with SCLC. Binary data were meta-analyzed with the random-model generic inverse variance method. We included nine articles consisted of 803 patients. The pooled three-, six-, and nine-month progression-free survival were 63% (95% CI 57-69%, I-2 = 53%), 28% (95% CI 21-35%, I-2 = 71%), and 10% (95% CI 6-14%, I-2 = 41%), respectively. The pooled six-, 12-, and 18-month overall survival were 69% (95% CI 61-78%, I-2 = 83%), 36% (95% CI 28-44%, I-2 = 80%), and 15% (95% CI 8-21%, I-2 = 81%), respectively. Amrubicin seemed much more beneficial for Japanese patients. However, compared to the efficacy of topotecan presented in a previous meta-analysis, amrubicin may be a better treatment option than topotecan for both Japanese and Euro-American. Adverse effects by amrubicin were almost exclusively observed to be hematological. Notably, grade III/IV neutropenia incidence was 70% and febrile neutropenia incidence was 12%.


Sensitivity and specificity of Cobas TaqMan MTB real-time polymerase chain reaction for culture-proven Mycobacterium tuberculosis: meta-analysis of 26999 specimens from 17 Studies.
Scientific reports,5:18113-18113 2015(Dec. 09)
Author:Nobuyuki Horita; Masaki Yamamoto; Takashi Sato; Toshinori Tsukahara; Hideyuki Nagakura; Ken Tashiro; Yuji Shibata; Hiroki Watanabe; Kenjiro Nagai; Kentaro Nakashima; Ryota Ushio; Misako Ikeda; Kentaro Sakamaki; Takashi Yoshiyama; Takeshi Kaneko
Abstract:Since 2010, studies on the diagnostic accuracy of COBAS TaqMan MTB (CTM) have been frequently reported with an unignorable discrepancy. The key inclusion criterion for this systematic review was original studies that could provide sufficient data for calculating the sensitivity and the specificity of CTM for M tuberculosis (TB) or M tuberculosis complex. The reference test was Mycobacterium culture. We used bivariate model for meta-analyses. Of the 201 candidate articles, we finally identified 17 eligible articles.Concerning the respiratory specimens, 1900 culture positive specimens and 20983 culture negative specimens from 15 studies were assessed. This provided the summary estimate sensitivity of 0.808 (95% CI 0.758-0.850) and the summary estimate specificity of 0.990 (95% CI 0.981-0.994). The area under curve was 0.956. The diagnostic odds ratio was 459 (95% CI 261-805, I(2) 26%). For the smear positive respiratory specimens, the sensitivity was 0.952 (95% CI 0.926-0.969) and the specificity was 0.916 (95% CI 0.797-0.968). For the smear negative respiratory specimens, the sensitivity and the specificity were 0.600 (95% CI 0.459-0.726) and 0.989 (95% CI 0.981-0.993), respectively. The diagnostic accuracy was poorer for the non-respiratory specimens, than for the respiratory specimens, but was acceptable. We believe that the information obtained from this study will aid physicians' decision making.


Oral delivery of Lactococcus lactis that secretes bioactive heme oxygenase-1 alleviates development of acute colitis in mice
MICROBIAL CELL FACTORIES,14(1):189 2015(Nov.)
Author:Suguru Shigemori; Takafumi Watanabe; Kai Kudoh; Masaki Ihara; Shireen Nigar; Yoshinari Yamamoto; Yoshihito Suda; Takashi Sato; Haruki Kitazawa; Takeshi Shimosato
Abstract:Background: Mucosal delivery of therapeutic proteins using genetically modified strains of lactic acid bacteria (gmLAB) is being investigated as a new therapeutic strategy. Methods: We developed a strain of gmLAB, Lactococcus lactis NZ9000 (NZ-HO), which secretes the anti-inflammatory molecule recombinant mouse heme oxygenase-1 (rmHO-1). The effects of short-term continuous oral dosing with NZ-HO were evaluated in mice with dextran sulfate sodium (DSS)-induced acute colitis as a model of inflammatory bowel diseases (IBD). Results: We identified the secretion of rmHO-1 by NZ-HO. rmHO-1 was biologically active as determined with spectroscopy. Viable NZ-HO was directly delivered to the colon via oral administration, and rmHO-1 was secreted onto the colonic mucosa in mice. Acute colitis in mice was induced by free drinking of 3 % DSS in water and was accompanied by an increase in the disease activity index score and histopathological changes. Daily oral administration of NZ-HO significantly improved these colitis-associated symptoms. In addition, NZ-HO significantly increased production of the anti-inflammatory cytokine interleukin (IL)-10 and decreased the expression of pro-inflammatory cytokines such as IL-1 alpha and IL-6 in the colon compared to a vector control strain. Conclusions: Oral administration of NZ-HO alleviates DSS-induced acute colitis in mice. Our results suggest that NZ-HO may be a useful mucosal therapeutic agent for treating IBD.


Intrapulmonary Delivery of CpG Microparticles Eliminates Lung Tumors
MOLECULAR CANCER THERAPEUTICS,14(10):2198-2205 2015(Oct.)
Author:Takashi Sato; Takeshi Shimosato; Atsuhisa Ueda; Yoshiaki Ishigatsubo; Dennis M. Klinman
Abstract:CpG oligonucleotides (ODN) stimulate the innate immune system by triggering cells that express TLR9. The resulting response promotes tumor regression, an effect optimized by delivery of CpG ODN to the tumor site. This work examines the effect of instilling CpG ODN adsorbed onto polyketal microparticles (CpG-MP) into the lungs of mice with non-small cell lung cancer. Intrapulmonary delivery of CpG-MP improved ODN uptake and retention at the tumor site, thereby inducing a stronger Th1 response than systemically administered or unadsorbed CpG ODN. CpG-MP reversed the immunosuppression that characterized the tumor microenvironment by (i) decreasing the number of immunosuppressive Tregs and M2 macrophages while (ii) increasing the number of tumoricidal CD8(+) T cells and M1 macrophages. These effects promoted tumor regression and culminated in 82% permanent survival of mice with otherwise fatal Lewis lung cancer. (C) 2015 AACR.


Topotecan for Relapsed Small-cell Lung Cancer: Systematic Review and Meta-Analysis of 1347 Patients
SCIENTIFIC REPORTS,5:15437 2015(Oct.)
Author:Nobuyuki Horita; Masaki Yamamoto; Takashi Sato; Toshinori Tsukahara; Hideyuki Nagakura; Ken Tashiro; Yuji Shibata; Hiroki Watanabe; Kenjiro Nagai; Miyo Inoue; Kentaro Nakashima; Ryota Ushio; Masaharu Shinkai; Makoto Kudo; Takeshi Kaneko
Abstract:Topotecan is the most reliable chemotherapy regimen for relapsed small-cell lung carcinoma (SCLC). The efficacy and adverse effects of topotecan as reported by previous studies varied greatly. The inclusion criterion was a prospective study that was able to provide data for 6-month over-all survival (OS) rate, 1-year OS rate, objective responses, and/or adverse effects of single agent topotecan as a second line chemotherapy for SCLC, written in English language as a full article. Any topotecan regimen were allowed. Binary data were meta-analyzed with the random-model generic inverse variance method. We included 14 articles consisted of 1347 patients. Pooled values were estimated as follows. <Refractory relapse> Six-month OS rate: 37% (95% CI: 28-46%). One-year OS rate: 9% (95% CI: 5-13%). Response rate: 5% (95% CI: 1-8%). <Sensitive relapse> Six-month OS rate: 57% (95% CI: 50-64%). One-year OS rate: 27% (95% CI: 22-32%). Response rate: 17% (95% CI: 11-23%). <Adverse effect> Grade III/IV neutropenia 69% (95% CI: 58-80%). Grade III/IV thrombopenia 41% (95% CI: 34-48%). Grade III/IV anemia 24% (95% CI: 17-30%). Non-hematorogical events were rare. Chemotherapy-related death 2% (95% CI: 1-3%). In conclusion, Topotecan provided a possibly promising outcome for sensitive-relapse SCLC and poor outcome for refractory relapse SCLC. Adverse events were mainly hematological.


Inhibitory/Suppressive Oligodeoxynucleotide Nanocapsules as Simple Oral Delivery Devices for Preventing Atopic Dermatitis in Mice
MOLECULAR THERAPY,23(2):297-309 2015(Feb.)
Author:Yeqin Wang; Yoshinari Yamamoto; Suguru Shigemori; Takafumi Watanabe; Kazushi Oshiro; Xinyu Wang; Pengfei Wang; Takashi Sato; Shinichi Yonekura; Sachi Tanaka; Haruki Kitazawa; Takeshi Shimosato
Abstract:Here, we report a simple and low-cost oral oligodeoxynucleotide (ODN) delivery system targeted to the gut Peyer's patches (PPs). This system requires only Dulbecco's modified eagle's medium, calcium chloride, ODNs, and basic laboratory equipment. ODN nanocapsules (ODNcaps) were directly delivered to the PPs through oral administration and were taken up by macrophages in the PPs, where they induced an immune response. Long-term continuous oral dosing with inhibitory/suppressive ODNcaps (iODNcaps, "iSG3caps" in this study) was evaluated using an atopic dermatitis mouse model to visually monitor disease course. Administration of iSG3caps improved skin lesions and decreased epidermal thickness. Underlying this effect is the ability of iSG3 to bind to and prevent phosphorylation of signal transducer and activator of transcription 6, thereby blocking the interleukin-4 signaling cascade mediated by binding of allergens to type 2 helper T cells. The results of our iSG3cap oral delivery experiments suggest that iSG3 may be useful for treating allergic diseases.


The effectiveness of sputum pH analysis in the prediction of response to therapy in patients with pulmonary tuberculosis.
PeerJ,3:e1448 2015
Author:Makoto Masuda; Takashi Sato; Kentaro Sakamaki; Makoto Kudo; Takeshi Kaneko; Yoshiaki Ishigatsubo
Abstract:Purpose. The predictive factor of response to antituberculous therapy has not been fully elucidated. Airway acidity has been thought to be a potential indicator of the bactericidal activity. Therefore, we hypothesized that monitoring airway acidity by measuring sputum pH could predict response to therapy. Methods. A total of 47 patients having newly diagnosed, smear-positive, active pulmonary tuberculosis were enrolled between October 2011 and March 2014. Sputum samples were serially analyzed before and after treatment. Eligible patients who initiated a standard 6-month treatment were monitored for the length of time to sputum smear and culture conversion. Results. There were 39 patients who completed a 2-month intensive phase of isoniazid, rifampicin, pyrazinamide, and ethambutol therapy followed by a 4-month continuation phase of isoniazid and rifampicin. Although factors including age, cavitation, sputum grade, and use of an acid-suppressant were associated with initial low sputum pH in univariate analysis, multivariate analysis revealed that only age ≥61 years was a statistically important factor predicting low pH value (p = 0.005). Further outcome analysis showed that initial low sputum pH before treatment was the only factor significantly associated with shorter length of time to both sputum smear and culture conversion (p = 0.034 and 0.019, respectively) independent of the effects of age, sputum bacterial load, extent of lung lesion, and cavitation. Thus, initial low sputum pH indicated favorable response to anti-tuberculosis therapy. Conclusions. Measuring sputum pH is an easy and inexpensive way of predicting response to standard combination therapy in patients with pulmonary tuberculosis.


An Autopsy Case Report of Malignant Pleural Mesothelioma with Deciduoid Features
INTERNAL MEDICINE,54(22):2915-2917 2015
Author:Ryota Ushio; Masaki Yamamoto; Yuji Shibata; Hiroshi Ishii; Keisuke Watanabe; Ryohei Takahashi; Takashi Sato; Makoto Kudo; Akio Miyake; Takeshi Kaneko; Yoshiaki Ishigatsubo
Abstract:Deciduoid mesothelioma is a rare variant of epithelioid mesothelioma. We experienced the case of a 73-year-old man with asbestos exposure who was diagnosed with malignant pleural mesothelioma with deciduoid features. He received chemotherapy containing six cycles of cisplatin and pemetrexed and survived for twenty-five months after the diagnosis. At autopsy, the final diagnosis was biphasic pleural mesothelioma. Cells with deciduoid features had mostly disappeared, and spindle cells markedly proliferated. To the best of our knowledge, this is the first autopsy case of malignant pleural mesothelioma with deciduoid features that exhibited a response to chemotherapy.


IKK alpha negatively regulates ASC-dependent inflammasome activation
NATURE COMMUNICATIONS,5:4977 2014(Sep.)
Author:Bradley N. Martin; Chenhui Wang; Jami Willette-Brown; Tomasz Herjan; Muhammet F. Gulen; Hao Zhou; Katarzyna Bulek; Luigi Franchi; Takashi Sato; Emad S. Alnemri; Goutham Narla; Xiao-Ping Zhong; James Thomas; Dennis Klinman; Katherine A. Fitzgerald; Michael Karin; Gabriel Nunez; George Dubyak; Yinling Hu; Xiaoxia Li
Abstract:The inflammasomes are multiprotein complexes that activate caspase-1 in response to infections and stress, resulting in the secretion of pro-inflammatory cytokines. Here we report that I kappa B kinase alpha (IKK alpha) is a critical negative regulator of apoptosis-associated specklike protein containing a C-terminal caspase-activation-andrecruitment ( CARD) domain ( ASC)-dependent inflammasomes. IKK alpha controls the inflammasome at the level of the adaptor ASC, which interacts with IKK alpha in the nucleus of resting macrophages in an IKK alpha kinase-dependent manner. Loss of IKK alpha kinase activity results in inflammasome hyperactivation. Mechanistically, the downstream nuclear effector IKK-related kinase ( IKKi) facilitates translocation of ASC from the nucleus to the perinuclear area during inflammasome activation. ASC remains under the control of IKK alpha in the perinuclear area following translocation of the ASC/IKK alpha complex. Signal 2 of NLRP3 activation leads to inhibition of IKK alpha kinase activity through the recruitment of PP2A, allowing ASC to participate in NLRP3 inflammasome assembly. Taken together, these findings reveal a IKKi-IKK alpha-ASC axis that serves as a common regulatory mechanism for ASC-dependent inflammasomes.


Generation of Dipeptidyl Peptidase-IV-Inhibiting Peptides from beta-Lactoglobulin Secreted by Lactococcus lactis
BIOMED RESEARCH INTERNATIONAL,2014:393598 2014
Author:Suguru Shigemori; Kazushi Oshiro; Pengfei Wang; Yoshinari Yamamoto; Yeqin Wang; Takashi Sato; Yutaka Uyeno; Takeshi Shimosato
Abstract:Previous studies showed that hydrolysates of beta-lactoglobulin (BLG) prepared using gastrointestinal proteases strongly inhibit dipeptidyl peptidase-IV (DPP-IV) activity in vitro. In this study, we developed a BLG-secreting Lactococcus lactis strain as a delivery vehicle and in situ expression system. Interestingly, trypsin-digested recombinant BLG from L. lactis inhibited DPP-IV activity, suggesting that BLG-secreting L. lactis may be useful in the treatment of type 2 diabetes mellitus.


Expression of the immunoreactive buckwheat major allergenic storage protein in Lactococcus lactis
Applied Microbiology and Biotechnology,97(8):3603-3611 2013(Apr.)
Author:Suguru Shigemori; Shinichi Yonekura; Takashi Sato; Hajime Otani; Takeshi Shimosato


Suppressive oligodeoxynucleotides synergistically enhance antiproliferative effects of anticancer drugs in A549 human lung cancer cells
International Journal of Oncology,42(2):429-436 2013(Feb.)
Author:Ryohei Takahashi; Takashi Sato; Dennis M. Klinman; Takeshi Shimosato; Takeshi Kaneko; Yoshiaki Ishigatsubo
Abstract:Immunosuppressive oligodeoxynucleotides (Sup ODNs) containing repetitive TTAGGG motifs reduce inflammation and, thus, may have an impact on inflammation-related tumor growth. In this study, we found a significant antiproliferative effect of Sup ODNs on the A549 non-small cell lung cancer (NSCLC) cell line compared to those treated with control ODNs (p< 0.05). Sup-ODN-mediated G1 phase cell cycle arrest was achieved via inhibition of Akt and extracellular signal-regulated kinase 1/2 phosphorylation and the p15INK4b and p27KIP1/retinoblastoma protein pathway. In addition, Sup ODNs induced apoptosis and enhanced apoptosis when combined with vinorelbine. In a setting similar to clinical use of multidrug chemotherapy for advanced NSCLC, these effects were investigated by using Sup ODNs in combination with conventional anticancer drugs. Sup ODNs had a significant synergistic effect with 5-fluorouracil, vinorelbine, gemcitabine, paclitaxel and irinotecan, with a mean combination index of 0.43-0.78 (< 1.0 indicates synergism) in the A549 NSCLC cell line. In conclusion, our results showed that Sup ODNs have an anticancer effect and increase the sensitivity of NSCLC cells to conventional anticancer drugs by modifying Akt and the extracellular signal-regulated kinase 1/2 pathway. Thus, Sup ODNs may serve as a novel therapeutic strategy for NSCLC patients.


Development and validation of a tuberculosis prognostic score for smear-positive in-patients in Japan
INTERNATIONAL JOURNAL OF TUBERCULOSIS AND LUNG DISEASE,17(1):54-60 2013(Jan.)
Author:N. Horita; N. Miyazawa; T. Yoshiyama; T. Sato; M. Yamamoto; K. Tomaru; M. Masuda; K. Tashiro; M. Sasaki; S. Morita; T. Kaneko; Y. Ishigatsubo
Abstract:BACKGROUND: No scoring system has ever been used to estimate the prognosis of individual tuberculosis (TB) patients. OBJECTIVE: To develop and validate a tuberculosis prognostic score. METHODS: This retrospective cohort study conducted in Japan comprised the development (n = 179; mean age 65.9 +/- 18.8 years) and validation (n = 244; mean age 64.3 +/- 20.1 years) of a tuberculosis prognostic score among patients with newly diagnosed smear-positive non-multidrug-resistant pulmonary tuberculosis without human immunodeficiency virus infection. The score (raw score) was defined by modifying a logistic regression formula using known risk factors as independent variables and in-patient death as a dependent variable. RESULTS: The raw score was calculated as follows: age (years) + (oxygen requirement, 10 points) -20 x albumin (g/dl) + (activity of daily living: independent, 0 point; semi-dependent, 5 points; totally dependent, 10 points). The raw scores were grouped into risk groups 1 (raw score < -30) to 5 (raw score >= 60) using 30-point intervals. Every increase in risk group was equivalent to a 7.3-fold increase in the odds ratio for in-hospital death (P < 0.001). The area under the receiver operating characteristics curve by risk group for in-patient death was 0.875 (P < 0.001). CONCLUSIONS: In this study we were able to develop and validate a tuberculosis prognostic score.


Class I/II hybrid inhibitory oligodeoxynucleotide exerts Th1 and Th2 double immunosuppression
FEBS OPEN BIO,3:41-45 2013
Author:Yusuke Ito; Suguru Shigemori; Takashi Sato; Tomoyuki Shimazu; Konomi Hatano; Hajime Otani; Haruki Kitazawa; Takeshi Shimosato
Abstract:We designed class I/II hybrid inhibitory oligodeoxynucleotides (iODNs), called iSG, and found that the sequence 5'-TTAGGG-3', which has a six-base loop head structure, and a 3'-oligo (dG)(3-5) tail sequence are important for potent immunosuppressive activity. Interestingly, splenocytes isolated from ovalbumin (OVA)-immunized mice and treated with iSG3 showed suppression of not only interleukin (IL)-6, IL-12p35, IL-12p40, and interferon (IFN) gamma mRNA expression, but also IL-4 and IL-13 mRNA expression. Thus, both Th2 and Th1 immune responses can be strongly suppressed by iODNs in splenocytes from allergen-immunized mice, suggesting usefulness in the treatment of diseases induced by over-active immune activation. (C) 2012 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.


Serum Heme Oxygenase-1 as a Marker of Lung Function Decline in Patients With Chronic Silicosis
JOURNAL OF OCCUPATIONAL AND ENVIRONMENTAL MEDICINE,54(12):1461-1466 2012(Dec.)
Author:Takashi Sato; Yoshiaki Saito; Satoshi Inoue; Takeshi Shimosato; Shigeto Takagi; Takeshi Kaneko; Yoshiaki Ishigatsubo
Abstract:Objective: To identify predictive factors of excess decline in forced expiratory volume in one second (FEV1) in patients with chronic silicosis. Methods: Forty-six male patients enrolled in 2004 were screened and received pulmonary function tests. Results: Among the 33 included patients, 12 were categorized as rapid decliners (reduction in FEV1 > 60 mL/yr). The mean level of serum heme oxygenase-1 (HO-1), a marker of oxidative stress, was significantly lower in rapid decliners than in normal decliners (P = 0.002). Logistic regression analysis revealed that serum HO-1 was a factor affecting clinically important decline in FEV1 (odds ratio = 0.52; 95% confidence interval, 0.31 to 0.88) independent of the effects of age, height, weight, smoking, exposure status, and C-reactive protein. Conclusions: Serum HO-1 may be a predictor of lung function decline in silicosis patients.


Expression of a Biologically Active GFP-alpha(S1)-Casein Fusion Protein in Lactococcus lactis
CURRENT MICROBIOLOGY,64(6):569-575 2012(Jun.)
Author:Suguru Shigemori; Shinichi Yonekura; Takashi Sato; Maya Nakanishi; Hajime Otani; Takeshi Shimosato
Abstract:In this study, we successfully developed a recombinant strain of Lactococcus lactis NZ9000 (NZ9000) that produced green fluorescent protein fused to alpha(S1)-casein (GFP-alpha(S1)Cas). A modified lactic acid bacterial vector (pNZ8148#2) was constructed by inserting genes for GFP and alpha(S1)-casein, a major cow's milk allergen, and the resulting vector, pNZ8148#2-GFP-alpha(S1)Cas, was applied to the expression of recombinant GFP-alpha(S1)Cas protein (rGFP-alpha(S1)Cas) in NZ9000. After inducing expression with nisin, the production of rGFP-alpha(S1)Cas was confirmed by confocal laser microscopic analysis, and the expression conditions were optimized based on fluorescent analysis and western blotting results. Moreover, the in vitro treatment of splenocytes isolated from alpha-casein (a parts per thousand yen70 % alpha(S)-casein)-immunized mice with rGFP-alpha(S1)Cas resulted in increased IL-13 mRNA expression. The observed allergic activity is indicative of the Th2-cell mediated immune response and is similar to the effects induced by exposure to alpha-casein. Our results suggest that the expression of rGFP-alpha(S1)Cas in NZ9000 may facilitate in vivo applications of this system aimed at improving the specificity of immunological responses to specific milk allergen.


Synchronous lung and gastric cancers successfully treated with carboplatin and pemetrexed: A case report
Journal of Medical Case Reports,6:266 2012
Author:Takashi Sato; Koji Tomaru; Tomoko Koide; Makoto Masuda; Masaki Yamamoto; Naoki Miyazawa; Yoshiaki Inayama; Takeshi Kaneko; Yoshiaki Ishigatsubo
Abstract:Introduction: Lung and gastric cancers are the first and second leading causes of death from cancer worldwide, and are especially prevalent in Eastern Asia. Relatively few reports are available in relation to the treatment and outcome of synchronous lung and gastric cancers, although there are increasing numbers of patients with these cancers. Efforts to develop more effective drugs for the treatment of synchronous cancers, without serious adverse effects, have been intensifying. Pemetrexed, a multi-targeted antifolate enzyme inhibitor, was approved by the United States Food and Drug Administration as a first-line chemotherapy for advanced non-squamous non-small cell lung cancer in 2007. Although clinical activity against several tumor types of adenocarcinoma, including gastric cancer, has been demonstrated, the efficacy of pemetrexed for gastric cancer remains to be fully evaluated. Case presentation: We report a case involving a 62-year-old Japanese woman with synchronous locally-advanced poorly-differentiated lung adenocarcinoma and poorly-differentiated gastric adenocarcinoma, containing signet-ring cells distinguished by immunohistochemical profiles. She had been treated with carboplatin and pemetrexed as a first-line chemotherapy for lung cancer, and had achieved partial responses for both lung and gastric cancers. These responses led to a favorable 12-month progression-free survival after the initiation of chemotherapy, and the patient is still alive more than 33 months after diagnosis. Conclusions: This case suggests a new chemotherapeutic regimen for patients with synchronous multiple primary cancers that have an adenocarcinoma background. © 2012 Sato et al. licensee BioMed Central Ltd.


The acceleration of wound healing in primates by the local administration of immunostimulatory CpG oligonucleotides
BIOMATERIALS,32(18):4238-4242 2011(Jun.)
Author:Masaki Yamamoto; Takashi Sato; Joel Beren; Daniela Verthelyi; Dennis M. Klinman
Abstract:The process of wound healing involves complex interactions between circulating immune cells and local epithelial and endothelial cells. Studies in murine models indicate that cells of the innate immune system activated via their Toll-like receptors (TLR) can accelerate wound healing. This work examines whether immunostimulatory CpG oligodeoxynucleotides (ODN) designed to trigger human immune cells via TLR9 can promote the healing of excisional skin biopsies in rhesus macaques. Results indicate that 'K' type CpG ODN significantly accelerate wound closure in non-human primates (p < 0.05). Contributing to this outcome was a CpG-dependent increase in both the production of basic fibroblast growth factor and in keratinocyte migration. Of interest, IL-1 alpha and TGF alpha normally present at sites of skin injury facilitated these effects. Current findings support the conclusion that the local administration of CpG ODN may provide an effective strategy for accelerating wound healing in humans. Published by Elsevier Ltd.


Serum Heme Oxygenase-1 As Marker Of Lung Function Decline In Patients With Chronic Silicosis
AMERICAN JOURNAL OF RESPIRATORY AND CRITICAL CARE MEDICINE,183(12):1461 2011
Author:T. Sato; N. Miyazawa; Y. Saito; R. Takahashi; T. Tsukahara; M. Hayashi; M. Yamamoto; K. Tomaru; M. Sasaki; T. Kaneko; Y. Ishigatsubo


Heterogeneity of the efficacy of the 23-valent pneumococcal polysaccharide vaccine caused by various underlying conditions of chronic pulmonary disease in older patients: prospective cohort study
BMJ OPEN,1(1):e000105 2011
Author:Satoshi Inoue; Yuji Watanuki; Tetsuji Kaneko; Takashi Sato; Naoki Miyazawa; Takeshi Kaneko; Yoshiaki Ishigatsubo; Satoshi Morita; Yutaka Natsumeda; Shunsaku Mizushima
Abstract:Objective: To determine the ideal conditions for use of the 23-valent pneumococcal polysaccharide vaccine (PPV23) in older outpatients with chronic pulmonary diseases. Design: Prospective cohort study. Participants: 1378 outpatients with chronic pulmonary diseases >= 60 years of age. Intervention: Participants were educated about PPV23, and those who responded affirmatively were vaccinated between August and November 2002. The participants who chose no intervention served as controls. The prevaccine period was defined as August 2001 to August 2002. Participants were followed for 2 years from December 2002 or until death. Main outcome measures: Events of interest included the first episode of bacterial (including pneumococcal) pulmonary infection (primary endpoint) and death of any cause (secondary endpoint). Results: Frequent episodes of pulmonary infection during the prevaccine period significantly decreased event-free survival during the 2-year observation period (p<0.001). Chronic respiratory failure was associated with a decreased event-free survival only when the pulmonary infection episode did not occur in the prevaccine period (p<0.001). No significant differences in event-free survival were observed between the vaccinated and unvaccinated group during analysis of the entire cohort. In the Cox proportional hazards regression model, event-free survival decreased significantly when pulmonary infection occurred in the prevaccine period. In the subgroup analysis, the first episode of bacterial pulmonary infection (but not death of any cause) was reduced significantly by PPV23 only in patients with chronic respiratory failure who had no episodes of pulmonary infection during the prevaccine period (p=0.019). Conclusion: The efficacy of PPV23 against pulmonary infection and death of any cause might be unachievable if pulmonary infection occurs during the prevaccine period. PPV23 needs to be given to older patients with chronic pulmonary disease at an earlier time in which infectious complications in the lung have not yet occurred.


Accelerated wound healing mediated by activation of Toll-like receptor 9
WOUND REPAIR AND REGENERATION,18(6):586-593 2010(Nov.)
Author:Takashi Sato; Masaki Yamamoto; Takeshi Shimosato; Dennis M. Klinman
Abstract:Wound healing is mediated through complex interactions between circulating immune cells and local epithelial and endothelial cells. Elements of the innate immune system are triggered when Toll-like receptors (TLR) are stimulated by their cognate ligands, and previous studies suggest that such interactions can accelerate wound healing. This work examines the effect of treating excisional skin biopsies with immunostimulatory CpG oligodeoxynucleotides (ODN) that trigger via TLR9. Results indicate that CpG (but not control) ODN accelerate wound closure and reduce the total wound area exposed over time by > 40% (p < 0.01). TLR9 knockout mice, a strain unresponsive to the immunomodulatory effects of CpG stimulation, are unresponsive to ODN treatment and exhibit a general delay in healing when compared with wild-type mice. CpG ODN administration promoted the influx of macrophages to the wound site and increased the production of vascular endothelial growth factor, expediting neovascularization of the wound bed (p < 0.01 for both parameters). Stimulation via TLR9 thus represents a novel strategy to accelerate wound healing.


CpG oligodeoxynucleotides induce strong up-regulation of interleukin 33 via Toll-like receptor 9
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS,394(1):81-86 2010(Mar.)
Author:Takeshi Shimosato; Megumi Fujimoto; Masanori Tohno; Takashi Sato; Mariko Tateo; Hajime Otani; Haruki Kitazawa
Abstract:We previously reported the strong immunostimulatory effects of a CpG oligodeoxynucleotide (ODN), designated MsST, from the lacZ gene of Streptococcus (S.) thermophilus ATCC19258. Here we show that 24 h of stimulation with MsST in mouse splenocytes and peritoneal macrophages strongly induces expression of interleukin (IL)-33, a cytokine in the IL-1 superfamily. Other IL-1 superfamily members, including IL-1 alpha, IL-1 beta and IL-18, are down-regulated after 24 h of stimulation of MsST. We also found that MsST-induced IL-33 mRNA expression is inhibited by the suppressive ODN A151, which can inhibit Toll-like receptor 9 (TLR9)-mediated responses. This is the first report to show that IL-33 can be induced by CpG ODNs. The strong induction of IL-33 by MsST suggests that it may be a potential therapeutic ODN for the treatment of inflammatory disease. The presence of a strong CpG ODN in S. thermophilus also suggests that the bacterium may be a good candidate as a starter culture for the development of new physiologically functional foods. (C) 2010 Elsevier Inc. All rights reserved.


High frequency of beta-lactamase-negative, ampicillin-resistant strains of Haemophilus influenzae in patients with chronic bronchitis in Japan
JOURNAL OF INFECTION AND CHEMOTHERAPY,16(1):72-75 2010(Feb.)
Author:Satoshi Inoue; Yuji Watanuki; Naoki Miyazawa; Makoto Kudo; Takashi Sato; Nobuaki Kobayashi; Kei Mishina; Masahiro Sasaki; Takeshi Kaneko; Yoshiaki Ishigatsubo
Abstract:In Japan, the increasing incidence of beta-lactum-resistant Haemophilus influenzae infections is of growing concern. We retrospectively studied whether the prevalence of beta-lactamase-negative ampicillin-resistant strains of H. influenzae was influenced by chronic lung diseases. H. influenzae isolates, obtained from patients who were diagnosed with acute or chronic bronchitis, or acute exacerbation of chronic bronchitis in 2005, were studied. In addition to susceptibility testing, polymerase chain reaction (PCR) was performed for the detection of TEM-1 beta-lactamase, and Asn526-Lys and Ser385-Thr amino acid substitutions in the ftsI gene encoding penicillin-binding protein-3 (PBP-3). The minimum inhibitory concentration values of beta-lactams were found to be increased in isolates from patients with chronic bronchitis who had been repeatedly administered antibiotics. Genetic analysis using PCR suggested that this might be associated with a high frequency of beta-lactamase-negative strains with mutations in PBP-3. The presence of beta-lactum-resistant strains needs to be considered for patients with chronic bronchitis in whom H. influenzae is isolated as a causative pathogen.


Esophagomediastinal and esophagobronchial fistulas associated with invasive aspergillosis.
Endoscopy,42 Suppl 2:E48-9 2010
Author:Kato S; Inoue S; Inamori M; Miyazawa N; Sato T; Kobayashi N; Mishina K; Sasaki M; Ishigatsubo Y; Nakajima A


[The preventive effect of 23-valent pneumococcal polysaccharide vaccine against drug-resistant Streptococcus pneumonia in patients with chronic respiratory disease].
Nihon Kokyuki Gakkai zasshi = the journal of the Japanese Respiratory Society,47(12):1063-1069 2009(Dec.)
Author:Kobayashi N; Watanuki Y; Miyazawa N; Kudo M; Inoue S; Sato T; Mishina K; Takahashi H; Kaneko T; Ishigatsubo Y


CpG oligonucleotides as adjuvants for vaccines targeting infectious diseases
ADVANCED DRUG DELIVERY REVIEWS,61(3):248-255 2009(Mar.)
Author:Dennis M. Klinman; Sven Klaschik; Takashi Sato; Debbie Tross
Abstract:Synthetic oligodeoxynucleotides (ODN) containing unmethylated CpG motifs act as immune adjuvants, accelerating and boosting antigen-specific immune responses. CpG motifs promote the induction of Th1 and pro-inflammatory cytokines and support the maturation/activation of professional antigen presenting cells (particularly plasmacytoid dendritic cells). These effects are optimized by maintaining close physical contact between the CpG ODN and the immunogen. Co-administering CpG ODN with a variety of vaccines has improved the resultant humoral and/or cellular immune responses, culminating in enhanced protective immunity in rodent and primate challenge models. Ongoing clinical studies indicate that CpG ODN are safe and well-tolerated when administered as adjuvants to humans, and that they can support increased vaccine-specific immune responses. Published by Elsevier B.V.


Therapeutic Applications and Mechanisms Underlying the Activity of Immunosuppressive Oligonucleotides
OLIGONUCLEOTIDE THERAPEUTICS,1175:80-88 2009
Author:Dennis M. Klinman; Debbie Tross; Sven Klaschik; Hidekazu Shirota; Takeshi Sato
Abstract:Synthetic oligodeoxynucleotides (ODN) capable of "neutralizing" or "inhibiting" immune responses have been described. This review will focus on the properties of phosphorothioate ODN that mimic the immunosuppressive activity of the repetitive TTAGGG motifs present in mammalian telomeres. These TTAGGG multimers block the production of pro-inflammatory and T helper type 1 cytokines elicited when immune cells are activated by a wide variety of Toll-like receptor ligands, polyclonal activators, and antigens. Several mechanisms contribute to the suppressive activity of such ODN. Ongoing microarray studies indicate that suppressive ODN interfere with the phosphorylation of signal transducer and activator of transcription I (STATI) and STAT4, thereby blocking the inflammation mediated by STAT-associated signaling cascades. In animal models, suppressive ODN can be used to prevent or treat diseases characterized by persistent immune activation, including collagen-induced arthritis, inflammatory arthritis, systemic lupus erythematosus, silicosis, and toxic shock. These findings suggest that TTAGGG multimers may find broad use in the treatment of diseases characterized by over-exuberant/persistent immune activation.


Identification of a potent immunostimulatory oligodeoxynucleotide from Streptococcus thermophilus lacZ
ANIMAL SCIENCE JOURNAL,80(5):597-604 2009
Author:Takeshi Shimosato; Masanori Tohno; Takashi Sato; Junko Nishimura; Yasushi Kawai; Tadao Saito; Haruki Kitazawa
Abstract:Immunostimulatory sequences of oligodeoxynucleotides (ODNs), such as CpG ODNs, are potent stimulators of innate immunity. Here, we identified a strong immunostimulatory CpG ODN, which we named MsST, from the lac Z gene of Streptococcus (S.) thermophilus ATCC19258, and we evaluated its immune functions. In in vitro studies, MsST had a similar ability as the murine prototype CpG ODN 1555 to induce inflammatory cytokine production and cell proliferation. In mouse splenocytes, MsST increased the number of CD80+CD11c+and CD86+CD11c+ dendritic cells and CD4+CD25+ regulatory T cells. We also analyzed the effects of MsST on the expression of regulatory cytokines by real-time quantitative PCR. MsST was more potent at inducing interleukin-10 expression than the ODN control 1612, indicating that MsST can augment the regulatory T cell response via Toll-like receptor 9, which plays an important role in suppressing T helper type 2 responses. These results suggest that S. thermophilus, whose genes include a strong Immunostimulatory sequence-ODN, is a good candidate for a starter culture to develop new physiologically functional foods and feeds.


Synthetic oligonucleotides as modulators of inflammation
JOURNAL OF LEUKOCYTE BIOLOGY,84(4):958-964 2008(Oct.)
Author:Dennis Klinman; Hidekazu Shirota; Debra Tross; Takashi Sato; Sven Klaschik
Abstract:Synthetic oligodeoxynucleotides (ODN) containing unmethylated CpG motifs mimic the immunostimulatory activity of bacterial DNA. CpG ODN directly stimulate human B cells and plasmacytoid dendritic cells, promote the production of Th1 and proinflammatory cytokines, and trigger the maturation/activation of professional APC. CpG ODN are finding use in the treatment of cancer, allergy, and infection. In contrast, ODN containing multiple TTAGGG motifs mimic the immunosuppressive activity of self-DNA, down-regulating the production of proinflammatory and Th1 cytokines. Preclinical studies suggest that "suppressive" ODN may slow or prevent diseases characterized by pathologic immune stimulation, including autoimmunity and septic shock. Extensive studies in animal models suggest that the therapeutic value of CpG and TTAGGG ODN may be optimized by early administration.


Suppressive oligodeoxynucleotides inhibit silica-induced pulmonary inflammation
JOURNAL OF IMMUNOLOGY,180(11):7648-7654 2008(Jun.)
Author:Takashi Sato; Takeshi Shimosato; W. Gregory Alvord; Dennis M. Klinman
Abstract:Inhalation of silica-containing dust particles induces silicosis, an inflammatory disease of the lungs characterized by the infiltration of macrophages and neutrophils into the lungs and the production of proinflammatory cytokines, chemokines, and reactive oxygen species (ROS). Synthetic oligodeoxynucleotides (ODN) expressing "immunosuppressive motifs" were recently shown to block pathologic inflammatory reactions in murine models of autoinumme disease. Based on those findings, the potential of suppressive ODN to prevent acute murine silicosis was examined. In vitro studies indicate that suppressive ODN blunt silica-induced macrophage toxicity. This effect was associated with a reduction in ROS production and p47phox expression (a subunit of NADPH oxidase key to ROS generation). In vivo studies show that pretreatment with suppressive (but not control) ODN reduces silicadependent pulmonary inflammation, as manifest by fewer infiltrating cells, less cytokine/chemokine production, and lower levels of ROS (p < 0.01 for all parameters). Treatment with suppressive ODN also reduced disease severity and improved the survival (p < 0.05) of mice exposed to silica.


Heme oxygenase-1, a potential biomarker of chronic silicosis, attenuates silica-induced lung injury
AMERICAN JOURNAL OF RESPIRATORY AND CRITICAL CARE MEDICINE,174(8):906-914 2006(Oct.)
Author:Takashi Sato; Mitsuhiro Takeno; Koichi Honma; Hicleyuki Yamauchi; Yoshiaki Saito; Takao Sasaki; Hiroshi Morikubo; Yoji Nagashima; Shigeto Takagi; Kouichi Yamanaka; Takeshi Kaneko; Yoshiaki Ishigatsubo
Abstract:Rationale: Heme oxygenase-1 (HO-1), a rate-limiting enzyme in heme catabolism, has antioxidative, antiapoptotic, and antiinflammatory activities. We examined whether HO-1 might be involved in silicosis. Objectives: To investigate whether HO-1 can reduce silicosis in mice and humans. Methods and measurements: Silicosis was studied using a murine model, and in 46 male patients. Serum HO-1 and 8-hydroxydeoxyguanosine (a marker of oxidative stress) were measured by enzyme-linked immunosorbent assay. Levels of HO-1 were measured by immunohistochemistry and immunoblotting. Main results: Serum HO-1 levels were significantly elevated in patients with silicosis compared with age-matched control subjects or patients with chronic obstructive pulmonary disease. Serum HO-1 levels also correlated inversely with serum 8-hydroxydeoxyguanosine levels and positively with vital capacity and forced expiratory volume in one second in patients with silicosis. HO-1 was present in the lungs of humans and mice with silicosis, especially at sites of silica particle deposition. In mice, silica exposure was associated with acute leukocyte infiltration, leading to development of silicotic lung lesions. The inflammation was suppressed by treatment with hemin, an inducer of HO-1, and enhanced by zinc protoporphyrin, an inhibitor of HO-1. Conclusions: Pulmonary HO-1 expression is increased in silicosis. HO-1 suppresses reactive oxygen species activity, and subsequent pathologic changes, thereby attenuating disease progression.


Heme oxygenase-1 inhibits cigarette smoke-induced increase in the tracheal mucosal permeability in guinea pigs in vivo
INFLAMMATION RESEARCH,54(5):229-234 2005(May)
Author:A Tagawa; T Kaneko; T Shinohara; A Ueda; T Sato; Y Ishigatsubo
Abstract:Objective: To examine whether heme oxygenase (HO)-1 inhibits cigarette smoke (CS)-induced increase in the airway mucosal permeability. Methods: Mucosal permeability was quantified by monitoring the rate of appearance in the circulation of horseradish peroxidase that had been instilled into the isolated tracheal segment in guinea pigs in vivo, after exposure to CS or room air. Results: Exposure to 10 puffs of C S did not increase the tracheal mucosal permeability but did increase the permeability after pretreatment with zinc protoporphyrin, a competitive inhibitor of HO-1. Moreover, pretreatment with hemin, a potent inducer of HO-1, inhibited the increase in the permeability of the tracheal mucosa induced by 20 puffs of CS exposure. Conclusion: It is concluded that HO-1 has an important role in suppressing the increase in the mucosal permeability induced by CS in guinea pig trachea.


Cigarette smoke increases mucosal permeability in guinea pig trachea via tachykinin NK2 receptor activation
EUROPEAN JOURNAL OF PHARMACOLOGY,507(1-3):223-228 2005(Jan.)
Author:A Tagawa; T Kaneko; H Nishiyama; T Shinohara; T Sato; P Geppetti; Y Ishigatsubo
Abstract:We investigated whether exposure to cigarette smoke increases the mucosal permeability in guinea pig trachea and if this effect could be mediated by tachykinin NK2 receptor activation. Guinea pigs were exposed to either three different doses of cigarette smoke or room air. Mucosal permeability was measured by monitoring the rate of appearance in the circulation of horseradish peroxidase (HRP) that had been instilled into the isolated tracheal segment. Exposure to 20 and 30 puffs but not 10 puffs of cigarette smoke increased the tracheal mucosal permeability. Pretreatment with the tachykinin NK2 receptor antagonist SR48,968 [(S)-N-methyl-N[4-(4-acetylamino-4-phenylpiperidino)-2-(3,4-dichlorophenyl)butyl]benzamide] completely inhibited the increase in the permeability of the tracheal mucosa induced by exposure to cigarette smoke, whereas the tachykinin NK1 receptor antagonist SSR240,600 [(R)-2-(1-{2-[4-{2-[3,5-bis(trifluoromethyl)phenyl]acetyl}-2-(3,4-dichlorophenyl)-2-morpholinyl]ethyl}-4-piperidinyl)-2-methylpropanamide] and the tachykinin NK3 receptor antagonist SR142,801 [(S)-(N)-(1-[3-(1-benzoyl-3(3,4-dichlorophenyl)piperidine-3-yl)propyl]-4-phenylpiperidin-4-yl)-N-methyl-acetamide] had no effect. It is concluded that endogenous tachykinins via NK2 receptor activation mediate the increase in the permeability of the tracheal mucosa induced by exposure to cigarette smoke in guinea pigs. (C) 2004 Elsevier B.V. All rights reserved.


Presentations
Intratracheal therapy using oligodeoxynucleotides and/or genetically modified lactic acid bacteria-derived biological molecules
The Korean Society of Toxicology/The Korean Environmental Mutagen Society 2022 KSOT/KEMS Spring International Symposium 2022(May 13)
Presenter:Takashi Sato


Innovative therapy for lung cancer ~Intratracheal delivery of nanomicroparticles-incorporated immune checkpoint-blocking single-chain variable fragment~
NIH/NCI Symposium on Innate Immunity & Modulation of the Host Immune Response in Honor of the Career of Dr. Dennis Klinman 2019(Jun. 27)
Presenter:SATO Takashi


吸入治療イノベーション~吸入薬で肺癌の制御を目指す研究開発の取り組み~
横浜市立大学先端医科学研究センター 第54回市民講座 2018(Oct. 30)
Presenter:佐藤 隆


Intratracheal delivery of immunostimulatory oligonucleotides using biodegradable polyketal nanoparticles: effect of long-term immune responses on murine lung cancer
THE 14TH US-JAPAN SYMPOSIUM ON DRUG DELIVERY SYSTEMS 2017(Dec. 16)
Presenter:Takashi Sato


Intratracheal delivery of immunostimulatory oligonucleotides using biodegradable polyketal nanoparticles: effect on murine lung cancer
THE 13TH US-JAPAN SYMPOSIUM ON DRUG DELIVERY SYSTEMS 2015(Dec. 18)
Presenter:Takashi Sato


MISC
Intranasal Administration of Genetically Modified Lactococcus Lactis Producing PD-L1 scFv Improves Survival in Lung Cancer Model Mice
C70. COPD (AND LUNG CANCER) CLINICAL AND TRANSLATIONAL STUDIES,207:A5754 2023(May 01)
Author:N. Nomura; F. Namai; T. Sato; T. Shimosato


Effect of Local Intratracheal Therapy Using Particleized Nintedanib in a Murine Model of Pulmonary Fibrosis
D29. TRANSLATIONAL RESEARCH IN IPF,207:A6476 2023(May 01)
Author:K. Suzuki; F. Namai; N. Nomura; Y. Sato; A. Kotsugai; T. Shimosato; T. Sato


A Case of Adenocarcinoma of the Lung With EGFR T790 Mutation Presenting With Chylous Ascites.
Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer,13(8):1227-1228 2018(Aug.)
Author:Kentaro Nakashima; Yu Hara; Takashi Sato; Masaharu Shinkai; Takeshi Kaneko


Serum Cxcl16 As A Potential Prognostic Marker In Advanced Non Small Cell Lung Cancer
AMERICAN JOURNAL OF RESPIRATORY AND CRITICAL CARE MEDICINE,195 2017
Author:Y. Shibata; T. Sato; M. Ikeda; H. Sasaki; A. Nagashima; R. Ushio; K. Nakashima; H. Watanabe; K. Nagai; K. Tashiro; H. Nagakura; N. Horita; Y. Hara; N. Kobayashi; M. Yamamoto; M. Kudo; M. Shinkai; T. Kaneko


Toll-Like Receptor 9, A Key To Attenuate Disease Progression In A Murine Model Of COPD
AMERICAN JOURNAL OF RESPIRATORY AND CRITICAL CARE MEDICINE,195 2017
Author:T. Sato; K. Nakashima; Y. Shibata; H. Sasaki; M. Ikeda; A. Nagashima; H. Watanabe; K. Nagai; H. Nagakura; N. Horita; N. Kobayashi; M. Yamamoto; M. Kudo; M. Shinkai; T. Kaneko


Heme Oxygenase-1 Regulates Ros-Erk1/2 Signaling In Silica-Induced Lung Injury
AMERICAN JOURNAL OF RESPIRATORY AND CRITICAL CARE MEDICINE,193 2016
Author:K. Nakashima; T. Sato; Y. Shibata; R. Ushio; M. Ikeda; H. Watanabe; K. Nagai; K. Tashiro; H. Nagakura; N. Horita; M. Yamamoto; M. Shinkai; M. Kudo; T. Kaneko


Toll-Like Receptor 9, A Key To Attenuation Of Disease Progression In A Mouse Model Of Chronic Obstructive Pulmonary Disease
AMERICAN JOURNAL OF RESPIRATORY AND CRITICAL CARE MEDICINE,193 2016
Author:T. Sato; K. Nakashima; Y. Shibata; R. Ushio; M. Ikeda; H. Watanabe; K. Nagai; K. Tashiro; H. Nagakura; N. Horita; M. Yamamoto; M. Shinkai; M. Kudo; D. Klinman; T. Kaneko


Effect Of Intratracheal Immunostimulatory-Oligonucleotides With Biodegradable Polyketal Nanoparticles On Murine Lung Cancer
AMERICAN JOURNAL OF RESPIRATORY AND CRITICAL CARE MEDICINE,191 2015
Author:T. Sato; Y. Shibata; M. Yamamoto; M. Kudo; T. Kaneko; Y. Ishigatsubo; D. Klinman


Serum Cxcl16, A Potential Prognostic Marker In Advanced Non Small Cell Lung Cancer: A Preliminary Report
AMERICAN JOURNAL OF RESPIRATORY AND CRITICAL CARE MEDICINE,191 2015
Author:Y. Shibata; T. Sato; M. Yamamoto; M. Kudo; T. Kaneko; Y. Ishigatsubo


CPG Oligodeoxynucleotides Promote Wound Healing In Airway Epithelial Cells
AMERICAN JOURNAL OF RESPIRATORY AND CRITICAL CARE MEDICINE,183 2011
Author:T. Sato; N. Miyazawa; D. M. Klinman; T. Shimosato; R. Takahashi; T. Tsukahara; M. Hayashi; M. Yamamoto; K. Tomaru; M. Sasaki; T. Kaneko; Y. Ishigatsubo


Suppressive Oligodeoxynucleotide Synergistically Enhances The Antiproliferative Effect Of Anti-Cancer Drug In Human Lung Cancer Cells
AMERICAN JOURNAL OF RESPIRATORY AND CRITICAL CARE MEDICINE,183 2011
Author:R. Takahashi; T. Sato; N. Miyazawa; D. M. Klinman; T. Shimosato; M. Hayashi; M. Yamamoto; K. Tomaru; T. Tsukahara; M. Sasaki; T. Kaneko; Y. Ishigatsubo


Accelerated wound healing in mice by local administration of sustained-release CpG oligodeoxynucleotide
JOURNAL OF IMMUNOLOGY,184 2010(Apr.)
Author:Takashi Sato; Takeshi Shimosato; Masaki Yamamoto; Nobuaki Kobayashi; Naoki Miyazawa; Yoshiaki Ishigatsubo; Dennis Klinman


CpG oligodeoxynucleotide incorporated basement membrane extract facilitate wound repair in mice
JOURNAL OF IMMUNOLOGY,182 2009(Apr.)
Author:Takashi Sato; Takeshi Shimosato; Dennis M. Klinman

Patents
Patents
Polyketal particles including a CpG oligodeoxynucleotide for the treatment of lung cancer
Method of treating pneumoconiosis with oligodeoxynucleotides
Use of CpG oligodeoxynucleotides to induce epithelial cell growth
Research Grants
Grants‐in‐aid for Scientific Research(Research Representative)
2019 - 2022 , 免疫チェックポイント阻害乳酸菌による肺癌発癌予防・進展予防効果の検証 , 国際共同研究加速基金(国際共同研究強化(B))
2018 - 2019 , 吸入療法イノベーションの実現:高分子・抗体医薬の経気道投与による肺癌治療戦略 , 挑戦的研究(萌芽)
2015 - 2017 , ナノ粒子化免疫調節剤の呼吸器疾患への治療応用の研究 , 基盤C
2011 - 2012 , 微生物由来 DNA を利用した吸入型炎症性肺疾患治療薬の開発 , 若手B
2009 - 2010 , 免疫調節性DNAとナノ・マイクロ粒子キャリアーを用いた吸入型DNA治療薬の開発 , 若手B

Grants‐in‐aid for Scientific Research(Researcher)
2017 - 2020 , 乳酸菌オリゴDNAを有効成分とする経口用ナノカプセルを用いた免疫調節機構の解明 , 基盤B

Other Research Grants
2017 - 2019 , 日本医療研究開発機構(AMED)・創薬基盤推進研究事業・Fine Droplet Dryingテクノロジーによる経気道治療用ナノマイクロ粒子包埋医薬品の創出を目指した研究
2017 - 2019 , ライフイノベーションプラットフォーム横浜推進プロジェクト・圧電素子を用いた経気道投与用パーソナル吸入デバイスの開発研究