The analysis of mechanisms of how leptospires invade the human body and cause the severe condition and the development of prophylaxis against leptospirosis

• Project/Area Number: 25460536
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Bacteriology (including mycology)
• Research Institution: University of Occupational and Environmental Health, Japan (2015); Kyushu University (2013-2014)
• Principal Investigator: SAITO Mitsumasa 産業医科大学, 医学部, 教授 (00315087)
• Co-Investigator(Kenkyū-buntansha): IIDA Ken-ichiro 九州大学, 大学院医学研究院, 助教 (00346777)
• Research Collaborator: Villanueva Sharon Y. A. M. ; ASO Tatsuma ; MIYAHARA Satoshi ; OZURU Ryo ; KANEMARU Takaaki ; SEGAWA Takaya ; YOSHIMURA Michinobu ; IKEJIRI Mami ; ARAMAKI Natsumi ; HIDAKA Yusuke ; Chakraborty Antara ; Widiyanti Dian ; Muslich Lisa Tenriesa ; Amran Muhammad Yunus ; Gloriani Nina G. ; KOBAYASHI Yoshie ; FUKUI Takashi ; MASUZAWA Toshiyuki ; YANAGIHARA Yasutake ; YOSHIDA Shin-ichi
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2015: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2014: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2013: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
• Keywords: レプトスピラ / ワイル病 / 黄疸出血性レプトスピラ症 / レプトスピラ感染症 / Luciferase / Leptospira interrogans / in vivo imaging system / 脂肪組織 / 肝細胞 / 胆汁排泄障害 / 黄疸 / 細胞間接着 / 肺出血 / 腎不全 / 臓器内菌数 / 出血

Outline of Final Research Achievements

We clarified the mechanisms of how leptospires invade the human body and cause the severe condition. Leptospires invade human and animal skin through the site without stratum corneum. They migrate through keratinocyte layers and reach the hypodermis. In early phase of infection, they colonize the blood vessels in adipose tissue and start to grow. After sufficient increase on the adipose tissue, leptospires move via the bloodstream and reach liver. Then, they infiltrate the Disse’s space, migrate between hepatocytes, detach the intercellular junctions and disrupt the bile canaliculi. Destruction of bile canaliculi coincide with the elevation of conjugated bilirubin levels in serum. It is expected that the analogues of target molecules for vascular colonization and hepatocytes invasion will prevent the progression to severe leptospirosis.

Research Products

• [Journal Article] The usefulness of semi-solid medium in the isolation of highly virulent Leptospira strains from wild rats in an urban area of Fukuoka, Japan. (2015)
  • Author(s): Saito M, Villanueva SY, Masuzawa T, Haraguchi Y, Ita S. Miyahara S, Ozuru R, Yamaguchi T, Yoshimura M, Ikejiri M, Aramaki N, Amran MY, Muslich LT, Iida K, Yanagihara Y, Gloriani NG, Yoshida S
  • Journal Title: Micribiol Immunol Volume: 59 Issue: 6 Pages: 322-30
  • DOI: 10.1111/1348-0421.12260
  • NAID: 40020510062
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Characterization of Leptospira infection in suckling and weaning rat pups. (2015)
  • Author(s): Muslich LT, Villanueva SY, Amran MY, Segawa T, Saito M, Yoshida S
  • Journal Title: Comp Immunol Microbiol Infect Dis Volume: 38 Pages: 47-55
  • DOI: 10.1016/j.cimid.2014.11.001
  • Peer Reviewed
• [Journal Article] Destruction of the hepatocyte junction by intercellular invasion of Leptospira causes jaundice in a hamster model of Weil’s disease. (2014)
  • Author(s): Miyahara S, Saito M, Kanemaru T, Villanueva SY, Gloriani NG, Yoshida S
  • Journal Title: Int J Exp Pathol Volume: 95 Issue: 4 Pages: 271-281
  • DOI: 10.1111/iep.12085
  • Peer Reviewed
• [Journal Article] Natural Defense by Saliva and Mucosa against Oral Infection by Leptospira. (2014)
  • Author(s): Asoh T, Saito M, Villanueva SY, Kanemaru T, Gloriani NG, Yoshida S
  • Journal Title: Can J Microbiol Volume: 60 Issue: 6 Pages: 383-389
  • DOI: 10.1139/cjm-2014-0016
  • Peer Reviewed
• [Journal Article] Identification of leptospiral 3-hydroxyacyl-CoA dehydrogenase released in the urine of infected hamsters. (2014)
  • Author(s): Segawa T, Nomura HK, Villanueva SY, Saito M, Nomura KI, Gloriani NG, Yoshida S
  • Journal Title: BMC Micrbiol Volume: 14 Issue: 1 Pages: 132-141
  • DOI: 10.1186/1471-2180-14-132
  • Peer Reviewed
• [Journal Article] Leptospira-rat-human relationship in Luzon, Philippines. (2014)
  • Author(s): Villanueva SY, Saito M, Baterna RA, Estrada CA, Rivera AK, Dato MC, Zamora PR, Segawa T, Cavinta LL, Fukui T, Masuzawa T, Yanagihara Y, Gloriani NG, Yoshida S
  • Journal Title: Microbes Infect Volume: 16 Issue: 11 Pages: 902-910
  • DOI: 10.1016/j.micinf.2014.07.001
  • Peer Reviewed
• [Journal Article] PCR and culture identification of pathogenic Leptospira spp. from coastal soil in Leyte, Philippines after a storm surge during Super Typhoon Haiyan (Yolanda). (2014)
  • Author(s): Saito M, Miyahara S, Villanueva SY, Aramaki N, Ikejiri M, Kobayashi Y, Guevarra JP, Masuzawa T, Gloriani NG, Yanagihara Y, Yoshida S
  • Journal Title: Appl Environ Microb Volume: 80 Issue: 22 Pages: 6926-6932
  • DOI: 10.1128/aem.02568-14
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] レプトスピラ感染症 ～ワイル病病原体発見から百年～ (2014)
  • Author(s): 齋藤光正、Sharon Y.A.M. Villanueva、増澤俊幸、柳原保武、吉田眞一
  • Journal Title: 日本細菌学雑誌 Volume: 69 Pages: 589-600
  • NAID: 130004713723
  • Open Access / Acknowledgement Compliant
• [Journal Article] High virulence in hamsters of four dominantly prevailing Leptospira serovars isolated from rats in the Philippines. (2014)
  • Author(s): Villanueva SY, Saito M, Tsutsumi Y, Segawa T, Baterna RA, Chakraborty A, Asoh T, Miyahara S, Yanagihara Y, Cavinta LL, Gloriani NG, Yoshida SI.
  • Journal Title: Microbiology Volume: 160 Issue: 2 Pages: 418-428
  • DOI: 10.1099/mic.0.072439-0
  • Peer Reviewed
• [Journal Article] Leptospira idonii sp. nov., isolated from an environmental water in Fukuoka, Japan. (2013)
  • Author(s): Saito M, Villanueva SY, Kawamura Y, Iida K, Tomida J, Kanemaru T, Kohno E, Miyahara S, Umeda A, Amako K, Gloriani NG, Yoshida SI
  • Journal Title: Int J Syst Evol Microbiol Volume: 63 Issue: Pt_7 Pages: 2457-2462
  • DOI: 10.1099/ijs.0.047233-0
  • Peer Reviewed
• [Journal Article] Comparative analysis of Leptospira strains isolated from environmental soil and water in the Philippines and Japan. (2013)
  • Author(s): Saito M, Villanueva SY, Chakraborty A, Miyahara S, Segawa T, Asoh T, Ozuru R, Gloriani NG, Yanagihara Y, Yoshida SI
  • Journal Title: Appl Environ Microbiol Volume: 79 Issue: 2 Pages: 601-609
  • DOI: 10.1128/aem.02728-12
  • Peer Reviewed
• [Journal Article] Development of immunochromatography-based methods for detection of leptospiral lipopolysaccharide antigen in urine. (2013)
  • Author(s): Widiyanti D, Koizumi N, Fukui T, Muslich LT, Segawa T, Villanueva SY, Saito M, Masuzawa T, Gloriani NG, Yoshida SI
  • Journal Title: Clin Vaccine Immunol Volume: 20 Issue: 5 Pages: 683-690
  • DOI: 10.1128/cvi.00756-12
  • Peer Reviewed
• [Presentation] レプトスピラ感染初期における定着・増殖の場の形態学的探索 (2016)
  • Author(s): 尾鶴亮, 齋藤光正, 金丸孝昭, 吉田眞一
  • Organizer: 第53回レプトスピラ・シンポジウム
  • Place of Presentation: 大阪大学吹田キャンパス（大阪府・吹田市）
  • Year and Date: 2016-03-22
• [Presentation] 忍び寄るレプトスピラ感染症 ―病原体発見の歴史から最新知見まで― (2015)
  • Author(s): 齋藤光正、Sharon Y.A.M. Villanueva、増澤俊幸、柳原保武、吉田眞一
  • Organizer: 第85回日本感染症学会西日本地方会学術集会
  • Place of Presentation: 奈良春日野国際フォーラム（奈良県・奈良市）
  • Year and Date: 2015-10-16
  • Invited
• [Presentation] 台風Haiyanによる高潮被害を受けたフィリピン・レイテ島の土壌からの病原性レプトスピラ分離 (2015)
  • Author(s): 宮原 敏、齋藤光正、Villanueva SY、荒牧夏美、池尻真美、小林好江、Guevarra JP、増澤俊幸、Gloriani NG、柳原保武、吉田眞一
  • Organizer: 第52回レプトスピラ・シンポジウム
  • Place of Presentation: 岐阜
  • Year and Date: 2015-03-28
• [Presentation] Development and evaluation of vaccines using 5 Leptospira serovars prevailing in the Philippines. (2015)
  • Author(s): Villanueva SY, Yanagihara Y, Saito M, Baterna RA, Estrada CA, Rivera AK, Segawa T, Ozuru R, Ikejiri M, Hidaka Y,Wang C, Gloriani NG, Yoshida S
  • Organizer: 第52回レプトスピラ・シンポジウム
  • Place of Presentation: 岐阜
  • Year and Date: 2015-03-28
• [Presentation] レプトスピラの生態に関する新たな視点 (2015)
  • Author(s): 柳原保武、齋藤光正、増澤俊幸、Villanueva SY、宮原 敏、小林好江、日高悠介、荒牧夏美、池尻真美、Baterna RA、Baterna RA、Mingala BX、Guevarra JP、Gloriani NG、吉田真一
  • Organizer: 第52回レプトスピラ・シンポジウム
  • Place of Presentation: 岐阜
  • Year and Date: 2015-03-28
• [Presentation] Luciferase 発現 Leptospira を用いた in vivo イメージング (2015)
  • Author(s): 尾鶴 亮、齋藤光正、吉田眞一
  • Organizer: 第52回レプトスピラ・シンポジウム
  • Place of Presentation: 岐阜
  • Year and Date: 2015-03-28
• [Presentation] レプトスピラ感染症における抗Patoc抗体出現の意味 (2015)
  • Author(s): 池尻真美、Villanueva SY、齋藤光正、吉開泰信、吉田眞一
  • Organizer: 第52回レプトスピラ・シンポジウム
  • Place of Presentation: 岐阜
  • Year and Date: 2015-03-28
• [Presentation] 環境分離レプトスピラ株の病原性の検討 (2015)
  • Author(s): 日高悠介、齋藤光正、Villanueva SY、増澤俊幸、金丸孝昭、柳原保武、Gloriani NG，吉田眞一
  • Organizer: 第52回レプトスピラ・シンポジウム
  • Place of Presentation: 岐阜
  • Year and Date: 2015-03-28
• [Presentation] 日本全国の土壌，環境水からのレプトスピラ分離株の性状解析 (2015)
  • Author(s): 増澤俊幸、縁川むつみ、工藤芳子、齋藤光正、Villanueva SY、岡本能弘、福井貴史、柳原保武、吉田真一
  • Organizer: 第52回レプトスピラ・シンポジウム
  • Place of Presentation: 岐阜
  • Year and Date: 2015-03-28
• [Presentation] 半固形コルトフ培地を用いた福岡都心部の野生ラットからの病原性レプトスピラ分離と分離株の解析 (2015)
  • Author(s): 齋藤光正、Villanueva SY、増澤俊幸、宮原 敏、尾鶴 亮、山口孝治、吉村芳修、池尻真美、荒牧夏美、飯田健一郎、柳原保武、吉田眞一
  • Organizer: 第88回日本細菌学会
  • Place of Presentation: 岐阜
  • Year and Date: 2015-03-26 – 2015-03-28
• [Presentation] 土壌、水などの環境からのレプトスピラ分離と性状解析 (2015)
  • Author(s): 増澤俊幸、齋藤光正、Villanueva SY、岡本能弘、福井貴史、柳原保武、吉田眞一
  • Organizer: 第88回日本細菌学会
  • Place of Presentation: 岐阜
  • Year and Date: 2015-03-26 – 2015-03-28
• [Presentation] フィリピンで流行している5つのレプトスピラの血清型に対するワクチンの開発と評価 (2015)
  • Author(s): Villanueva SY、柳原保武、齋藤光正、瀬川孝耶、尾鶴 亮、池尻真美、Gloriani NG、吉田眞一
  • Organizer: 第88回日本細菌学会
  • Place of Presentation: 岐阜
  • Year and Date: 2015-03-26 – 2015-03-28
• [Presentation] New knowledges about Leptospira strains isolated from environmental water and soil by using a novel combination of selective agents. (2014)
  • Author(s): Saito M
  • Organizer: 3rd Scientific Symposium on the Prevention and Control of Leptospirosis in the Philippines
  • Place of Presentation: Manila, Philippines
  • Year and Date: 2014-12-02
  • Invited
• [Presentation] 新生仔ラットのレプトスピラ感受性/抵抗性に関する検討 (2014)
  • Author(s): 齋藤光正、Villanueva SY、吉田眞一
  • Organizer: 第88回日本感染症学会学術集会
  • Place of Presentation: 福岡
  • Year and Date: 2014-06-18 – 2014-06-20
• [Presentation] New insights in the study of Leptospira in the environment using a novel combination of selective agents. (2013)
  • Author(s): Saito M, Villanueva SY, Iida K, Chakraborty A, Miyahara S, Segawa T, Asoh T, Ozuru R, Kawamura Y, Tomida J, Kohno E, Umeda A, Gloriani NG, Yanagihara Y, Yoshida SI
  • Organizer: 8th Scientific Meeting of International Leptospirosis Society, October 8-11, 2013, Fukuoka.
  • Place of Presentation: 福岡
• [Presentation] Ripping the hepatocyte junction apart by intercellular invasion of Leptospira causes jaundice in a hamster model of Weil’s disease. (2013)
  • Author(s): Miyahara S, Saito M, Kanemaru T, Villanueva SY, Gloriani NG, Yoshida SI
  • Organizer: 8th Scientific Meeting of International Leptospirosis Society
  • Place of Presentation: 福岡
• [Presentation] A survey on the prevailing Leptospira serovars among humans and animals in the Philippines, August 2010 to February 2012. (2013)
  • Author(s): Gloriani NG, Cavinta LL, Baterna RA, Estrada CAM, Rivera AKB, Dato MC, Villanueva CYAM, Saito M, Yanagihara Y, Yoshida SI
  • Organizer: 8th Scientific Meeting of International Leptospirosis Society
  • Place of Presentation: 福岡
• [Presentation] Leptospira isolates from humans and rats in the Philippines: insights into the Leptospira-rat-human relationship (2013)
  • Author(s): Villanueva SYAM, Saito M, Baterna RA, Estrada CAM, Rivera AKB, Dato MC, Segawa T, Cavinta LL, Fukui T, Masuzawa T, Yanagihara Y, Gloriani NG, Yoshida SI
  • Organizer: 8th Scientific Meeting of International Leptospirosis Society
  • Place of Presentation: 福岡
• [Presentation] Identification of leptospiral 3-hydroxyacyl-CoA dehydrogenase secreted in the urine of infected hamsters (2013)
  • Author(s): Segawa T, Nomura K, Villanueva SYAM, Saito M, Nomura K, Gloriani NG, Yoshida SI
  • Organizer: 8th Scientific Meeting of International Leptospirosis Society
  • Place of Presentation: 福岡
• [Presentation] Natural defense by saliva, mucosa, and gastric acid against oral infection by Leptospira. (2013)
  • Author(s): Asoh T, Saito M, Villanueva SYAM, Kanemaru T, Gloriani NG, Yoshida SI
  • Organizer: 8th Scientific Meeting of International Leptospirosis Society
  • Place of Presentation: 福岡
• [Presentation] Novel immunochromatography-based methods for detection of leptospiral antigen in human urine. (2013)
  • Author(s): Widiyanti D, Nobuo K, Fukui T, Muslich LT, Segawa T, Villanueva SYAM, Saito M, Masuzawa T, Gloriani NG, Yoshida SI
  • Organizer: 8th Scientific Meeting of International Leptospirosis Society
  • Place of Presentation: 福岡
• [Presentation] Characterization of Leptospira infection in suckling and weaning rat pups. (2013)
  • Author(s): Muslich LT, Villanueva SYAM, Amran MY, Segawa T, Saito M, Yoshida SI
  • Organizer: 8th Scientific Meeting of International Leptospirosis Society
  • Place of Presentation: 福岡
• [Presentation] レプトスピラ感染症 ～ワイル病病原体発見から百年～
  • Author(s): 齋藤光正、宮原 敏、金丸孝昭、吉田眞一
  • Organizer: 第26回九州電子顕微鏡技術研究会
  • Place of Presentation: 福岡
  • Invited
• [Presentation] ５種類の抗菌薬を選択剤として用いた環境中レプトスピラの分離と分離株の解析
  • Author(s): 齋藤光正、Sharon Y.A.M. Villanueva、麻生達磨、河村好章、柳原保武、吉田眞一
  • Organizer: 第87回日本感染症学会
  • Place of Presentation: 横浜
• [Book] 「レプトスピラ感染症」小児疾患診療のための病態整理１第５版 (2014)
  • Author(s): 齋藤光正
  • Total Pages: 1500
  • Publisher: 東京医学社
• [Book] 戸田新細菌学（改訂34版）「細菌の生理・生化学」 (2013)
  • Author(s): 齋藤光正（分担執筆）
  • Total Pages: 55
  • Publisher: 南山堂