Faculty Profiles - Uchiyama Jumpei

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黄色ブドウ球菌ファージS13′およびS24‐1ゲノム比較による尾部吸着タンパク質の同定と解析

内山淳平, 竹村伊代, 佐藤美帆, 加藤伸一郎, 氏原隆子, 松崎茂展, 大畑雅典

日本ブドウ球菌研究会プログラム・講演要旨集 56th 18 2011

バクテリオファージによるマウス緑膿菌角膜炎の抑制効果

福田憲, 石田わか, 角環, 森田珠恵, 内山淳平, 松崎茂展, 大畑雅典, 福島敦樹

日本眼感染症学会・日本眼炎症学会・日本コンタクトレンズ学会総会プログラム・講演抄録集 48th-45th-54th 51 2011

Isolation and characterization of a novel Staphylococcus aureus bacteriophage, phi MR25, and its therapeutic potential

Hiroshi Hoshiba, Jumpei Uchiyama, Shin-ichiro Kato, Takako Ujihara, Asako Muraoka, Masanori Daibata, Hiroshi Wakiguchi, Shigenobu Matsuzaki

ARCHIVES OF VIROLOGY 155 ( 4 ) 545 - 552 2010.4

Language：English Publisher：SPRINGER WIEN

A novel bacteriophage, phi MR25, was isolated from a lysogenic Staphylococcus aureus strain by mitomycin C induction. Its biological features were analyzed in comparison with phi MR11, which was described previously as a prototype therapeutic phage. phi MR25 is morphologically similar to phi MR11 (morphotype B1 of family Myoviridae) but has a broader host range than phi MR11 on S. aureus strains. phi MR25 can also multiply on S. aureus lysogens of phi MR11. Its DNA is 44,342 bp in size, is predicted to include 70 open reading frames, and does not contain genes related to toxin or drug resistance. The lysogenic module and most of the putative virion protein genes are completely different from those of phi MR11. In spite of their genetic diversity, intraperitoneal administration of phi MR25 rescued mice inoculated with a lethal dose of S. aureus, as was the case for phi MR11. These results suggest that phi MR25 could be another candidate phage to treat S. aureus infection.

DOI： 10.1007/s00705-010-0623-2

Web of Science

PubMed

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腸球菌ファージφEF24C由来溶菌酵素ORF9の解析

内山淳平, 林幾江, 竹村伊代, 氏原隆子, 竹内啓晃, 杉浦哲朗, 菅井基之, 大畑雅典, 松崎茂展

日本細菌学雑誌 65 ( 1 ) 180 - 180 2010.2

新規Serratia marcescensバクテリオファージKSP20,KSP90,およびKSP100の性状解析

松下憲司, 内山淳平, 氏原隆子, 杉原重喜, 邑岡麻子, 大畑雅典, 松崎茂展

日本細菌学雑誌 65 ( 1 ) 179 - 179 2010.2

マウスにおける治療用候補バクテリオファージの血中動態の比較検討

内山淳平, 脇口宏, 松崎茂展

感染症学雑誌 83 ( 5 ) 616 - 616 2009.9

Enterococcus faecalisファージφEF24C由来溶菌酵素ORF9の溶菌活性

竹村伊代, 内山淳平, 竹内啓晃, 杉浦哲朗, 松崎茂展

感染症学雑誌 83 ( 5 ) 612 - 613 2009.9

Blood kinetics of four intraperitoneally administered therapeutic candidate bacteriophages in healthy and neutropenic mice

UCHIYAMA Jumpei, MAEDA Yoshihiro, TAKEMURA Iyo, CHESS-WILLIAMS Russ, WAKIGUCHI Hiroshi, MATSUZAKI Shigenobu

Microbiology and immunology 53 ( 5 ) 301 - 304 2009.5

Authorship：Lead author Language：English Publisher：John Wiley & Sons Australia, Ltd

Due to multiple-drug resistant bacteria, phage therapy is being revisited. Although most animal experiments focus on therapeutic efficacy, the blood clearance kinetics of phages have not been well described. For further development of an efficient therapeutic strategy, information on phage blood kinetics is important. In this study, time-course concentration changes in peripheral blood of healthy and neutropenic mice were measured using four therapeutic phages (φMR11, KPP10, φEF24C, and KEP10). The results showed a two- to three-day rapid phage clearance, which fits a two-compartment model. © 2009 The Societies and Blackwell Publishing Asia Pty Ltd.

DOI： 10.1111/j.1348-0421.2009.00125.x

Other Link： http://search.jamas.or.jp/link/ui/2009289589

Enterococcus faecalisファージφEF24C由来溶菌酵素ORF9の溶菌活性

竹村伊代, 内山淳平, 竹内啓晃, 杉浦哲朗, 松崎茂展

感染症学雑誌 83 ( 臨増 ) 256 - 256 2009.3

マウスにおける治療用候補バクテリオファージの血中動態の比較検討

内山淳平, 脇口宏, 松崎茂展

感染症学雑誌 83 ( 臨増 ) 260 - 260 2009.3

緑膿菌の新規溶原性バクテリオファージPAJU2の形態学的・遺伝学的解析

内山淳平, 松崎茂展

日本細菌学雑誌 64 ( 1 ) 141 - 141 2009.2

Tail-associated structural protein gp61 of Staphylococcus aureus phage phi MR11 has bifunctional lytic activity

Mohammad Rashel, Jumpei Uchiyama, Iyo Takemura, Hiroshi Hoshiba, Takako Ujihara, Hiroyoshi Takatsuji, Koichi Honke, Shigenobu Matsuzaki

FEMS MICROBIOLOGY LETTERS 284 ( 1 ) 9 - 16 2008.7

Language：English Publisher：OXFORD UNIV PRESS

A tailed bacteriophage, phi MR11 (siphovirus), was selected as a candidate therapeutic phage against Staphylococcus aureus infections. Gene 61, one of the 67 ORFs identified, is located in the morphogenic module. The gene product (gp61) has lytic domains homologous to CHAP (corresponding to an amidase function) at its N-terminus and lysozyme subfamily 2 (LYZ2) at its C-terminus. Each domain of gp61 was purified as a recombinant protein. Both the amidase [amino acids (aa) 1-150] and the lysozyme (aa 401-624) domains but not the linker domain (aa 151-400) caused efficient lysis of S. aureus. Immunoelectron microscopy localized gp61 to the tail tip of the phi MR11 phage. These data strongly suggest that gp61 is a tail-associated lytic factor involved in local cell-wall degradation, allowing the subsequent injection of phi MR11 DNA into the host cytoplasm. Staphylococcus aureus lysogenized with phi MR11 was also lysed by both proteins. Staphylococcus aureus strains on which phi MR11 phage can only produce spots but not plaques were also lysed by each protein, indicating that gp61 may be involved in 'lysis from without'. This is the first report of the presence of a tail-associated virion protein that acts as a lysin, in an S. aureus phage.

DOI： 10.1111/j.1574-6968.2008.01152.x

In silico and in vivo evaluation of bacteriophage phi EF24C, a candidate for treatment of Enterococcus faecalis infections Reviewed

Jumpei Uchiyama, Mohammad Rashel, Iyo Takemura, Hiroshi Wakiguchi, Shigenobu Matsuzaki

APPLIED AND ENVIRONMENTAL MICROBIOLOGY 74 ( 13 ) 4149 - 4163 2008.7

Authorship：Lead author Language：English Publisher：AMER SOC MICROBIOLOGY

Along with the increasing threat of nosocomial infections by vancomycin-resistant Enterococcus faecalis, bacteriophage (phage) therapy has been expected as an alternative therapy against infectious disease. Although genome information and proof of applicability are prerequisites for a modern therapeutic phage, E. faecalis phage has not been analyzed in terms of these aspects. Previously, we reported a novel virulent phage, phi EF24C, and its biology indicated its therapeutic potential against E. faecalis infection. In this study, the phi EF24C genome was analyzed and the in vivo therapeutic applicability of phi EF24C was also briefly assessed. Its complete genome (142,072 bp) was predicted to have 221 open reading frames (ORFs) and five tRNA genes. In our functional analysis of the ORFs by use of a public database, no proteins undesirable in phage therapy, such as pathogenic and integration-related proteins, were predicted. The noncompetitive directions of replication and transcription and the host-adapted translation of the phage were deduced bioinformatically. Its genomic features indicated that phi EF24C is a member of the SPO1-like phage genus and especially that it has a close relationship to the Listeria phage P100, which is authorized for prophylactic use. Thus, these bioinformatics analyses rationalized the therapeutic eligibility of phi EF24C. Moreover, the in vivo therapeutic potential of phi EF24C, which was effective at a low concentration and was not affected by host sensitivity to the phage, was proven by use of sepsis BALB/c mouse models. Furthermore, no change in mouse lethality was observed under either single or repeated phage exposures. Although further study is required, phi EF24C can be a promising therapeutic phage against E. faecalis infections.

DOI： 10.1128/AEM.02371-07

ファージ療法:現状と展望

松崎茂展, 内山淳平, RASHEL Mohammad, 竹村伊代

日本細菌学雑誌 63 ( 1 ) 56 - 56 2008.2

Revival of Bacteriophage ファージ療法現状と展望

松崎茂展, 内山淳平, Rashel Mohammad, 竹村伊代

日本細菌学雑誌 63 ( 1 ) 56 - 56 2008.2

MEDICAL TOPICS 第26回バクテリオファージを用いる難治性感染症治療

松崎茂展, 内山淳平, RASHEL Mohammad, 竹村伊代

Lung Perspect 16 ( 1 ) 98 - 103 2008.1

MEDICAL TOPICS バクテリオファージを用いる難治性感染症治療

松崎茂展, 内山淳平, Rashel Mohammad, 竹村伊代

THE LUNG-perspectives 16 ( 1 ) 98 - 103 2008.1

Language：Japanese Publisher：(株)メディカルレビュー社

[仮説]ファージ療法は、欧米諸国では抗生物質の実用化に伴い一旦放棄されたが、旧ソ連諸国ポーランドでは現在に至るまで使用されている。ファージ療法の導入により、現在の化学療法の弱点(耐性菌、菌交代症の問題など)を補足することは可能であろうか。[新知見の要点]欧米の多くの研究者によってよく計画された動物実験により、ファージ療法(活性ファージおよびライシンの使用)の有効性・安全性を示すデータが蓄積されつつある。[将来の可能性、問題点]今後、各菌種に対応可能な実用的ファージバンクの構築が必要である。(著者抄録)

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Isolation and characterization of a novel Enterococcus faecalis bacteriophage phi EF24C as a therapeutic candidate Reviewed

Jumpei Uchiyama, Mohammad Rashel, Yoshihiro Maeda, Iyo Takemura, Shigeyoshi Sugihara, Kazue Akechi, Asako Muraoka, Hiroshi Wakiguchi, Shigenobu Matsuzaki

FEMS MICROBIOLOGY LETTERS 278 ( 2 ) 200 - 206 2008.1

Authorship：Lead author Language：English Publisher：BLACKWELL PUBLISHING

Vancomycin-resistant Enterococcus faecalis (VRE) has become a significant threat in nosocomial settings. Bacteriophage (phage) therapy is frequently proposed as a potential alternative therapy for infections caused by this bacterium. To search for candidate therapeutic phages against Enterococcus faecalis infections, 30 Enterococcus faecalis phages were isolated from the environment. One of these, virulent phage phi EF24C, which has a broad host range, was selected for analysis. The plaque-forming ability of phi EF24C was virtually unaffected by differences in the clinical host strains. Furthermore, the phage had a shorter latent period and a larger burst size than ordinary tailed phages, indicating that phi EF24C has effective lytic activity against many Enterococcus faecalis strains, including VRE. Morphological and genomic analyses revealed that phi EF24C is a large myovirus (classified as family Myoviridae morphotype A1) with a linear double-stranded DNA genome of c. 143 kbp. Analyses of the N-terminal amino acid sequences of the virion proteins, together with the morphology and the genome size, speculated that phi EF24C is closely related to other myoviruses of Gram-positive bacteria that have been used experimentally or practically for therapy or prophylaxis. Considering these results, phi EF24C may be a potential candidate therapeutic phage against Enterococcus faecalis infections.

DOI： 10.1111/j.1574-6968.2007.00996.x

Nishikawa, H, Yasuda, M, Uchiyama, J, Rashel, M, Maeda, Y, Takemura, I, Sugihara, S, Ujihara, T, Shimizu, Y, Shuin, T, others

Archives of virology 153 ( 3 ) 507 - 515 2008

Language：English Publisher：Springer

Multidrug-resistant uropathogenic Escherichia coli (UPEC) is increasing gradually on a worldwide scale. We therefore examined the possibility of bacteriophage (phage) therapy for urinary tract infections (UTIs) caused by the UPEC strains as an alternative to chemotherapy. In addition to the well-known T4 phage, KEP10, which was newly isolated, was used as a therapeutic phage candidate. KEP10 showed a broader bacteriolytic spectrum (67%) for UPEC strains than T4 (14%). Morphological and genetic analyses showed that KEP10 resembles phage T4. Phages T4 and KEP10 injected into the peritoneal cavity of mice were distributed immediately to all organs examined and maintained a high titer for at least 24 h. They were stable in the urine of both mice and humans for 24 h at 37 degrees C. Administration of these phages into the peritoneal cavity caused a marked decrease in the mortality of mice inoculated transurethrally with a UPEC strain, whereas most of the control mice died within a few days of bacterial infection. Inoculation with phage alone produced no adverse effects attributable to the phage per se. The present study experimentally demonstrated the therapeutic potential of phage for E. coli-induced UTIs, and T-even-related phages may be suitable candidates with which to treat them.

DOI： 10.1007/s00705-007-0031-4

A novel site-specific recombination system derived from bacteriophage $ϕ$MR11 Reviewed

Rashel, Mohammad, Uchiyama, Jumpei, Ujihara, Takako, Takemura, Iyo, Hoshiba, Hiroshi, Matsuzaki, Shigenobu

Biochemical and biophysical research communications 368 ( 2 ) 192 - 198 2008

Language：English Publisher：Elsevier

We report identification of a novel site-specific DNA recombination system that functions in both it? vivo and ill Vitro, derived from lysogenic Staphylococcus aureus phage phi MR11. In silico analysis of the phi MR11 genome indicated orf1 as a putative integrase gene. Phage and bacterial attachment sites (attP and attB, respectively) and attachment junctions were determined and their nucleotide sequences decoded. Sequences of attP and attB were mostly different to each other except for a two bp common core that was the crossover point. We found several inverted repeats adjacent to the core sequence of attP as potential protein binding sites. The precise and efficient integration properties of phi MR11 integrase were shown on attP and attB in Escherichia coli and the minimum size of attP was found to be 34 bp. In in vitro assays using crude or purified integrase, only buffer and substrate DNAs were required for the recombination reaction, indicating that other bacterially encoded factors are not essential for activity. (C) 2008 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.bbrc.2008.01.045

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