質量分析計によるトキソプラズマ症の再活性化検出法及び重症度の評価法の確立

2018 Fiscal Year Research-status Report

• Project/Area Number: 17KT0124
• Research Institution: Chiba University
• Principal Investigator: 野呂瀬 一美 千葉大学, 大学院医学研究院, 特任准教授 (30156244)
• Co-Investigator(Kenkyū-buntansha): 彦坂 健児 千葉大学, 大学院医学研究院, 講師 (30456933) ; 佐藤 守 千葉大学, 医学部附属病院, 特任准教授 (20401002) ; 野村 文夫 千葉大学, 医学部附属病院, 特任教授 (80164739)
• Project Period (FY): 2017-07-18 – 2020-03-31
• Keywords: トキソプラズマ / ステージ変換 / 再活性化

Outline of Annual Research Achievements

細胞内寄生体トキソプラズマの人体内における発育ステージとしては急増虫体のタキゾイトとシストを形成する緩増虫体のブラディゾイトの２種類がある。それらは病原性や再発に密接に関係しているため虫体の発育分化に影響を与える因子や機構を解析することはトキソプラズマ症の制圧に非常に重要である。
昨年度は、in vitroにおけるタキゾイトからシスト形成、及び逆方向であるシストからタキゾイトへの活性化について培養条件を変化させて観察した。
今年度は、培養上清中のタンパク質の変化を質量分析で解析する予定であったが、当初GKOマウスの繁殖に問題があり、胎児繊維芽細胞が得にくかったため、in vitroの実験が遅れていたが、秋以降GKOマウスの繁殖に成功したため、実験を再開している。
臨床検体は今年度は39検体受け入れ、トキソプラズマのPCR診断を行った。それらの臨床経過より重症度を分類化した。さらに、過去PCR陽性であった症例の重症度を分類化し、現在、それらの血清タンパク質の質量分析に向けて準備段階にある。

Current Status of Research Progress

3: Progress in research has been slightly delayed.

Reason

GKOマウスの維持に問題があり、野生型マウスと交配させることにより、新たにGKOマウス株を樹立させている。そのため、一時期,胎児繊維芽細胞を得ることが困難になっていた。

Strategy for Future Research Activity

2019年度は、様々な条件下での培養上清中のタンパク質の変化を質量分析法を用いたプロテオーム解析により同定する。
また、臨床検体の血清のプロテオーム解析を行い、重症度の分類と照らし合わせて、再活性化の検出法を確立する。

Research Products

• [Journal Article] Anti-FIRΔexon2, a splicing variant form of PUF60, auto-antibody is detected in the sera of esophageal squamous cell carcinoma. (2019)
  • Author(s): Kobayashi S, Hiwasa T, Ishige T, Rahmutulla B, Kano M, Hoshino T, Minamoto T, Nomura F, Matsubara H, Matsushita K
  • Journal Title: Cancer Sci. Volume: 110 Pages: 2004-2013
  • DOI: 10.1111/cas.14024
  • Peer Reviewed / Open Access
• [Journal Article] Identification of conjugation positions of urinary glucuronidated vitamin D3 metabolites by LC/ESI-MS/MS after conversion to MS/MS-fragmentable derivatives. (2019)
  • Author(s): Yoshimura Y, Hibi R, Nakata A, Togashi M, Ogawa S, Ishige T, Satoh M, Nomura F, Higashi T
  • Journal Title: Biomed Chromatogr. Volume: 33 Pages: e4538
  • DOI: 10.1002/bmc.4538
  • Peer Reviewed
• [Journal Article] 原発性脳リンパ腫を合併したトキソプラズマ脳炎の再発例 (2019)
  • Author(s): 鷲野 巧弥, 矢嶋 敬史郎, 福島 一彰, 佐々木 秀悟, 関谷 綾子, 関谷 紀貴, 柳澤 如樹, 野呂瀬 一美, 味澤 篤, 今村 顕史.
  • Journal Title: 感染症学雑誌 Volume: - Pages: -
  • Peer Reviewed
• [Journal Article] Metastatic Malignant Lymphoma Mimicking Cerebral Toxoplasmosis with the "Target Sign" (2019)
  • Author(s): Ueno H, Norose K, Kamimura T, Mihara K, Yamasaki F, Hikosaka K, J Amatya V, Takeshima Y, Kurisu K, Maruyama H
  • Journal Title: Intern Med Volume: 58 Pages: 1157-1162
  • DOI: 10.2169/internalmedicine.1156-18
  • Peer Reviewed / Open Access
• [Journal Article] Infection dynamics of Toxoplasma gondii in gut-associated tissues after oral infection: The role of Peyer's patches. (2019)
  • Author(s): Mitsunaga T, Norose K, Aosai F, Horie H, Ohnuma N, Yano A
  • Journal Title: Parasitol Int Volume: 68 Pages: 40-47
  • DOI: 10.1016/j.parint.2018.08.010
  • Peer Reviewed
• [Journal Article] Disseminated toxoplasmosis with atypical symptoms which developed with exacerbation of systemic lupus erythematosus. (2019)
  • Author(s): Furuya H, Ikeda K, Iida K, Suzuki K, Furuta S, Tamachi T, Suzuki K, Miura G, Hiraguri M, Hase R, Hikosaka K, Norose K, Nakajima H
  • Journal Title: Lupus Volume: 28 Pages: 133-136
  • DOI: 10.1177/0961203318815583
  • Peer Reviewed
• [Journal Article] Identification of specific and common diagnostic antibody markers for gastrointestinal cancers by SEREX screening using testis cDNA phage library. (2018)
  • Author(s): Kobayashi S, Hiwasa T, Arasawa T, Kagaya A, Ishii S, Shimada H, Ito M, Suzuki M, Kano M, Rahmutulla B, Kitamura K, Sawabe Y, Shin H, Takiguchi M, Nomura F, Matsubara H, Matsushita K.
  • Journal Title: Oncotarget Volume: 9 Pages: 18559-18569
  • DOI: 10.18632/oncotarget.24963
  • Peer Reviewed / Open Access
• [Journal Article] One novel and two uncommon MEFV mutations in Japanese patients with familial Mediterranean fever: a clinicogenetic study. (2018)
  • Author(s): Kishida D, Yazaki M, Nakamura A, Nomura F, Kondo T, Uehara T, Ikusaka M, Ohya A, Watanabe N, Endo R, Kawaai S, Shimojima Y, Sekijima Y.
  • Journal Title: Rheumatol Int Volume: 38 Pages: 105-110
  • DOI: 10.1007/s00296-017-3886-z
  • Peer Reviewed / Open Access
• [Journal Article] Accelerated oligosaccharide absorption and altered serum metabolites during oral glucose tolerance test in young Japanese with impaired glucose tolerance. (2018)
  • Author(s): Miki T, Lee EY, Eguchi A, Sakurai K, Sawabe Y, Yoshida T, Saito K, Yokoh H, Ishikawa K, Yokote K, Kuzuya T, Miki E, Mori C, Nomura F.
  • Journal Title: J Diabetes Investig Volume: 9 Pages: 512-521
  • DOI: 10.1111/jdi.12719
  • Peer Reviewed / Open Access
• [Journal Article] Disturbed alternative splicing of FIR (PUF60) directed cyclin E overexpression in esophageal cancers. (2018)
  • Author(s): Ogura Y, Hoshino T, Tanaka N, Ailiken G, Kobayashi S, Kitamura K, Rahmutulla B, Kano M, Murakami K, Akutsu Y, Nomura F, Itoga S, Matsubara H, Matsushita K.
  • Journal Title: Oncotarget Volume: 9 Pages: 22929-22944
  • DOI: 10.18632/oncotarget.25149
  • Peer Reviewed / Open Access
• [Journal Article] Determination of serum carbohydrate-deficient transferrin by a nephelometric immunoassay for differential diagnosis of alcoholic and non-alcoholic liver diseases. (2018)
  • Author(s): Nomura F, Kanda T, Seimiya M, Satoh M, Kageyama Y, Yamashita T, Yokosuka O, Kato N, Maruyama K.
  • Journal Title: Clin Chim Acta Volume: 485 Pages: 181-186
  • DOI: 10.1016/j.cca.2018.06.040
  • Peer Reviewed / Open Access
• [Journal Article] Search for a Novel Allergen in Hen's Egg Allergy Using an IgE Immunoblotting Assay. (2018)
  • Author(s): Sogawa K, Takahashi Y, Shibata Y, Satoh M, Kodera Y, Nomura F, Tanaka T, Sato H, Yamaide F, Nakano T, Iwahashi K, Sugita-Konishi Y, Shimada A, Shimojo N.
  • Journal Title: Int Arch Allergy Immunol Volume: 176 Pages: 189-197
  • DOI: 10.1159/000488144
  • Peer Reviewed
• [Journal Article] Capsular serotyping of Haemophilus influenzae by using matrix-associated laser desorption ionization-time of flight mass spectrometry. (2018)
  • Author(s): Takeuchi N, Segawa S, Ishiwada N, Ohkusu M, Tsuchida S, Satoh M, Matsushita K, Nomura F
  • Journal Title: J Infect Chemother Volume: 24 Pages: 510-514
  • DOI: 10.1016/j.jiac.2018.02.007
  • Peer Reviewed / Open Access
• [Journal Article] Application of the biocopolymer preparation system, rapid BACpro® II kit, for mass-spectrometry-based bacterial identification from positive blood culture bottles by the MALDI Biotyper system. (2018)
  • Author(s): Tsuchida S, Murata S, Miyabe A, Satoh M, Takiwaki M, Ashizawa K, Terada T, Ito D, Matsushita K, Nomura F
  • Journal Title: J Microbiol Methods Volume: 152 Pages: 86-91
  • DOI: 10.1016/j.mimet.2018.07.017
  • Peer Reviewed / Open Access
• [Journal Article] Surveillance evaluation of the standardization of assay values for serum total 25-hydroxyvitamin D concentration in Japan. (2018)
  • Author(s): Ihara H, Kiuchi S, Ishige T, Nishimura M, Matsushita K, Satoh M, Nomura F, Yamashita M, Kitajima I, Tsugawa N, Okano T, Hirota K, Miura M, Totani M, Hashizume N.
  • Journal Title: Ann Clin Biochem Volume: 55 Pages: 647-656
  • DOI: 10.1177/0004563218765570
  • Peer Reviewed
• [Journal Article] High-throughput genotyping of GC (vitamin D-binding protein) by melting analysis with locked nucleic acid-incorporating dual hybridization probe for improving mismatch discrimination. (2018)
  • Author(s): Ishige T, Satoh M, Itoga S, Nishimura M, Matsushita K, Nomura F
  • Journal Title: Clin Chim Acta Volume: 487 Pages: 126-132
  • DOI: 10.1016/j.cca.2018.09.034
  • Peer Reviewed / Open Access
• [Journal Article] Current Status of Proteomic Technologies for Discovering and Identifying Gingival Crevicular Fluid Biomarkers for Periodontal Disease. (2018)
  • Author(s): Tsuchida S, Satoh M, Takiwaki M, Nomura F
  • Journal Title: Int J Mol Sci Volume: 20 Pages: 86
  • DOI: 10.3390/ijms20010086
  • Peer Reviewed / Open Access
• [Presentation] 多摩川河口域に飛来する野鳥の腸管寄生虫感染状況の解明 (2019)
  • Author(s): 彦坂健児、田中咲帆、馮 雪、篠田渉平、LiLi、Li Kexin, 野呂瀬 一美
  • Organizer: 第８８回日本寄生虫学会大会
• [Remarks] 感染生体防御学
  • URL: http://www.m.chiba-u.ac.jp/class/infection-hostdefense/