Establishment of antibody-cocktail therapy against invading bacteria for sepsis patients

• Project/Area Number: 18K08868
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 55050:Anesthesiology-related
• Research Institution: Tokai University
• Principal Investigator: Watanabe Nobuo 東海大学, 医学部, 助教 (80396928)
• Co-Investigator(Kenkyū-buntansha): 井上 茂亮 東海大学, 医学部, 准教授 (30582209)
• Project Period (FY): 2018-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2020: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2019: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2018: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: モノクローナル抗体 / 感染症 / 薬剤耐性 / 活性抗体 / 膜タンパク質 / 抗菌活性 / ペプチドグリカン / クオラムセンシング / 多剤耐性菌 / MRSA / 感染菌 / 抗体

Outline of Final Research Achievements

Establishment of a therapeutic strategy against infectious diseases caused by drug-resistant bacteria including Methicillin-Resistant Staphylococcus aureus (MRSA) is a long-standing challenge; however, almost no antibody to MRSA has successfully showed effectiveness in the clinical trials. In this study, we targeted 2 master proteins responsible for toxin synthesis and 3 key proteins responsible for cell wall biosynthesis, respectively, and generated a total of 25 monoclonal antibodies in mice. Unfortunately, however, none of them demonstrated expected anti-MRSA activity on its own. However, now it has become possible to assess the anti-MRSA effect of these antibodies in combination with other antibodies and antibiotics etc.

Academic Significance and Societal Importance of the Research Achievements

本研究において作製した抗MRSA抗体はいずれも単独では当初期待した効果(抗毒素産生作用ならびに抗増殖作用)は見られなかった。しかしこの結果、5種のタンパク質に関しては、今後の標的とすべき配列が絞り込まれたことになる。また、今後は他の抗体ならびに各種の抗菌剤との併用効果を検証しすることによって付加的な抗菌効果が得られるか検討する予定である。ハイブリドーマは樹立され液体窒素に保管されているため、必要に応じてすぐに抗体の追加産生ならびにヒト化抗体化が可能である。

Research Products

• [Journal Article] Usefulness of next-generation DNA sequencing for the diagnosis of urinary tract infection (2020)
  • Author(s): Ishihara T, Watanabe N, Inoue S, Aoki H, Tsuji T, Yamamoto B, Yanagi H, Oki M1, Kryukov K, Nakagawa S, Inokuchi S, Ozawa H, Imanishi T
  • Journal Title: Drug Discoveries & Therapeutics Volume: 14 Issue: 1 Pages: 42-49
  • DOI: 10.5582/ddt.2020.01000
  • NAID: 130007802636
  • ISSN: 1881-7831, 1881-784X
  • Year and Date: 2020-02-29
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Podoplanin is indispensable for cell motility and platelet-induced epithelial-to-mesenchymal transition-related gene expression in esophagus squamous carcinoma TE11A cells (2020)
  • Author(s): Watanabe N, Kidokoro M, Tanaka M, Inoue S, Tsuji T, Akatuska H, Okada C, Iida Y, Okada Y, Suzuki Y, Sato T, Yahata T, Hirayama N, Nakagawa Y, Inokuchi S.
  • Journal Title: Cancer Cell International Volume: 20 Issue: 1 Pages: 263-263
  • DOI: 10.1186/s12935-020-01328-2
  • Peer Reviewed / Open Access
• [Journal Article] A pull-down and slot blot-based screening system for inhibitor compounds of the podoplanin-CLEC-2 interaction (2019)
  • Author(s): Watanabe N, Kidokoro M, Suzuki Y, Tanaka M, Inoue S, Tsukamoto H, Hirayama N, Hsieh PW, Tseng CP, Nakagawa Y, Inokuchi S
  • Journal Title: PLOS ONE Volume: 14 Issue: 9 Pages: e0222331-e0222331
  • DOI: 10.1371/journal.pone.0222331
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Papaverine identified as an inhibitor of high mobility group box 1/receptor for advanced glycation end-products interaction suppresses high mobility group box 1-mediated inflammatory responses. (2019)
  • Author(s): Tamada K, Nakajima S, Ogawa N, Inada M, Shibasaki H, Sato A, Takasawa R, Yoshimori A, Suzuki Y, Watanabe N, Oyama T, Abe H, Inoue S, Abe T, Yokomizo T, Tanuma S.
  • Journal Title: Biochem Biophys Res Commun Volume: 511 Issue: 3 Pages: 665-670
  • DOI: 10.1016/j.bbrc.2019.01.136
  • Peer Reviewed / Open Access
• [Journal Article] Detection of pathogenic bacteria in the blood from sepsis patients using 16S rRNA gene amplicon sequencing analysis. (2018)
  • Author(s): Watanabe N, Kryukov K, Nakagawa S, Takeuchi JS, Takeshita M, Kirimura Y, Mitsuhashi S, Ishihara T, Aoki H, Inokuchi S, Imanishi T, Inoue S
  • Journal Title: PLoS One Volume: 13 Issue: 8 Pages: e0202049-e0202049
  • DOI: 10.1371/journal.pone.0202049
  • NAID: 120006826803
  • Peer Reviewed / Open Access / Int'l Joint Research