1987 Fiscal Year Final Research Report Summary
Characteristic antigen-determinants of pathogenic microorganisms and platelet
| Project/Area Number |
61480180
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
内科学一般
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| Research Institution | Ehime University |
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Principal Investigator |
KOBAYASHI Yuzuru Ehime University, Professor, 医学部, 教授 (20038615)
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| Co-Investigator(Kenkyū-buntansha) |
KANEMITSU Nozomu Ehime University, Assistant, 医学部附属病院, 助手 (20177546)
HATO Takaaki Ehime University, Assistant, 医学部, 助手 (30172943)
TAMAI Tomonori Ehime University, Assistant, 医学部, 助手 (20179873)
YASUKAWA Masaki Ehime University, Instructor, 医学部附属病院, 講師 (60127917)
SIOZAKA Takahiko Ehime University, Instructor, 医学部府属病院, 講師 (90035486)
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| Project Period (FY) |
1986 – 1987
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| Keywords | Monoclonal antibody / Japanese encephalitis virus / Weil's disease / Leptospira / Tsutsugamushi disease / Rickettsia tsutsugamushi / Platelet / CD9抗原 |
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| Research Abstract |
1. Antigenic relationships among Japanese encephalitis (JE) virus strains were analyzed by using monoclonal antibodies against Nakayama and Kamiyama strains in the hemagglutination inhibition and neutralization tests. The strains of JE virus were classified into four antigenic groups. Immunoblotting analysis and competitive binding assay were performed to determine the distribution of antigenic determinants. The results suggest the existence of at least five HI sites on the JE virus, and indicated that the JE species-specific HI site and the flavivirus genus-specific HI site are topologically distinct.
2. Monoclonal antibodies against etiological agents of Weil's disease were produced. Identification using these highly specific antibodies are extremely useful for laboratory diagnosis of the disease. Leptospires of serogroup Icterohaemorrhagiae were serologically analyzed using monoclonal antibodies against serovars icterohaemorrhagiae, copenhageni, smithi and naam. They were divided int
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o the three subgroups. Restriction endonuclease analysis for the classification of leptospires were also performed.
3. Monoclonal antibodies against Gilliam, Karp, Kato, and newly isolated two other strains of Rickettsia tsutsugamushi were produced. Antigenic relationships among these strains and many other strains were analyzed by using the antibodies. Each strain has type specific antigens and their common antigens. SDS-PAGE and immunoblot of antigens from the rickettsial strains against the monoclonal antibodies revealed that molecular sizes of type specific antigens were 60kD and those of common antigens were 44kD, 60kD and 61kD.
4. A monoclonal antibody to CD9 antigen, PMA2, induced fibrinogen binding to platelets. The binding of ^<125>I-fibrinogen to platelets occurred in a PMA2 concentration dependent manner. Combined treatment with CP/CPK and aspirin abolished fibrinogen binding and aggregation. This result demonstrated that the binding of IgG molecules to the CD9 antigen exposes fibrinogen receptors via both secreted ADP and thromboxane and that either one of both can expose the receptors to an extent sufficient to aggregate platelets. Less
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