2005 Fiscal Year Final Research Report Summary
IMMUNE RESPONSE TO GLYCOLIPIDS SEPARETED FROM MYCOBACTERIUM TUBERCULOSIS IN GUINEA PIG
| Project/Area Number |
16590369
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Bacteriology (including Mycology)
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| Research Institution | KITASATO UNIVERSITY |
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Principal Investigator |
KUMAZAWA Yoshio KITASATO UNIV., SCHOOL OF SCIENCE, PROFESSOR, 理学部, 教授 (30072375)
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| Co-Investigator(Kenkyū-buntansha) |
TAKIMOTO Hiroaki KITASATO UNIV., SCHOOL OF SCIENCE, LECTURER, 理学部, 講師 (00253534)
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| Project Period (FY) |
2004 – 2005
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| Keywords | guinea pigs / mycobacterial glycolipid / CD1 / cytokine / IL-12 / IFN-γ / monoclonal antibody |
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| Research Abstract |
1)Chronic inflammation is the hallmark of pathogenesis of tuberculosis and due to the immune responses to not only protein antigens but also various glycolipids produced from Mycobacterium tuberculosis. The mechanisms by which mycobacterial lipid antigens present to T cells via CD1 family, especially CD1b molecule, have been yet unclear. Since guinea pigs carry CD1b molecule, we compared immune responses to protein and glycolipid antigens in guinea pigs. Hartley guinea pigs were immunized with a crude lipid fraction (CLF) separated from M.tuberculosis H37Rv, ovalbumin (OVA) and self-made recombinant CFP-10 emulsified with muramyl dipeptide and incomplete Freund's adjuvant. A control group was immunized similarly with OVA and CFP-10 without CLF. As expected, immune responses to CLF were observed only in the immunized group, and significant CLF-specific delayed-type hypersensitivity (DTH) reaction and Th1-type lgG2 antibody response were demonstrated. CLF consisted of caldiolipin, phosphatidylinositol mannosides (PIMs), trehalose monomycolate (TMM), trehalose dimycolate (TDM) etc. CLF-specific proliferative T cell responses were also demonstrated.
2)Recombinant guinea pig (gp) IL-12 p70 and IFN-g were produced by using Sf9 cells transfected with baculo virus AcRP23-LacZ carrying gpIFN-γ, or gpIL-12p40 and gpIL-12p35 genes. We obtained biologically active rgpIFN-γ and rgpIL-12 molecules. Using purified these proteins, mice were immunized for preparing monoclonal antibodies (mAb). We got several mAb clones to rgpIFN-γ and rgpIL-12 which were able to neutralize the cytokine activy.
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