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IMMUNE RESPONSE TO GLYCOLIPIDS SEPARETED FROM MYCOBACTERIUM TUBERCULOSIS IN GUINEA PIG

Research Project

Project/Area Number 16590369
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Bacteriology (including Mycology)
Research InstitutionKITASATO UNIVERSITY

Principal Investigator

KUMAZAWA Yoshio  KITASATO UNIV., SCHOOL OF SCIENCE, PROFESSOR, 理学部, 教授 (30072375)

Co-Investigator(Kenkyū-buntansha) TAKIMOTO Hiroaki  KITASATO UNIV., SCHOOL OF SCIENCE, LECTURER, 理学部, 講師 (00253534)
Project Period (FY) 2004 – 2005
Project Status Completed (Fiscal Year 2005)
Budget Amount *help
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2005: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 2004: ¥1,800,000 (Direct Cost: ¥1,800,000)
Keywordsguinea pigs / mycobacterial glycolipid / CD1 / cytokine / IL-12 / IFN-γ / monoclonal antibody
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.

Report

(3 results)
  • 2005 Annual Research Report   Final Research Report Summary
  • 2004 Annual Research Report

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Published: 2004-04-01   Modified: 2016-04-21  

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