2000 Fiscal Year Final Research Report Summary
Analysis of the interaction between chemokine receptor and V3 region of HIV-gp120 by neutralizing antibodies
| Project/Area Number |
10670418
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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 |
内科学一般
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| Research Institution | Kumamoto University |
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Principal Investigator |
MATSUSHITA Shuzo Center for AIDS Research, Kumamoto University Professor, エイズ学研究センター, 教授 (00199788)
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| Project Period (FY) |
1998 – 2000
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| Keywords | neutralizing antibody / anti-anti-idiotype / HIV / chemokine receptor |
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| Research Abstract |
The V3 region of an external envelope protein gp120 of HIV plays a crucial role in its chemokine receptor usage as a coreceptor. We have succeed in inducing an anti-idiotypic antibodies in rabbits by immunizing them with three monoclonal antibodies (mAbs) to V3 region. We titrated the anti-idiotype antibodies by an assay measuring the inhibitory activity in the binding of the biotinylated mAb to its epitope peptide. Anti-idiotype activity was most prominent in the combination of certain mAb and its anti-mAb anti-idiotype. However, cross-reactivity of anti-idiotypic antibody to the other mAb was observed to some extent. Anti-idiotype IgG was purified by immunoaffinity chromatography and the purity of anti-idiotype was more than 90%. We then analyzed the effect of these mAbs and anti-idiotypes in the interaction of chemokine and chemokine receptor (CCR5, CXCR4). We used 125I labeled RANTES, MIPα, and SDF1. However, no inhibitory activity of the chemokine binding to CCR5 transfected CEM cells (CEM/CCR5) was observed for these mAbs and their anti-idiotypes. We attempted to induce anti-anti-idiotype to induce neutralizing antibodies in mice. Although we immunized mice in the same titer of purified anti-idiotypes strung neutralizing activity was observed for the anti-anti-RC25 as compared to the other two. We have been trying to get an anti-idiotypic mAbs. However we have not succeeded to establish clone(s) that have activity comparable to the rabbit anti-idiotypic antiserum. Because high activity anti-idiotypic mAb is essential for further elucidating the utility of the anti-idiotype in vaccine development and immunotherapy we are continuing an effort to get such mAb producing cells.
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Research Products
(11 results)
