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Characterization and epitope mapping of neutralizing monoclonal antibodies produced by immunization with oligomeric simian immunodeficiency virus envelope protein.
| Content Provider | Semantic Scholar |
|---|---|
| Author | Edinger, Aimee L. Ahuja, Ména Sung, T. Baxter, Kate Haggarty, Beth S. Hoxie, James A. |
| Copyright Year | 2000 |
| Abstract | In an attempt to generate broadly cross-reactive, neutralizing monoclonal antibodies (MAbs) to simian immunodeficiency virus (SIV), we compared two immunization protocols using different preparations of oligomeric SIV envelope (Env) glycoproteins. In the first protocol, mice were immunized with soluble gp140 (sgp140) from CP-MAC, a laboratory-adapted variant of SIVmacBK28. Hybridomas were screened by enzyme-linked immunosorbent assay, and a panel of 65 MAbs that recognized epitopes throughout the Env protein was generated. In general, these MAbs detected Env by Western blotting, were at least weakly positive in fluorescence-activated cell sorting (FACS) analysis of Env-expressing cells, and preferentially recognized monomeric Env protein. A subset of these antibodies directed toward the V1/V2 loop, the V3 loop, or nonlinear epitopes were capable of neutralizing CP-MAC, a closely related isolate (SIVmac1A11), and/or two more divergent strains (SIVsmDeltaB670 CL3 and SIVsm543-3E). In the second protocol, mice were immunized with unfixed CP-MAC-infected cells and MAbs were screened for the ability to inhibit cell-cell fusion. In contrast to MAbs generated against sgp140, the seven MAbs produced using this protocol did not react with Env by Western blotting and were strongly positive by FACS analysis, and several reacted preferentially with oligomeric Env. All seven MAbs potently neutralized SIVmac1A11, and several neutralized SIVsmDeltaB670 CL3 and/or SIVsm543-3E. MAbs that inhibited gp120 binding to CD4, CCR5, or both were identified in both groups. MAbs to the V3 loop and one MAb reactive with the V1/V2 loop interfered with CCR5 binding, indicating that these regions of Env play similar roles for SIV and human immunodeficiency virus. Remarkably, several of the MAbs generated against infected cells blocked CCR5 binding in a V3-independent manner, suggesting that they may recognize a region analogous to the conserved coreceptor binding site in gp120. Finally, all neutralizing MAbs blocked infection through the alternate coreceptor STRL33 much more efficiently than infection through CCR5, a finding that has important implications for SIV neutralization assays using CCR5-negative human T-cell lines. |
| File Format | PDF HTM / HTML |
| Alternate Webpage(s) | http://jvi.asm.org/content/74/17/7922.full.pdf |
| PubMed reference number | 10933700v1 |
| Volume Number | 74 |
| Issue Number | 17 |
| Journal | Journal of virology |
| Language | English |
| Access Restriction | Open |
| Subject Keyword | Cell Separation Cross Reactions Enzyme-Linked Immunosorbent Assay Epitope Mapping Epitopes Fluorescence Gene Products, env HIV Antibodies HIV Envelope Protein gp120 HIV-1 Hybridomas Immunologic Deficiency Syndromes Leukemia, B-Cell Ligand Binding Domain Monoclonal Antibodies Neutralization (Chemistry) Propionibacterium acnes Protocols documentation Simian immunodeficiency virus Subgroup V3 Loop Western Blotting fat-soluble vitamin polysaccharide-K virus envelope |
| Content Type | Text |
| Resource Type | Article |
