EVALUATION OF SAFETY OF MONOCLONAL ANTIBODIES AGAINST HIV ENVELOPE AND ITS CELLUL

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(1) Goals of project: - To study the activities of monoclonal antibodies (mAbs) raised against HIV-1 envelope (gp120), CD4, gp120/CD4 complexes, or the HIV-1 co-receptors, on HIV-1 fusion/infection of primary human cells. - To examine the effects of such mAbs on the normal biological functions of primary human cells. (2) Experimental approach: - We have collected a panel of murine mAbs specific for the HIV-1 gp 120 or for the CD4 molecules, that bind with increased avidity to the gp120/CD4 complex and may be involved in co-receptor recruitment. - These mAbs were tested for their effects on HIV-1 fusion/infection (both M-tropic and T-tropic strains), and for their activity in various assays designed to measure the formation of tri-molecular complexes between gp120,CD4, and the HIV-1 co-receptors. (3) Major Findings: - MAb CG10 was generated against soluble gp120(LAI)/CD4 complexes. It is gp120-specific but stricly complex-dependent. It was recently shown to interact with the region in gp120 that "faces" and possibly interact with the co-receptor molecules on target cells. In fusion/infectivity assays, we found that the CG10 mAb does not block but rather enhances viral fusion. Both T-tropic and M-tropic env-mediated fusion was enhanced, albeit at different optimal doses. These findings suggest that mAbs such as CG10 may have deleterious rather than protective effects in vivo by increasing cell-to-cell viral transfer. - CD4-specific mAbs (CG1,7,8) that bind with 10 fold higher avidity to preformed CD4/gp120 complexes than to CD4, were tested in multiple assays of HIV-1 co-receptor (CXCR4 or CCR5) recruitment. In three different assays these mAbs were found to stabilize the association of co-receptor, gp120, and CD4 in tri-molecular comlexes. Such antibodies may be used as tools to disect the steps leading to HIV-1 cell entry. In vivo, they may lead to an increase in viral-cell entry resulting in increased viral loads.