SIV external envelope glycoprotein is the target of neutralizing antibodies.

Important note: Information in this article was accurate in 1991. The state of the art may have changed since the publication date.

SIV external envelope glycoprotein is the target of neutralizing antibodies.

Symp Nonhum Primate Models AIDS. 1990 Nov 28-30;8:47 (abstract no. 31). Unique Identifier : AIDSLINE PRIM8/900031
Collignon C; Kent K; Desrosiers R; DeWilde M; Bruck C; Thiriart C

Abstract: To investigate the antigenic and biological properties of SIV envelope glycoproteins, antibodies to SIV envelope antigen (molecular clone BK28) were obtained: i) a panel of monoclonal antibodies was isolated from a mouse immunized with SIV GP150 expressed in mammalian cells and ii) polyclonal antibodies were raised to three ENV subfragments expressed in E. coli. Monoclonal antibodies (mAbs) were characterized by competition EIA. 7 MAbs shown to react with nonoverlapping epitopes of SIV GP150 were chosen for further study: Western Blot, RIPA, cell surface reactivity (immunofluorescence on live cells) as well as reactivity to HIV-2 glycoproteins. Two epitopes were located on GP32, 5 on GP110. Whereas 6 out of 7 epitopes could be assigned to bacterial recombinant subfragments of GP150, 1 epitope appeared to be conformational and was not represented on any of the bacterial products. One mAb to GP32 and 1 mAb to GP110 showed cross reactivity to HIV-2 envelope. A 96 Well microtiter SIV neutralization test was developed; for this, CEMX 174 are used as indicator cells and SIVMM251 expanded on HUT 78 is used as viral inoculum. On day 4, the presence of viral antigen in the wells is monitored by a p57 capture EIA. Culture supernatant as well as ascitic fluid of the 7 selected mAbs were tested. Two mAbs displayed neutralizing activity. One was reactive in EIA and WB analysis with the NH2 terminal moiety of the SIV GP110 expressed in E. coli. The other one is unreactive to any E. coli recombinant fragments and appears to react with a conformational epitope of SIV GP110. Neutralizing activity was also detected in the polyclonal sera directed to one of the 3 bacterially expressed subfragments of SIV ENV. More precise mapping of neutralizing epitopes using synthetic peptides and determination of the cross specificity of neutralizing mAbs and sera are currently underway.

Keywords: Animal Antibodies, Monoclonal/*IMMUNOLOGY Antibodies, Viral/IMMUNOLOGY Antigens, Viral/*IMMUNOLOGY Binding, Competitive Cross Reactions Epitopes Escherichia coli/GENETICS Glycoproteins/GENETICS/*IMMUNOLOGY HIV-2/IMMUNOLOGY Macaca Neutralization Tests Protein Conformation Recombinant Proteins/IMMUNOLOGY SIV/*IMMUNOLOGY Viral Envelope Proteins/GENETICS/*IMMUNOLOGY ABSTRACT
910730
M9170963

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