AEGiS-14IAC: Exposure of neutralization epitope on recombinant HIV-1 envelope glycoproteins expressed in BHK-21, HeLa or primary mouse cells.

14th International AIDS Conference


Barcelona, Spain - July 7-12, 2002

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Exposure of neutralization epitope on recombinant HIV-1 envelope glycoproteins expressed in BHK-21, HeLa or primary mouse cells.

Int Conf AIDS 2002 Jul 7-12; 14:(abstract no. A10009)

Rolland K, Brand D, Barin F
D3M, Virology Laboratory, Medicine School, Tours, France

BACKGROUND: The HIV-1 envelope glycoproteins are the primary target of neutralizing antibodies and may be an important component of an efficient HIV vaccine. Differences in the exposure of epitopes on the envelope surface were described between T-cell line adapted strains and primary isolates, and also from strain to strain. The objective of this work was to determine whether the exposure of various neutralizing epitopes was conserved when different cells are used for their production.

METHODS: HIV-1 envelope glycoproteins from 12 isolates were expressed using the Semliki Forest Virus expression system in BHK-21, HeLa and primary mouse embryo fibroblast cells. To assess the antigenic properties of each envelope, ELISA tests were performed using cell lysates and culture supernatants. Following the capture of the glycoproteins by an anti-gp120 antibody (D7324), the binding ability of the IgG1b12, 2G12 and 2F5 monoclonal antibodies and soluble CD4 to the envelope glycoproteins was determined.

RESULTS: Differences in the binding of monoclonal antibodies or/and soluble CD4 to envelope glycoproteins were observed in relation with the cell used for the protein expression for 7 of the 12 studied envelopes. The main differences concerned the binding of IgG1b12 (4 envelopes) and binding of soluble CD4 (4 envelopes).

CONCLUSION: These results support evidence that the cell line used for the protein expression can modify exposure of the neutralizing epitopes and the CD4 binding site on the HIV-1 envelope. These data must be considered when immunogenicity of HIV vaccine candidates based on recombinant envelope proteins is evaluated.

Keywords: AEGIS, HIV-1, Epitopes, Antigens, CD4, AIDS Vaccines, Semliki forest virus, Antibodies, Monoclonal, T-Lymphocytes, Enzyme-Linked Immunosorbent Assay, Recombinant Proteins, Cell Line, Glycoproteins, SK&F 106528, Mice, Animal, immunologyKWDaegis,hiv-1,epitopes,antigens,cd4,aidsvaccines,semlikiforestvirus,antibodies,monoclonal,t-lymphocytes,enzyme-linkedimmunosorbentassay,recombinantproteins,cellline,glycoproteins,sk&f106528,mice,animal,immunology

020707
A10009

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