AEGiS-06IAC: Specific binding of vaccinia recombinant-derived HIV-1 gp 160 to CD4(+) cell lines.

6th International AIDS Conference

San Francisco, California, USA — June 20-23, 1990

Specific binding of vaccinia recombinant-derived HIV-1 gp 160 to CD4(+) cell lines.

Int Conf AIDS 1990 Jun 20-23; 6:329 (abstract no. 1060)
Kistner O, Barrett N, Mitterer A, Dorner F; Immuno AG, Vienna, Austria

OBJECTIVE: We have developed a method for large-scale production and purification of gp 160 from recombinant vaccinia virus-infected Vero cells. This protein has been shown to be able to elicit T-cell proliferative responses and cross-reactive neutralizing antibodies in animals. In this study we have analyzed the ability of the purified recombinant-derived protein to bind to the CD4 receptor.

METHODS: Binding of gp 160 to the CD4 receptor was determined by incubating the soluble protein with CD4+ and CD4- cells. After repeated washing, the cells were lysed and subjected to electrophoresis and Western blot analysis using human HIV-1 antiserum.

RESULTS: We have demonstrated that: 1) recombinant gp 160 binds to different CD4+ cell lines such as CEM, MT4 and H9, but not to CD4- cells such as Vero; 2) denatured recombinant gp 160 does not bind to CD4+ cells; 3) deglycosylated recombinant gp 160 does not bind to CD4+ cells; and 4) antiserum against gp 160 inhibits the binding of recombinant gp 160 to CD4+ cells.

CONCLUSION: The large-scale production and purification of vaccinia recombinant-derived gp 160 leads to a native and biologically active protein, which binds specifically to the CD4 receptor.

Keywords: AEGIS, Antigens, CD4, HIV-1, Cell Line, Sialoglycoproteins, Vaccinia virus, Blotting, Western, GP 160, Human, Animal, ICA6

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