Detection of viral surface antigens on HIV‐2ben infected human tumor cell lines by flow cytometry
Dorothea Böhm, Sigrid Nick, Gérald Voss, Gerhard Hunsmann
Abstract
Dorothea Böhm, Sigrid Nick, Gérald Voss, Gerhard Hunsmann
Abstract
The human monocytic cell line U-937 clone 2 and two T-cell lines CEM and MOLT-4 clone 8 were infected with HIV-2ben, a recent isolate of HIV-2. Infection and subsequent antigen expression on the cell surface was monitored by flow cytometry using a rabbit-anti-serum against tween-ether-treated HIV-2ben and a fluorescein-isothiocyanate-conjugated IgG against rabbit-IgG. The sensitivity of the three cell lines to infection with HIV-2ben correlated with the percentages of CD4-expressing cells but not with the levels of CD4-expression on the cell. The appearance of viral surface antigens preceded the formation of syncytia and correlated closely with the infecting virus dose. After 1-2 weeks in culture, 20-85% of the cells of each line expressed viral surface antigens. The variation depended on the cell type and cell culture conditions. The MOLT-4 clone 8 and the U-937 clone 2 cells died around 10 or 20 days, respectively, after HIV-2ben infection. Only HIV-2ben infected CEM cells grew permanently. Flow cytometry was an appropriate method to monitor the expression of viral proteins on the cell surface of HIV-infected cell lines. Flow cytometry proved to be more sensitive than determination of RT activity in supernatants of HIV-infected cells and more precise than light microscopy examinations.
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The human monocytic cell line U-937 clone 2 and two T-cell lines CEM and MOLT-4 clone 8 were infected with HIV-2ben, a recent isolate of HIV-2. Infection and subsequent antigen expression on the cell surface was monitored by flow cytometry using a rabbit-anti-serum against tween-ether-treated HIV-2ben and a fluorescein-isothiocyanate-conjugated IgG against rabbit-IgG. The sensitivity of the three cell lines to infection with HIV-2ben correlated with the percentages of CD4-expressing cells but not with the levels of CD4-expression on the cell. The appearance of viral surface antigens preceded the formation of syncytia and correlated closely with the infecting virus dose. After 1-2 weeks in culture, 20-85% of the cells of each line expressed viral surface antigens. The variation depended on the cell type and cell culture conditions. The MOLT-4 clone 8 and the U-937 clone 2 cells died around 10 or 20 days, respectively, after HIV-2ben infection. Only HIV-2ben infected CEM cells grew permanently. Flow cytometry was an appropriate method to monitor the expression of viral proteins on the cell surface of HIV-infected cell lines. Flow cytometry proved to be more sensitive than determination of RT activity in supernatants of HIV-infected cells and more precise than light microscopy examinations.
Key concepts: Flow cytometry, clone (Java method), Cell culture, Antigen, Biology, Molecular biology, Syncytium, Cell