Expression of Bcl-2 and intracellular p24 in SP CD4+ and immature thymocytes.
Cells were isolated as described in “Materials and methods,” infected with a primary isolate HIV-1B-LAIp, and maintained in culture in the presence of IL-7 (5 ng/mL), TNF-α (1 ng/mL), IL-1 (10 ng/mL), and IL-6 (10 ng/mL) for 11 days. (A) Bcl-2 expression was analyzed by Western blot on cell extracts from both control (C) and infected cells (HIV) in comparison with β-actin as a protein of unchanged expression. Quantification was determined as a ratio of Bcl-2/β-actin after densitometry analysis of the autoradiogram. Data are presented as the mean of independent experiments carried out with thymuses from 5 different donors. (B) Bcl-2 expression was analyzed by immunostaining and flow cytometry in uninfected (white area) and infected thymocytes (gray area). Dotted line indicates isotypic control. The arrow indicates the shift of the peak of Bcl-2–positive cells on infection. The experiment presented here is representative of independent experiments carried out with thymuses from 2 different donors. (C) Bcl-2 and intracellular p24 expression was analyzed by double immunostaining and flow cytometry in HIV-infected thymocytes. Percentages of single- and double-labeled cells are indicated in each quadrant. The experiment presented here is representative of independent experiments carried out with thymuses from 2 different donors.
