Monocyte differentiation and HIV replication after prolonged culture of peripheral blood mononuclear cells from HIV-infected individuals

Primary cultures of peripheral blood mononuclear cells (PBMC) from 51 HIVhemophiliac patients (HIV+ PBMC) were set up, allowing undisturbed cellular interaction in the absence of any exogenous stimuli. The optimum time for p24 detection was between 12 and 25 days. Infective virus was recovered fro m the culture supernatants (HIV+ SN) and the amount of p24 released ranged from 25 to 5300 pg/ml. Cells of the monocyte/macrophage (M/M) lineage were the main source of HIV in the HIV+ SN, as judged by intracellular staining of permeabilized cells with anti-p24 (KC57 monoclonal antibody) and flow cy tometry analysis. M/M activation, differentiation, and proliferation occurr ed along the culture before the peak of in vitro HIV replication. Release o f HIV p24 was highest in patients with > 200 CD4(+) T lymphocytes/mm(3) who did not receive highly active antiretroviral therapy (HAART), but it was s till detectable in 60-90% of patients who had responded to 1-2 years of HAA RT, reducing their plasma viral load to undetectable levels. It is proposed that this simple experimental system can be used to assess ongoing HIV inf ection of M/M with the patient's own viral variants. (C) 2001 Academic Pres s.