Optimal conditions for recovery of the human immunodeficiency virus from peripheral blood mononuclear cells

Abstract

Optimal conditions for demonstrating the presence of infectious human immunodeficiency virus in peripheral blood mononuclear cells (PMCs) from seropositive individuals involved cocultivation of infected cells with phytohemagglutinin-stimulated PMCs from seronegative donors in the presence of 2 μg of Polybrene per ml. The size of the culture vessel also influenced the results; smaller numbers of infected cells were detected under conditions of increased cell density. In addition, an increased normal donor/patient PMC ratio was helpful. The cocultivation approach permitted identification of human immunodeficiency virus in over 90% of seropositive individuals with different clinical conditions. Moreover, reconstruction experiments indicated that this method allows detection of one productively infected CD4+ cell in a population of over 106 PMCs.