Analysis of cellular factors influencing the replication of human immunodeficiency virus type I in human macrophages derived from blood of different healthy donors

We analyzed parameters influencing HIV-1 infectibility of cells of the mono cyte/macrophage lineage (MO/MAC) isolated from different healthy donors. Th e proportion of in vitro-infected cells and replication kinetics in differe nt donor MAC ranged from 0.03 to 99% p24 antigen-positive MAC and from unde tectable RT activity up to 5 x 10(6) cpm/ml/90 min, respectively. As a quan titative measurement for HIV-1 susceptibility of donor MO/MAC, we determine d TCID50 values of defined virus stocks which varied up to 3000-fold depend ing on the donor MAC used for titration. As host factors which may influenc e the viral infection we determined the expression of Virus receptors CD4, CCR5, CXCR4, and CCR3 as well as the secretion of the natural ligands of CC R5, which altogether showed no correlation with HIV-1 infectibility of the cells. Moreover, other MO-derived secretory factors which might affect vira l infection of these cells could be excluded. Furthermore, expression of ma turation-related antigens CD14, CD16, HLA-DR, and MAX.1/CPM was determined. Analysis of the reverse transcription process revealed that restricted HIV -1 infection was reflected by highly reduced or even undetectable full-leng th HIV-1 DNA formation, although early and intermediate transcripts appeare d, suggesting that viral replication is blocked after entry at the level of early reverse transcription, (C) 2001 Academic Press.