Increased expression of CD80 and CD86 in in vitro-infected CD3(+) cells producing cytoplasmic HIV type 1 p24

Determining the effects of HIV infection on the expression of cell surface molecules has been limited by an inability to differentiate between product ively infected cells and those without productive infection. We inoculated human peripheral blood mononuclear cells from healthy, human immunodeficien cy virus type 1 (HIV) antibody-negative donors with HIV; noninoculated cell s were also examined. Using multiparameter flow cytometry, we differentiate d cells actively producing HIV cytoplasmic p24 antigen during acute, in vit ro HIV infection from those not producing detectable cytoplasmic p24. For b oth resting and PHA-stimulated cells inoculated with HIV (R/H and P/H), a h igher proportion of p24(+) cells expressed CD25, compared with p24(-) cells (p = 0.031 and p = 0.008, respectively), consistent with either increased viral replication in stimulated cells or increased stimulation secondary to productive HIV infection. Findings were similar for the expression of CD38 , HLADR, and CD28. A striking proportion of p24(+) cells expressed CD80 or CD86, antigens not usually expressed by CD3(+) lymphocytes. The increased e xpression appeared to be independent of stimulation status in that it occur red in both the R/H and P/H treatment groups but not in resting or PHA-stim ulated uninfected cells. CD28 expression was generally comparable between C D3(+) cells that did and did not express CD80 or CD86. Multiparameter flow cytometry, in association with improved techniques for cell permeabilizatio n and cytoplasmic fluorescent staining, should prove useful in examining th e effects of productive HIV infection on surface and cytoplasmic cellular m olecules. Using this approach, we found an association between productive i nfection and increased expression of CD80 and CD86. This association has im plications for HIV disease pathogenesis and, potentially, HIV therapy.