Productive human immunodeficiency virus infection levels correlate with AIDS-related manifestations in the patient.

Abstract

Mononuclear cells were obtained from 71 human immunodeficiency virus type 1 (HIV-1) seropositive subjects presenting at first visit either as asymptomatic or with minor symptoms and with CD4 lymphocytes >550 per mm3 (group A, 35 patients) or as patients with AIDS, AIDS-related illnesses, or CD4 lymphocytes <400 per mm3 (group B, 36

Infectious amplification of wild-type human immunodeficiency virus from patients' lymphocytes and modulation by reverse transcriptase inhibitors in vitro

“…In this system, the naturally infected human PBMC contain a diversified source of virus inocula in their normal environment. This natural unpassaged infectious material is mitogenically activated for infection and amplification in contiguous permissive lymphocytes with kinetics inherent to cell-to-cell transmission (7,11,13,14). The limiting dilution technique was used to specify the number of infectious cells brought into contact with amplifier lymphocytes and to measure their apparent reduction upon in vitro inhibition.…”

“…In preliminary studies, aliquots of the cell specimens contained between 500 and 20,000 infectious cells per 107 peripheral blood mononuclear cells (PBMC) as determined by limiting dilution analysis (see below). The isolation of these PBMC using Ficoll-Hypaque gradients, their cytofluorometric characterization, and their freezing in dimethyl sulfoxide culture medium, storage in liquid nitrogen, thawing, washing, and counting at the time of culture were performed as described previously (13). Viability after the cells were thawed was 295% (13 Frequencies of infectious cells and virus scores.…”

“…The isolation of these PBMC using Ficoll-Hypaque gradients, their cytofluorometric characterization, and their freezing in dimethyl sulfoxide culture medium, storage in liquid nitrogen, thawing, washing, and counting at the time of culture were performed as described previously (13). Viability after the cells were thawed was 295% (13 Frequencies of infectious cells and virus scores. The method used for determining virus scores has been described previously (13).…”

“…Viability after the cells were thawed was 295% (13 Frequencies of infectious cells and virus scores. The method used for determining virus scores has been described previously (13). Briefly, the routine titration of infectious cells of patients uses conservative limiting dilutions with three dilutions per cell specimen, each of them fivefold apart, and six replicates for each cell concentration tested.…”

“…Briefly, the routine titration of infectious cells of patients uses conservative limiting dilutions with three dilutions per cell specimen, each of them fivefold apart, and six replicates for each cell concentration tested. To maximize the mathematical use of such limited dilutions, the viruspositive replicates from two contiguous dilutions were combined to yield virus scores, i.e., percentages of virus-positive wells from 6 + 6 proximate cell dilutions (13). Plotting virus scores for the 61 reference specimens against the frequencies of infectious cells provided a series of linear curves which allowed a direct readout of infectious-cell frequencies from a given virus score.…”