Recovery of HIV-related retroviruses from Italian patients with AIDS or AIDS-related complex and from asymptomatic at-risk individuals

Human retroviruses have been detected in supernatants of cultures of Ficoll-enriched lymphocytes from peripheral blood, lymph nodes and bone marrow of (a) 32 out of 42 patients with Acquired Immunodeficiency Syndrome (AIDS), (b) 34 out of 64 patients with AIDS-related Complex (ARC), (c) 9 out of 18 asymptomatic children born from Human Immunodeficiency Virus (HIV) seropositive mothers, and (d) 9 out of 28 asymptomatic drug abusers or hemophiliacs. Virus detection was monitored by assaying culture supernatants for the presence of Mg⁺⁺-dependent reverse transcriptase (R.T.) activity. A number of these virus-positive sups were passaged repeatedly in cultures of phytohemagglutinin-stimulated and Interleukin-2 (IL-2) treated fresh lymphocytes from healthy blood donors. Occasionally, multiple samples were obtained at varying time intervals from the same patient and consistently yielded detectable retroviral activity. Several isolates were characterized as closely related if not identical to HIV, HTLV-III(B) strain, since cells from either patients' own lymphocyte cultures or subcultures infected with passaged virus were stained in an indirect immunofluorescent assay with both patients sera and monoclonal antibody against p24 antigen of the HTLV-III(B) strain. Representative isolates, grown on fresh lymphocytes of healthy donors and metabolically labelled with ³⁵S-cysteine, were also analyzed in a radioimmunoprecipitation assay (RIPA) against patients' sera to define their antigenic pattern, which was widely superimposable to that obtained with HTLV-III(B)-infected H9 cells.DNA from lymphocytes infected with 2 representative isolated were Southern-blotted and probed with an insert from a plasmid containing the entire genome of the HTLV-III(B) strain. The hybridization patterns were comparable with those obtained with DNA from H9-infected cells.