6th International AIDS Conference San Francisco, California, USA — June 20-23, 1990

Int Conf AIDS 1990 Jun 20-23; 6:320 (abstract no. 1024)
Cuneo P, Sigari G, Losi E, Zemignan M, Arenare L, Bonafede L; Inst. Hyg. Prev Med University Genoa, Italy

OBJECTIVE: To evaluate the result of different techniques for HIV isolation from blood (whole blood, PBMC and plasma). Our aim is both monitoring clinical evolution in seropositive patients and the finding out the most useful method for HIV recovery in subjects with indetermined serological pattern or seronegative ones at risk.

METHODS: The following procedures were used in HIV isolation: 1) Patients' fresh PBMC (1x10) were stimulated with phytohemagglutinin (PHA) and cultured as in the procedure described by Gallo et al. 2) Patients' PBMC were co-cultured with normal donor PBMC which had been stimulated with PHA. Additional normal donors' PBMC stimulated with PH were added every 4-7 days to enhance the virus replication. 3) Subjects' whole blood (500 ul) was added to normal donors' PBMC, as in the previous procedure. 4) Patients' filtered plasma (2 ml) was added to PBMC culture as in the previous procedure. HIV replication in the cultures has been evaluated by HIV p24 antigen titration. Samples of cultured cells have been processed to be examined by transmission electron microscope. Laboratory and clinical data were available for all the patients.

RESULTS: Out of 115 anti-HIV positive subjects, the co-culture of PBMC, carried out according to the 2nd procedure, furnished the highest percentage of HIV isolation (85%). Even whole-blood culture gave satisfactory results in 15 samples studied in comparison to the former technique. Virus isolation was obtained from plasma of 35% out of 40 subjects investigated. It resulted more frequently in AIDS patients as compared to asymptomatic ones. The follow up of said asymptomatic subjects pointed out that plasma viremia should be considered negative from the prognostic point of view. Plasma viremia was often met with before the HIV Ag appearance. No HIV recovery has been obtained from 45 blood-donors with anti-HIV indeterminate serologic pattern. TEM examination of culture-cells allowed to verify the presence of virus particles in positive cases. Furthermore EM evidenced the presence of other viruses, especially HHV6.

CONCLUSIONS: HIV recovery from blood and plasma in seropositive subjects proves to be useful to classify the clinical stage of infection. Improvement has been obtained in isolation techniques. Plasma viremia shows a sensible prognostic marker.

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