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

Int Conf AIDS 1990 Jun 20-23; 6:319 (abstract no. 1022)
Shepp D, Ashraf A; North Shore University Hospital - Cornell University Medical College, Manhasset, NY, USA

OBJECTIVE: To determine if low-speed centrifugation of susceptible cell lines during viral adsorption can enhance isolation of HIV from stock virus preparations and from cell-free clinical sources.

METHODS: Duplicate cultures of 2.5x10(6) normal PHA-stimulated lymphoblasts (PHAB) or H9 cells were combined with 0.5 ml of serial 10-fold dilutions of HIV stocks or fresh serum and incubated for 15 minutes. One culture from each pair then was centrifuged at 700xg for 45 minutes (C) while the other remained in routine incubation (R). Cells were washed, placed in medium with 10% TCGF and refed twice weekly for 3-4 weeks. Growth of HIV was assessed by detection of p24 antigen in culture supernatants.

RESULTS: End-point titers of lab strain (HTLV-IIIB) and low passage clinical isolate ("MM") stocks were 1 and 2 log greater in H9 with C as compared to R. In PHAB, endpoint IIIB titers were 2 logs greater while "MM" titers were 1 log greater at day 7 of culture but equal at endpoint. C and R also were compared for recovery of HIV from patient serum: TABULAR DATA, SEE ABSTRACT VOLUME.

CONCLUSION: Low-speed centrifugation enhances isolation of laboratory and "wild" HIV strains in both H9 cells and PHAB. This method also facilitates recovery of HIV from sera of patients, especially those who are p24 antigen negative, have better clinical status and higher helper lymphocyte counts.

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