In FY91 DTS developed an urgent need to evaluate the safety of stored packed cells with regard to transmission of bacterial and viral agents. Concern about the risk of sepsis and death due to transfusion of yersinia- contaminated packed red blood cells (prbc's) prompted the Blood Advisory Committee to recommend reducing the storage time for packed cells. Available data suggests that deaths associated with yersinia contaminated units tend to increase with the time cells are stored at 2-6 C. In contrast, evidence suggests that the risk of transmission of HIV by transfusion of contaminated units increases with the blood storage time. The techniques developed under project 626 were used to experimentally determine the survival of cell-free HIV and HIV-infected cells in pbrc's stored at 2-6 C. Pbrc's from normal donors were prepared either in CPDA or ADSOL anticoagulant and maintained in satellite collection bags. Approximately 5 x 10o HIV-infected H9 cells and 6 x 10o cell-free virus particles (by EM) were added per ml pbrc's. Samples of inoculated and control prbc's were cryopreserved in 10% DMSO at various times over 21 days. Infectivity of samples was determined by 5 day co-culture with the indicator T-cell line containing the tat-sensitive HIV-LTR-CAT gene which expresses enzyme activity in response to infection. Results showed that infectious virus declined exponentially in both CPDA and ADSOL stored prbc's with estimated t 1/2 of 2.7 and 5.9 days, respectively. This work is being presented at the annual AABB meeting (Abstract #2145). Experiments are currently ongoing to determine if the anticoagulant used to preserve pbrc's has a direct effect on HIV survival in pbrc's stored at 2- 6oC.
