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Session 49 Poster Session Vaccine Related Studies Session Time: 4:30-6:30 pm Room 4E-F

295-W. Contamination of Fetal Bovine Serum with Bovine Viral Diarrhea Virus: An Issue for Manufacturers of HIV-1 Vaccines L. Jagodzinski*1, J. Cooley1, S. Kelly1, T. VanCott1, and N. Michael2 1Henry M. Jackson Fndn. and 2Walter Reed Army Inst. Res. Program, Rockville, MD Background: Fetal bovine serum (FBS) is used for cultivation of chicken embryo cells in the production of HIV-1 MVA vaccine. Each lot of FBS is prepared by pooling several hundred serum bleeds from newborn calves. Bovine viral diarrhea virus (BVDV) is prevalent in the bovine population. Commercial suppliers perform biological testing for BVDV, but are not required to report the results unless requested by the purchaser. In addition, a lot may have no detectable biological activity, but may have significant levels of BVDV. To address potential BVDV contamination of HIV-1 MVA vaccine, a RT-PCR assay was developed and used to test twenty lots of FBS obtained from nine commercial suppliers. Method: A 2-step amplification procedure was developed as a semi-quantitative assessment of the BVDV viral RNA level. A 1-step RT-PCR procedure was used to amplify the 5 prime non-coding region of the BVDV viral RNA generating a PCR product of ~300 base pairs. PCR product was evaluated by gel electrophoresis and BVDV viral RNA levels were estimated by comparison to a known stock of BVDV. Serums having no visible amplification product or faint amounts of PCR product were subjected to an internal nested PCR amplification. Result: Of the 20 lots of FBS tested, 1 lot was considered to have no detectable BVDV viral RNA. 3 lots of FBS had levels of BVDV RNA detected by the nested approach (<10 infectious units/mL). 16 FBS lots had BVDV viral RNA levels corresponding to 103-106 infectious units/mL based upon amplification results observed for a BVDV viral stock. Conclusions: 95% of the FBS lots screened were contaminated with detectable amounts of BVDV viral RNA. Since BVDV can infect chicken embryo cells and thus be co-propagated with HIV-1 MVA, HIV-1 MVA vaccine preparations could be contaminated with BVDV. The injection of BVDV-contaminated HIV-1 MVA vaccine into immune compromised or healthy individuals could lead to immunological complications that could compromise HIV-1 vaccine trials.

©2002 9th Conference on Retroviruses and Opportunistic Infections