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156 High Levels of Nonfunctional Proviral DNA in DNA/MVA Vaccinated Animals with Controlled Viral Infections
Y. Tang*, R. R. Amara, F. Villinger, S. Staprans, N. Kozyr, B. Grimm, J. Smith, S. Lydy, and H. L. Robinson
Emory Univ., Atlanta, GA, USA
Background: Proviral DNA is a useful maker for exploring viral reservoirs. Recently, we demonstrated that DNA/MVA-vaccinated animals were infected but rapidly controlled a SHIV-89.6P challenge to near or below the level of detection of plasma viral RNA by RT-PCR.
Methods: A taqman real-time PCR assay was developed to quantify SHIV-89.6P proviral DNA. The lower limit for detection of the assay is 5(10 copies of proviral DNA per microgram of DNA. Proviral DNA encoding replication-competent virus was tested for by cocultivation of PBMC from vaccinated animals with mitogen-stimulated PBMC from uninfected animals.
Results: At 7(8 months post challenge, vaccinated animals that had controlled viral loads to <300 copies of viral RNA per milliliter of plasma had proviral loads of over 100 copies per microgram of PBMC DNA. When normalized for CD4 cells, the loads represented >1,000 copies of provirus per 106 CD4 cells. These same animals were negative in cocultivation assays using 106 PBMC.
Conclusions: DNA/MVA vaccinated animals controlled viral loads more effectively than proviral loads. However, most of the proviral DNA was not replication competent.
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