Vanessa's Presentation

Repeated intravaginal inoculation with cell-associated virus results in persistent non-human primates

Background
• Young women most vulnerable to AIDS pandemic
• Worldwide heterosexual route is most prevalent
• Compromised tight epithelial cells
Objectives
• Develop a non-hman model of intravaginal HIV transmission with cel-associated virus
• Model will possibly help with testing of vaccines
Test Subjects
• Mature, female cynomologus macaque
o Experimental:
 Inspected for pre-existing inflammation
 Benzalkonium chloride
 Inoculums applied to ulcers
o Control:
 No inspection of detergent
 Inoculums deposited at vaginal entry
• Inoculums
o Peripheral blood mononuclear cells
o Incubated with SIVmac239
o Incubated 7-9 days with IL-2
o SIV-infected PBMCs were cocultured and serially split
o Dead cells and cell-free virus removed
• Measuring proviral DNA (see figure)
o Isolate all genomic DNA
o Perform PCR using primers specific to viral gag gene
o Parallel amplification of e-globin
o Used to determine proviral DNA copies
o Inoculums proviral DNA: 104-105 copies/million cells
• Viral challenge
o Viremia does not cause IL-6
o Low level of cells can cause viremia

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