A Novel VSV/HIV Pseudotype Approach for the Study of HIV Microbicides Without Requirement for level 3 Biocontainment

Abstract

Studies of potential HIV mucosal microbicides are difficult to undertake due to the requirement for a suitable animal model and the use of biosafety level 3 containment, which are not always available to researchers. Here we show the use of a mouse model of vaginal and rectal transmission of an HIV chimeric virus that does not require level 3 biosafety containment, to test the ex vivo efficacy of soluble Gb3 analogs for the prevention of mucosal HIV infection. The model uses a pseudoenvelope-typed vesicular stomatitis virus (VSV)/HIV recombinant virus that can infect all murine cell types. We demonstrate that the envelope glycoproteins VSV-G of VSV and gp-120 of HIV both bind Gb3. We show that soluble Gb3 analogs inhibit in vitro infection of cervical and vaginal-derived cell lines by both intact HIV and the VSV/HIV recombinant virus. Soluble Gb3 analogs incorporated into gel or used alone and applied directly to the vaginal and rectal mucosal tissue of mice were able to resist viral infection as monitored by PCR and quantitative real-time PCR copy number of HIV cDNA extracted from mouse tissue. Only a trend towards significant efficacy for prevention of mucosal transmission through lower copy number in the treatment groups was evident from these studies; however, this finding warrants further evaluation. In addition, we illustrate a methodology to evaluate inflammatory responses in either vagina or rectum after administration of soluble microbicidal compounds. These studies provide a potential new ex vivo methodology suitable for animal facilities in general, to screen microbicide drug candidates, including drug candidates that target viral proteins, for efficacy and safety, in order to accelerate development and discovery of prophylactic and therapeutic agents for HIV/AIDS.