This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator.
The specific aims remain essentially unchanged, as follows. 1. Do mucosal fluids of rhesus monkeys inactivate different SHIV strains in vitro and in vivo? Does inactivation correlate with levels of secretory leukocyte protease inhibitor (SLPI)? In collaboration with Dr. Diane Shugars, we plan to compare the biological and antiviral properties of SLPI from humans and rhesus monkeys. 2. What is the relative permeability of intact oral, vaginal, and rectal mucosa to R5 SHIV in adult macaques? Do neonatal monkeys differ in their susceptibility to oral R5 SHIV from that of adult macaques (this latter question will be addressed at no extra cost to this proposal)? 3. Is R5 SHIV preferentially transmitted through the oral route when the inoculum contains a 1:1 mixture of X4 and R5 SHIV strains? 4. Do mucosal fluids of monkeys dually infected with an R5- and an X4 SHV strain harbor predominantly R5 strains? During this year, we completed the rectal titration study (Aim 2). We titrated a new R5 SHIV in adult rhesus macaques through the rectal mucosa. This new virus, SHIV-1157ipd3N4, is using a R5-tropic env gene from an African isolate, which was transmitted from a mother to her child. Eight animals were enrolled sequentially in the rectal titration and we determined the AID50 (animal infectious dose) using the Vacman program (Spouge, 1992, PNAS 89:7581-5). For the mucoal inhibibition studies (Aim 1), we collected mucosal (saliva, vaginal, and rectal fluids) from a significant number of monkeys. We have measured concentration of SLPI in those samples and evaluated their ability to inhibit different strains of SHIV in-vitro. We are currently analyzing all the obtained data to calculate significant correlations. In parallel to our studies with the SHIV-1157ipd3N4, we also passaged a newly constructed SHIV, SHIV-2873Ni, generated by using the env gene of a primary HIV clade C isolated from a Zambian infant that displayed notably rapid progression to AIDS. The env gene was derived from an HIV clade C strain that may have been transmitted orally. This passaging step is necessary for molecular immunodeficiency virus clones to replicate efficaciously in rhesus macaques. Blood transfer continued in this manner involving a total of 5 animals, the usual number needed for virus adaptation. We plan to continue our in vivo experiments for oral and vaginal titrations of the SHIV R5 strain. We will start in vitro inhibition assays for human and monkey SLPI, with different SHIV strains.
Showing the most recent 10 out of 912 publications
