We seek to study virus entry and early mucosal spread after orally exposing infant rhesus monkeys (RMs) to an R5 SHIV in a model of breast milk transmission. This route of mother-to-child transmission (MTCT) continues to be problematic in developing countries. The risks of milk-borne MTCT have been linked to viral loads in lactoserum, among other parameters. It is not known, however, whether transmitted virions are covered by maternal antibodies (Abs), and if so, whether opsonization influences virion infectivity as well as the types and/or numbers of the first productively infected target cels. Since many nursing HIV+ women are seropositive, virions are likely opsonized. The possibility of Ab-mediated enhancement of HIV acquisition was raised recently. Willey et al.  showed that sera from individuals with recent HIV infection, who had not yet developed nAbs to autologous virus, rendered HIV infection in vitro significantly more virulent in the presence of complement - up to 350-fold. The effect was mediated by IgG and IgM. Using SHIV challenges in neonatal RMs, we have generated proof-of-concept that oral virus transmission is completely preventable by passive immunization with broadly neutralizing monoclonal antibodies (bnmAbs). We now seek to test the hypothesis that virions opsonized with non-neutralizing Abs (non-nAbs) will be associated with FcR-bearing or complement receptor (CR)-bearing cells after tonsilar virus application in RM infants. This in turn will be associated with a larger number of virus+ cells in the tonsils and adjacent oral mucosal tissues and lymph nodes. We also postulate that passive immunization with non-nAbs of infant RMs will increase their susceptibility to viremia after oral SHIV exposure.
The Specific Aims are to: 1. Examine the physical status of virions found in breast milk of HIV clade C-positive women and in milk of R5 clade C SHIV (termed SHIV-C)-infected RMs. We will test whether virions are Ab coated using virion- immune complex capture assays and whether such virions bind to FcR-and CR-bearing cells ex vivo. 2. Identify and enumerate the first virus target cells after exposing infant RMs via the tonsils to fluorescently labeled virus prepared either in standard culture medium or opsonized with non-nAbs. These studies will be conducted with single-cycle virions labeled either with green or red fluorescent proteins +/- opsonization with RM non-nAbs. 3. Test whether passive immunization of RM infants with non-neutralizing IgG isolated from R5 SHIV-C- challenged RMs with early-stage infection (before developing nAbs) will increase the infants' susceptibility to oral R5 SHIV-C challenge. We will use endpoint titration to determine the minimal infectious virus dose in orally challenged naive infants versus infants passively immunized with non-neutralizing IgG prior to virus challenge. The proposal is significant due to its focus on oral transmission and spread of opsonized virus, the most likely form to be involved in milk-borne HIV transmission given that most infected, lactating women are seropositive.
This project is significant because it seeks to analyze whether antibodies that arise early in the course of AIDS virus infection have the potential to increase viral spread in a primate model of oral transmission via breast milk~ significant enhancement of viral infection by such early antibodies has been suggested by a number of experiments in cultured cells. The project also seeks to analyze what types of antibodies cover HIV particles in breast milk of infected; seropositive women. Lastly; the project seeks to determine whether antibodies bound to virus particles alter the interaction of virus with cells in the oral mucosa inprimate model studies involving fluorescently labeled viruses.
|Sholukh, Anton M; Byrareddy, Siddappa N; Shanmuganathan, Vivekanandan et al. (2014) Passive immunization of macaques with polyclonal anti-SHIV IgG against a heterologous tier 2 SHIV: outcome depends on IgG dose. Retrovirology 11:8|
|Watkins, Jennifer D; Sholukh, Anton M; Mukhtar, Muhammad M et al. (2013) Anti-HIV IgA isotypes: differential virion capture and inhibition of transcytosis are linked to prevention of mucosal R5 SHIV transmission. AIDS 27:F13-20|