A NONHUMAN PRIMATE MODEL FOR TESTING OF IMMUNO GENE THERAPIES FOR AIDS

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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. Peptides derived from the heptad repeat 2 region (HR2) of HIV gp41 (C-peptides) effectively inhibit entry of HIV at the level of virus membrane fusion. A retroviral vector (M87o) expressing a membrane-anchored C-peptide was developed that also has strong antiviral activity in T cell lines and primary T lymphocytes (Egelhofer et al., J. Virol., 2004). M87o was shown to inhibit viral entry more than 10 000fold in single round infection assays. For testing in non-human primates, the M87o vector was adapted to either the SIVmac239 or the SHIV89.6 C-peptide sequence. The antiviral activity of these constructs against HIV, SIVmac and SHIV89.6 was analyzed in cell lines and in rhesus PBL. Surprisingly, all three C-peptide types inhibited human as well as the simian viruses. In particular, M87o was highly effective against SIV and SHIV89.6 and can therefore be tested without modifications non-human primate models for AIDS. Furthermore, these results indicate that the fusion process is strongly conserved between different lentiviruses. In addition, a protocol for large-scale retroviral transduction of T cells from rhesus macaques was developed. T cells were collected by lymphapheresis, stimulated with anti-CD3 and anti-CD28 immobilized on beads, transduced with GALVenv-pseudotyped M87o and expanded to high cell numbers. This non-human primate model will allow the optimization as well as safety and efficacy testing of immuno-gene therapy protocols for AIDS.