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

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。从HIV gp41的七肽重复2区（HR2）衍生的肽（c肽）在病毒膜融合水平上有效抑制HIV的进入。一种表达膜锚定c肽的逆转录病毒载体（m870）在T细胞系和原代T淋巴细胞中也具有很强的抗病毒活性（Egelhofer et al.， J. Virol.）。, 2004)。在单轮感染试验中，m870被证明能抑制病毒进入10000倍以上。在非人灵长类动物中，m870载体适用于SIVmac239或SHIV89.6 c肽序列。在细胞系和恒河猴PBL中分析了这些构建体对HIV、SIVmac和SHIV89.6的抗病毒活性。令人惊讶的是，所有三种c肽类型都能抑制人类和类人猿病毒。特别是，m870对SIV和SHIV89.6非常有效，因此无需修改即可在非人类灵长类动物艾滋病模型中进行测试。此外，这些结果表明融合过程在不同慢病毒之间是高度保守的。此外，还开发了一种大规模逆转录病毒转导恒河猴T细胞的方案。T细胞通过淋巴采集，用固定在珠上的抗cd3和抗cd28刺激，用galvenv伪型m870转导，扩增到高细胞数。这种非人类灵长类动物模型将允许艾滋病免疫基因治疗方案的优化以及安全性和有效性测试。

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.