DNA AND PROTEIN IMMUNOGENS FOR SIV/HIV VACCINES

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。在恒河猴中使用SHIV-89.6 DNA和MVA免疫原和SHIV-89.6P攻击，对GM-CSF作为DNA/MVA疫苗佐剂的能力进行了研究。这些研究表明，GM-CSF DNA与SHIV-89.6 DNA共同给药使峰值和设定点病毒载量降低了10倍，超过了在没有对T细胞进行GM-CSF研究的情况下接受疫苗DNA的动物中发现的病毒载量。由含GM-CSF佐剂的和无佐剂的疫苗引起的Ab应答显示，GM-CSF影响了Ab和T细胞应答。这些影响并不表现在反应的高度差异上，而是表现在反应的质量上。GM-CSF佐剂增加了引发的抗Env Ab的亲和力，扩大了应答的中和活性。中和活性的扩大延伸到相对容易中和的分离株，但不包括HIV-1的偶发株。当分析粘膜和全身反应时，发现激发后T细胞反应的主要差异。这些分析显示，GM-CSF佐剂化的动物在肠粘膜中具有比非佐剂化的动物更高的抗病毒CD 8 T细胞频率。这些频率差异持续至攻毒后8周。 还对2只接种疫苗和攻毒猴的T细胞应答及其与病毒序列突变的关系进行了研究，这些猴在控制4年后失去了对攻毒病毒的控制。失去病毒控制的两只猴子在响应CD 8和CD 4 T细胞的广度和频率上都经历了戏剧性的扩张。最初，这些抗病毒T细胞表达IFN-β，其中约20%共同产生IL-2。随着时间的推移，IL-2共同生产细胞，然后IFN-？产生细胞丢失，动物患上了艾滋病。

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. Studies were conducted on the ability of GM-CSF to serve as an adjuvant for a DNA/MVA vaccine using SHIV-89.6 DNA and MVA immunogens and a SHIV-89.6P challenge in rhesus macaques. The studies demonstrated that GM-CSF DNA co-administered with the SHIV-89.6 DNA decreased peak and set point viral loads by 10-fold over those found in animals that received the vaccine DNA in the absence of the GM-CSF studies on the T cell. Ab responses elicited by the GM-CSF-adjuvanted and non-adjuvanted vaccines revealed that the GM-CSF had affected both Ab and T cell responses. These effects were not manifest in differences in the heights of responses but rather in the quality of responses. The GM-CSF adjuvant increased the affinity of the elicited anti-Env Ab broadening the neutralizing activity of the response. This broadening of neutralizing activity extended to isolates that are relatively easy to neutralize, but not to incident strains of HIV-1. The major difference in T cell responses was found post challenge when mucosal as well as systemic responses were analyzed. These analyses revealed that the GM-CSF-adjuvanted animals had higher frequencies of anti-viral CD8 T cells in the gut mucosa than the non-adjuvanted animals. These difference in frequencies persisted for up to 8 weeks post challenge. Studies were also conducted on the T cell responses and their relationship to mutations in viral sequences in two vaccinated and challenged monkeys that lost control of the challenge virus after 4 years of control. Both of the monkeys that lost viral control underwent dramatic expansions in the breadth and frequency of responding CD8 and CD4 T cells. Initially, these anti-viral T cells expressed IFN-?, with about 20% co-producing IL-2. Over time IL-2 co-producing cells and then IFN-? producing cells were lost and the animals developed AIDS.