EVALUATION OF ROTAVIRUS RNA LOAD IN NON-HUMAN PRIMATES BY REAL-TIME PCR

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。观察了轮状病毒感染后圈养幼年恒河猴外周血中病毒特异性CD4+和/或CD8+T细胞的出现情况。从无症状动物的粪便标本中分离出轮状病毒后，在实验感染或自然感染后检测这些细胞介导的免疫反应。分离到的病毒是一种新的猴轮状病毒株，我们将其命名为图赫(杜兰大学和辛辛那提儿童医院S)。用表面分别含有VP6或VP4和VP7的灭活双层或三层轮状病毒颗粒再次刺激外周T淋巴细胞，可使细胞培养上清液中IL-6和IL-12的含量增加。CD4+和CD8+T淋巴细胞对轮状病毒的召回反应与细胞内IL-6和IFNG的积聚有关。Tuch轮状病毒对淋巴细胞的抗原提呈是通过单核细胞来源的树突状细胞(HLADR+)的分化培养介导的。这是第一个证明非人类灵长类动物对轮状病毒的细胞介导免疫反应的报告。在人类轮状病毒候选疫苗的疫苗试验中进一步探索恒河猴是未来研究的主要目标。

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. Appearance of virus-specific CD4+ and/or CD8+ T lymphocytes in peripheral blood of captive juvenile rhesus macaques (Macaca mulatta) was observed following rotavirus infection. These cell-mediated immune responses were measured following the experimental or natural infection after rotavirus was isolated from stool specimens of asymptomatic animals. The virus isolated was a new strain of simian rotavirus that we named TUCH (Tulane University and Cincinnati Children¿s Hospital). Re-stimulation of peripheral T lymphocytes by inactivated double- or triple-layered TUCH rotavirus particles containing either VP6 or VP4 and VP7 on their respective surfaces, resulted in increased quantities of interleukin-6 (IL-6) and IL-12 in cell culture supernatants. Recall responses to rotavirus by CD4+ and CD8+ T lymphocytes were associated with accumulation of intracellular IL-6 and IFNg. Antigen presentation of TUCH rotavirus to lymphocytes was mediated via differentiated cultures of monocyte-derived dendritic (HLA-DR+) cells. This is the first report demonstrating cell-mediated immune responses to rotavirus in nonhuman primates. Further exploration of rhesus macaques in vaccine trials with human rotavirus vaccine candidates is the major objective of future studies.