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

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来源获得了主要资金，因此可以在其他CRISP条目中表示。所列机构为中心机构，不一定是研究者机构。本文观察了轮状病毒感染后圈养恒河猴（Macacamulatta）外周血中病毒特异性CD 4+和/或CD 8 + T淋巴细胞的出现。这些细胞介导的免疫应答是在实验性或自然感染后从无症状动物的粪便标本中分离出轮状病毒后测量的。分离出的病毒是一种新的猿轮状病毒株，我们将其命名为TUCH（杜兰大学和辛辛那提儿童医院）。再刺激外周T淋巴细胞的灭活的双层或三层TUCH轮状病毒颗粒含有VP 6或VP 4和VP 7在其各自的表面上，导致细胞培养上清液中的白细胞介素-6（IL-6）和IL-12的量增加。CD 4+和CD 8 + T淋巴细胞对轮状病毒的回忆反应与细胞内IL-6和IFNg的积累有关。TUCH轮状病毒对淋巴细胞的抗原呈递通过单核细胞衍生的树突状细胞（HLA-DR+）的分化培养物介导。这是第一份证明非人灵长类动物对轮状病毒的细胞介导的免疫应答的报告。进一步探索恒河猴与人轮状病毒候选疫苗的疫苗试验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.