IMMUNE FUNCTION AND BIODEFENSE IN CHILDREN, ELDERLY, AND COMPROMISED POPULATIONS

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。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 研究中心，而研究中心不一定是研究者所在的机构。 我们在ONPRC的合作者使用了老年和成年恒河猴（共9只），这些恒河猴植入了遥测装置，以5分钟的间隔连续监测活动和温度，并实验性地用野生型西尼罗河病毒感染它们（菌株NY-99）这些器械植入7-第一组在感染前14天取出，第二组在感染后64天取出，第三组在感染后128天取出。在第二个队列中记录到正常的昼夜节律周期，并在第14天消退。 没有一只动物表现出明显的行为变化，每天记录两次，由他们的处理程序。 使用定量PCR（qPCR）检测西尼罗病毒基因组，发现尽管初始水平可检测到，但在皮下感染1 x 10（7）pfu/动物后，病毒在第100天从血液中清除。此外，我们不能从从这些动物收集的任何材料中培养感染性病毒，即使当qPCR指示mRNA的存在时（未显示）。在第二组中，在i. v.和s.c.用1 × 10（9）pfu/动物感染，病毒在第10天被清除。 作为额外的措施，我们测试了猕猴的尿液，以评估病毒脱落的可能性。 在给予1 x 10（7）pfu/动物的subQ感染的动物组中，我们使用尿液的qPCR数据显示病毒基因组从第10天开始持续到第100天，而在i. v.+ s.c.感染1 x 10（9）pfu/动物，尿液中从未检出病毒。两个队列均出现一过性IgG应答。 根据我们对它们的血液和尿液的分析，我们认为这些动物在感染后100天没有感染西尼罗河病毒。年龄不影响临床疾病或病毒脱落。 对一只在感染后280天因无关原因自发死亡的动物进行组织学检查，证实没有由西尼罗河病毒产生的病变，免疫荧光法也没有病毒。

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. Our collaborators at ONPRC have used aged and adult rhesus monkeys (9 total) that were implanted with telemetry devices which continuously monitored both activity and temperature at 5 minute intervals and experimentally infected them with wild type West Nile Virus (strain NY-99) These devices were implanted 7-14 days prior to infection and were removed 64 days post infection for the first cohort and 128 days post infection for the second cohort. Normal circadian cycles were recorded detected in the second cohort which resolved by day 14. None of the animals displayed significant alterations in behaviors recorded twice daily by their handlers. Quantitative PCR (qPCR) was used to detect West Nile viral genomes and found that although an initial level was detectable, the virus is cleared from the blood by d100 after subcutaneous infection of 1 x 10(7)pfu/animal. Moreover, we were not able to culture infectious virus from any of the materials collected from these animals, even when qPCR indicated the presence of mRNA (not shown). In the second group, following i.v. and s.c. infection with 1 x 10(9)pfu/animal the virus was cleared by d10 . As an additional measure, we tested urine from the macaques to evaluate the possibility of viral shedding. In the group of animals given sub Q infection of 1 x 10(7) pfu/animal, our qPCR data using urine revealed viral genomes starting at d10 which continued until d100 whereas after i.v. + s.c. infection of 1 x 10(9) pfu/animal, virus was never detected in the urine. Both cohorts developed transient IgG responses. Based on our analysis of both their blood and urine we believe the animals to be free from West Nile virus 100 days post infection. Age did not affect clinical disease or viral shedding. Histologic examination of tissues from one animal that died spontaneously for unrelated reasons 280 days after infection confirmed no lesions produced by WNV and no virus by immunofluorescence.