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

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 我们的合作者使用了老年和成年恒河猴(总共9只)，植入了遥测设备，每隔5分钟连续监测活动和温度，并实验感染野生型西尼罗河病毒(NY-99株)。这些设备在感染前7-14天植入，并在感染第一组和第二组后128天取出。在第二个队列中检测到了正常的昼夜节律周期，这一周期在第14天得到解决。没有一只动物在饲养员每天记录的两次行为中出现显著变化。 用定量聚合酶链式反应(QPCR)检测西尼罗河病毒基因组，发现虽然可以检测到初始水平，但在皮下感染1×10(7)pfu/只动物后，D100将病毒从血液中清除。此外，我们无法从从这些动物收集的任何材料中培养出传染性病毒，即使当qPCR表明存在mRNA(未显示)时也是如此。第二组，静脉注射。和S.C.每只动物感染1×10(9)pfu病毒在D10前被清除。 作为另一项措施，我们测试了猕猴的尿液，以评估病毒脱落的可能性。在接受1×10(7)pfu/只亚Q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D10开始，一直持续到D100，而在静脉注射之后。+S.C.感染1×10(9)pfu/只动物，尿液中未检出病毒。两组受试者都产生了瞬时的免疫球蛋白G反应。 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100天后没有感染西尼罗河病毒。年龄不影响临床疾病或病毒脱落。 对感染280天后因无关原因自发死亡的一只动物的组织学检查证实，没有西尼罗河病毒产生的损害，也没有免疫荧光检测到病毒。

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.