IMMUNE FUNCTION AND BIODEFENSE IN CHILDREN, ELDERLY, AND THE IMMUNOCOMPROMISED

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。子项目及 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。 我们正在使用西尼罗河病毒感染老年恒河猴作为老年人免疫功能的模型。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）这些器械在感染前7-14天植入，第一组在感染后64天取出，第二组在感染后128天取出。记录了正常的昼夜节律周期，尽管在一只动物（20437）中检测到发热，但其发生了直接归因于自然流产的并发症。 感染的其他指征是在第二个队列中检测到的轻微单核细胞增多症（在CBC分析期间测量），其在第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（如果可能，通过平底锅捕获或无菌膀胱穿刺术收集）的qPCR数据显示病毒基因组从d10开始持续到d100，而i. v.+ s.c.感染1 x 10（9）pfu/动物，尿液中从未检出病毒。如前所述，任何材料（包括检测到病毒RNA的尿液）的共培养均未产生任何感染性病毒迹象。 来自两个队列的动物成功地产生了IgG应答，其已经达到峰值并且现在正在衰减。 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年没有感染西尼罗河病毒。年龄不影响临床疾病或病毒脱落。

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. We are using West Nile virus infection of aged rhesus monkeys as a model for immune function in the elderly 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 and although a fever was detected in one animal (20437) she was experiencing complications directly attributable to a spontaneous abortion. The other indication of infection was a slight monocytosis (measured during CBC analysis) 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 (collected via pan catch or sterile cystocentesis if possible) 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 . As mentioned, co-culture of any of the materials, including urine where viral RNA was detected, did not produce any indication of infectious virus. The animals from both cohorts successfully developed IgG responses which have peaked and are now decaying. Based on our analysis of both their blood and urine we believe the animals to be free from West Nile virus 100 post infection. Age did not affect clinical disease or viral shedding.