T cell responses to H1N1 and a longitudinal study of seasonal flu vaccination

在这里，我们建议描述人类志愿者对甲型H1N1v大流行疫苗接种或感染后的CD4[+]、CD8[+]和γ-Delta T细胞的反应。我们将使用两种系统生物学方法以及结合单细胞细胞因子分析结合多肽-MHC四聚体分析各种抗原特异性反应，在单细胞细胞因子分析中，我们可以筛选数百个不同的表位和数十个细胞因子。在目标1中，我们将询问特定的表位或细胞因子表达模式是否与强大的疫苗反应相关，如通过微量中和试验和/或颗粒酶B-T细胞试验测量的，或者在感染个体的情况下，疾病严重程度。通过这两项双胞胎研究，我们将询问显性表位选择与遗传身份的关系有多密切，以及这种情况是否会随着年龄的增长和接触传染病而改变。这将是对基因组学核心完成的完整的人类白细胞抗原基因分型所获得的结果的补充，因为我们预计人类白细胞抗原单倍型将对表位的选择和显性产生重大影响。在目标2中，我们将表征γ-增量T细胞中的激活标记和细胞因子的表达，最近的证据表明，它通过早期产生IL-17来影响抗体反应，并帮助塑造CD4[+]和CD8[+]T细胞和其他免疫系统成分的反应。在目标3中，我们将继续我们现在已有三年的关于年轻人(20-30岁)季节性流感疫苗接种的纵向研究。与年龄较大的人群(60-96岁)相比，在人类免疫监测核心和生物统计学核心的帮助下，采取了系统生物学的方法，其中我们将基因表达、细胞因子刺激和血清细胞因子与免疫衰老等参数相关联，以揭示许多老年人正常免疫功能失败背后的新标记和机制。我们已经发现了一些免疫缺陷的新标记物，包括B细胞中AID表达的减少和许多老年人对细胞因子的反应不足。这一机制的支持将使我们能够广泛地观察免疫衰老的年度变化和早期标志。最后，将采用同样的系统方法来比较H1N1v疫苗接种对实际感染的反应，这应该有助于我们了解两者之间的相似和不同之处，并有助于设计更具保护性的疫苗。

Here we propose to characterize the responses of CD4[+], CD8[+] and gamma-delta T cells to the pandemic influenza H1N1v vaccination or infection in human volunteers. We will use both systems biology approaches as well as analyzing a variety of antigen specific responses with combinatorial peptide-MHC tetramers combined with single cell cytokine analysis, in which we can screen for hundreds of different epitopes and dozens of cytokines. In Aim 1. we will ask whether particular epitope or cytokine expression patterns correlate with a robust vaccine response, as measured by a microneutralization assay, and/or a granzyme B T cell assay, or in the case of infected individuals, disease severity. With the twin studies we will ask how closely does dominant epitope choice follow genetic identity and does that change with increasing age and exposure to infectious diseases. This will be complementary to the results obtained with the complete HLA genotyping being done by the Genomics Core, as we expect that HLA haplotypes will have a major influence on epitope selection and dominance. In Aim 2. we will characterize activation markers and cytokine expression in gamma-delta T cells, which recent evidence suggests influence antibody responses and help shape the responses of CD4[+] and CD8[+] T cells and other immune system components through the early production of IL-17. In Aim 3 we will continue our now three year old longitudinal study of seasonal flu vaccination in young adults (20-30 yrs.) versus older cohorts (60-96 yrs), taking a systems biology approach with the help ofthe Human Immune Monitoring Core and the Biostatistics Core in which we correlate gene expression, cytokine stimulation and serum cytokines with parameters such as immune senescence to uncover new markers and mechanisms behind the failure of proper immune function in many older people. We have already identified a number of new markers of immune deficiency, including a decrease in AID expression in B cells and deficient responses to cytokines in many older people using phosphoflow analysis. Support by this mechanism will allow us to look extensively at year-on-year variations and early markers of immune senescence. Lastly, this same systems approach will be taken to compare H1N1v vaccination responses to actual infections, which should help us to understand the similarities and differences between the two and aid in the design of vaccines that are more protective.