Modeling of HIF-1-alpha Regulation of B Cell Migration

项目摘要 对感染或疫苗接种的原发性人类免疫应答的表征通常依赖于测量 外周淋巴细胞，作为进入初级反应发生部位（例如引流淋巴）的通道 淋巴结（LN）由于解剖位置和伦理考虑而不容易接近。因此，大多数 的研究检查人B细胞活化和浆细胞分化在体外进行， 常氧组织培养条件。然而，B细胞在低氧环境中被激活、分化， 淋巴结的中心，并随后从一个解剖隔室迁移到另一个解剖隔室， 遇到不同的隔室氧水平，范围从淋巴结中的1-5%到淋巴结中的5-13%。 外周血在主动免疫反应期间，氧气水平对B细胞的影响，特别是 移民，未知。我们假设氧张力是一种以前未被认识到的B细胞调节因子， 开关、改变趋化因子受体信号传导和控制B细胞迁移。为了验证这一假设， 提出以下具体目标。目的1：定义和建模转录组和蛋白质组通路 负责B细胞迁移能力的O2依赖性功能变化。目标2：对重叠进行建模 转录组和蛋白质组途径负责钙调磷酸酶扰动的HIF-1α分子开关， 人B细胞。目的3：评估HIF-1α稳定化对小鼠疫苗效力的体内影响 免疫抑制模型拟议的工作旨在填补我们对以下问题的理解中的一个关键空白： 人B细胞应答，特别是不同氧水平对B细胞迁移的定量影响。

Project Summary Characterization of primary human immune responses to infection or vaccination often relies on surveying peripheral lymphocytes, as access to the sites where primary responses occur, such as the draining lymph nodes (LN), are not readily accessible due to anatomical location and ethical considerations. Thus, the majority of studies examining human B cell activation and plasma cell differentiation are performed in vitro under normoxic tissue culture conditions. However, B cells are activated, differentiate, within the low oxygen germinal center of the lymph node, and subsequently migrate from one anatomical compartment to another, where they encounter differing compartmental oxygen levels, ranging from 1-5% in the lymph node to 5-13% in the peripheral blood. The effects that oxygen levels have on B cells during an active immune response, in particular migration, is unknown. We hypothesize that oxygen tension is a previously unrecognized B cell regulatory switch, altering chemokine receptor signaling and controlling B cell migration. To test this hypothesis we propose the following specific Aims. Aim 1: Define and model the transcriptome and proteome pathways responsible for O2 dependent functional changes in B cell migratory capacity. Aim 2: Model the overlapping transcriptome and proteome pathways responsible for calcineurin perturbation of the HIF-1α molecular switch in human B cells. Aim 3: To assess the in vivo impact HIF-1α stabilization has on vaccine efficacy in a mouse model of immune suppression. The proposed work is intended to fill a critical gap in our understanding of human B cell responses, specifically the quantitative effects differing oxygen levels have on B cells migration.