How Proinflammatory Cytokines Block T Cell Death In Vivo

描述(由申请人提供)：我们的目标是确定促炎细胞因子和源自Toll样受体(TLR)刺激的因子如何影响体内特定的CD4T细胞免疫。表征这一过程将通过定义新的方法来控制体内T细胞反应，同时将毒性降至最低，从而促进佐剂的开发。这一目标是基于我们最近的工作，该工作揭示了TLR4适配器之间出人意料的分工。一条途径是T细胞效应器分化和归巢至实体器官，另一条途径负责克隆扩增和诱导细胞存活。因此，揭示TLR4途径产生的佐剂反应如何影响T细胞行为，将使疫苗专家能够推动对抗原特异性CD4T细胞的预期效果。通过确定每个适配器如何影响T细胞的机制，这一目标将通过三个具体目标实现。在目标1中，我们将通过检测一个先天的细胞因子轴来确定TRIF如何控制T细胞效应器的分化和归巢，我们假设这个轴增强了这两种细胞的行为。来自这一目标的数据将有助于设计疫苗配方，帮助推动效应器T细胞向身体特定部位的迁移。在目标2中，将捕获特定的、注定要存活的CD4T细胞的蛋白质组图，并将其与相同的编程死亡的特定T细胞进行对比。我们将使用一种创新的液体分离方法来实现这一目标，这将使我们能够挖掘蛋白质组来寻找这些种群之间的差异变化。AIM 3将利用我们的新发现，证明经历过Ag的CD4T细胞可以促进佐剂毒性。我们将定义这一过程，并开发新的方法来对抗毒性，同时促进强烈的佐剂反应。因此，这项建议的结果将提供一个“佐剂与毒性”的比率，可以用来衡量TLR激动剂的有效性和安全性，重要的是为设计辅助疗法提供了一种循证的方法。

DESCRIPTION (provided by applicant): Our goal is to define how pro-inflammatory cytokines and factors derived from toll-like receptor (TLR) stimulation impact specific CD4 T cell immunity in vivo. Characterizing this process will improve adjuvant development by defining new methods to control in vivo T cell responses while minimizing toxicity. This goal is based on our recent work which unveiled an unexpected division of labor between the TLR4 adaptors. One path signaled T cell effector differentiation and homing to solid organs, and the other path was responsible for clonal expansion and inducing cell survival. Therefore, uncovering how the adjuvant response emanating from the TLR4 pathway affects T cell behavior will allow vaccinologists to push desirable effects on Ag-specific CD4 T cells. This goal will be accomplished in 3 specific aims by determining the mechanism of how each adaptor impacts T cells. In Aim 1 we will determine how TRIF controls T cell effector differentiation and homing by examining an innate cytokine axis that we hypothesize potentiates both cell behaviors. Data from this aim will help design vaccine formulations that will aid in driving effector T cell migration to specific sites of the body. In Aim 2 a proteomic map of specific CD4 T cells destined to survive will be captured and contrasted to the same specific T cells programmed to die. We will accomplish this goal using an innovative liquid fractionation approach that will permit us to mine the proteome for differential changes between these populations. Aim 3 will capitalize on our novel finding demonstrating that Ag-experienced CD4 T cells can facilitate adjuvant toxicity. We will define this process and develop new approaches to counter toxicity while promoting strong adjuvant responses. Thus, the results from this proposal will provide an "adjuvant to toxicity" ratio that can be used to gauge the effectiveness and safety of TLR agonists, and importantly provide an evidence-based approach to designing adjuvant therapeutics.