PRIMARY STIMULATION OF HUMAN CD8+ T LYMPHOCYTES

CD8+T淋巴细胞在人类对多种病毒疾病和肿瘤的免疫中的重要性已经得到了充分的证实。树突状细胞(DC)是启动免疫反应的主要抗原提呈细胞(APC)。事实上，以树突状细胞为靶点的体内和体外肿瘤和病毒疫苗候选抗原呈递程序正在被广泛研究。因此，更好地了解人DC和CD8+T细胞在初始刺激过程中的分子相互作用是非常必要的。为此，我们选择研究同种异体DC对人CD8+T细胞的初步刺激作用。该系统具有许多优点，包括对足够数量的CD8+T细胞的HLA类限制性刺激，以允许检测和获得供者PBMC培养的未成熟DC(IDCs)和成熟DC(MDCs)作为APC。目的1.对同种异体APC，特别是IDCs和MDCs刺激CD8+T淋巴细胞反应的能力进行定性和定量的评估。将评估四种不同的标准：(A)上调(CD25、CD69、CD44、CD45RO)和下调T细胞标志物(CD45RA、CD62L)，(B)增殖和分化(C)细胞毒反应和(D)Tc1(干扰素-γ和IL-2)与TC2(IL-4和IL-5)细胞因子产生谱。目的2.根据上述四项标准，对人CD8+T淋巴细胞活化和分化过程中同种异体IDCs和MDCs的因子及其分泌情况进行分析。(A)研究表达在同种异体树突状细胞上的共刺激分子CD8O和CD86在CD8+T细胞的激活和分化中的作用。(B)将评估同种异体DC产生IL-12对CD8+T细胞活化和分化的作用。(C)将确定Th2/TC2细胞因子IL-4和IL-10在CD8+T细胞同种异体刺激中的作用。

The importance of CD8+ T lymphocytes in human immunity against many viral diseases and tumors has been well established. Dendritic cells (DCs) have emerged as the premier antigen presenting cell (APC) the initiation of immune responses. In fact, the targeting of DCs in vivo and in ex vivo procedures for antigen presentation of tumor and viral vaccine candidates is being extensively investigated. Thus, a better understanding of the molecular interactions between human DCs and CD8+ T cells during primary stimulation is highly desirable. TO this end, we have chosen to study the primary stimulation of human CD8+ T cells with allogeneic DCs. This system has a number of advantages including the HLA class restricted stimulation of sufficient numbers of CD8+ T cells to allow detection and the availability of both immature DCs (iDCs) and mature DCs (mDCs) from donor PBMC cultures for use as APCs. Aim 1. The ability of allogeneic APCs, in particular iDCs and mDCs, to stimulate primary CD8 + T lymphocyte responses will be both qualitatively and quantitatively assessed. Four different criteria will evaluated: (a) up-regulation (CD25, CD69, CD44, CD45RO) and down-regulation of T cell markers (CD45RA, CD62L), (b) proliferation, and the differentiation of (c) cytotoxic responses and (d) TC1 (IFN-gamma and IL-2) vs. TC2 (IL-4 and IL-5) cytokine production profiles. Aim 2. The factors on or secreted by allogeneic iDCs and mDCs involved in human CD8+ T lymphocyte activation and differentiation, as measured by the above four criteria, will be dissected. (a) The roles of the costimulatory molecules, CD8O and CD86, expressed on allogeneic DCs in the activation and differentiation of the CD8+ T cells will be examined. (b) The role of IL-12 production by allogeneic DCs on activation and differentiation of CD8+ T cells will be evaluated. (c) The effect of the Th2/TC2 cytokines, IL-4 and IL-10, in allogeneic stimulation of the CD8+ T cells will be determined.