PRIMARY STIMULATION OF HUMAN CD8+ T LYMPHOCYTES

CD 8 + T淋巴细胞在人类免疫中对许多病毒性疾病和肿瘤的重要性已经得到充分证实。树突状细胞（Dendritic cells，DCs）是免疫应答的主要抗原提呈细胞（antigen presenting cell，APC）。事实上，目前正在广泛研究体内和离体靶向DC以用于肿瘤和病毒疫苗候选物的抗原呈递的方法。因此，更好地了解在初级刺激过程中人DC和CD 8 + T细胞之间的分子相互作用是非常需要的。 为此，我们选择研究同种异体DC对人CD 8 + T细胞的初级刺激。该系统具有许多优点，包括HLA类限制性刺激足够数量的CD 8 + T细胞，以允许检测和利用来自供体PBMC培养物的未成熟DC（iDC）和成熟DC（mDC）两者用作APC。目标1.将定性和定量评估同种异体APC，特别是iDC和mDC刺激初级CD 8 + T淋巴细胞应答的能力。将评价四种不同的标准：（a）T细胞标志物（CD 45 RA、CD 62 L）的上调（CD 25、CD 69、CD 44、CD 45 RO）和下调，（B）增殖，以及（c）细胞毒性应答和（d）TC 1（IFN-γ和IL-2）与TC 2（IL-4和IL-5）细胞因子产生谱的差异。目标2.通过上述四个标准测量，将剖析参与人CD 8 + T淋巴细胞活化和分化的同种异体iDC和mDC上或由其分泌的因子。 (a)共刺激分子，CD 80和CD 86，在同种异体DC上表达的CD 8 + T细胞的活化和分化的作用将被检查。(b)将评估同种异体DC产生的IL-12对CD 8 + T细胞的活化和分化的作用。(c)将确定Th 2/TC 2细胞因子IL-4和IL-10在CD 8 + T细胞的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.