SALIVARY IGA RESPONSES--REGULATION BY T CELLS

描述(改编自申请者的摘要)：这是竞争 更新RO1拨款，检查黏膜免疫反应，即 负责免疫球蛋白A B细胞反应的诱导和调节。这个 工作重点是小鼠的口腔/鼻腔(例如，颌下腺， SMG)。以往的研究表明，Th2细胞来源的IL-5和IL-6是 诱导SIgA阳性B细胞分化的特殊重要性 转化为产生IgA的浆细胞，以及这些细胞因子的高水平mRNA 以及干扰素-伽马，由SMGα-beta的研究人员报告 (AB)TCRCD4阳性T细胞。当前的应用程序建议检查 Gd T细胞在粘膜免疫反应中的作用。这 方向是基于这样一个事实，即已知gdT细胞定位于 它们约占IEL的50%的粘膜部位 肠道，研究人员发现大约20%的 SMG内的T细胞为Gd T细胞。调查员通过以下方式表明 ELISPOT分析表明SMG中主要的Ig产生细胞为IgA 提示SMG与肠道LP一样，是一种IgA效应组织。这个 SMG gd T细胞包括IL-4和IL-5产生细胞，但不包括IL-4 制片人。此外，调查人员在最近完成的调查中发现 Gd缺陷(基因敲除)小鼠的粘膜免疫球蛋白A反应受损。 除了粘膜免疫系统在产生 SIgA，粘膜诱导耐受是口腔中的一种重要现象 注射抗原可产生黏膜IgA反应，但全身性 反应迟钝。研究人员已经表明，粘膜中的gdT细胞来自 口服耐受小鼠的肠道上皮细胞可被清除 过继转移gdT细胞时抗原特异性无反应性。 其他人的研究表明，gdT细胞可以下调IgE 口服抗原后的反应。调查员提议 GdT细胞在粘膜IgA和E中可能起重要调节作用 粘膜部位通过细胞因子的产生和通过 与ab T细胞的相互作用。在第一个目标中，SMG单个核细胞将 用ELISPOT法检测Gd缺乏症患者的IgA产生细胞 野生型小鼠的一般情况和在特异性黏膜免疫后 好的。在ab缺乏的小鼠身上进行的类似分析将提供证据证明 Gd T细胞可以直接支持IgA的产生，或者更确切地说，这表明它们 主要通过激活ab T细胞发挥作用。收养转移 实验将证实abT细胞的必要性，并支持这样的想法 Gd T细胞的调节作用可能是通过调节ab T细胞来实现的。在AIM 2、gd T细胞对CD_4阳性ab T细胞的调节作用 产生IgA的分析将直接在双室或共培养中进行 体外实验，并辅以T细胞转移实验 体内TCR基因敲除小鼠。Aim 3通过以下方式阐述了表达式和函数 可能直接介导细胞表面分子B71/2的SMG gd T细胞 与ab T细胞的相互作用决定了它们的细胞因子谱。目标4 研究了一种鼻腔免疫模型，以产生粘膜IgA反应 以及全身(即血清和脾)低反应性，即， 鼻腔耐受性。然后将Gd缺陷小鼠与野生型小鼠进行比较 进一步证明gd T细胞在维持粘膜功能中的作用 全身性低反应环境下的反应和鼻腔GTD 来自耐受小鼠的细胞将被用来确定它们是否可以逆转 相似耐受性的脾(系统)ab T细胞的低反应性 动物。在Aim 5中，IL-7在肿瘤的发生和增殖中的作用 在IL-7缺陷或IL-7R缺陷小鼠中将检测到SMG gd T细胞， 由于IL-7最近被证明是一组gd子集的生长因子 表达其受体的T细胞。

DESCRIPTION (Adapted from the Applicant's abstract): This is the competing renewal of an RO1 grant examining the mucosal immune response that is responsible for the induction and regulation of IgA B-cell responses. The work focuses on the murine oral/nasal cavity (e.g., submandibular gland, SMG). Previous work has shown that Th2 cell derived IL-5 and IL-6 are of particular importance for inducing sIgA positive B cells to differentiate into IgA producing plasma cells, and high levels of mRNA for these cytokines as well as IFN-gamma were reported by the investigator from SMG alpha beta (ab) TCR CD4 positive T cells. The current application proposes to examine the role of gamma delta (gd) T cells in mucosal immune responses. This direction is based on the fact that gd T cells are known to be localized to mucosal sites where they constitute approximately 50 percent of IEL in the intestine, and the investigator has found that approximately 20 percent of the T cells in the SMG are gd T cells. The investigator has shown by ELISPOT assay that the dominant Ig producing cells in SMG were IgA indicating that SMG are an IgA effector tissue, like the intestinal LP. The SMG gd T cells included IL-4 and IL-5 producing cells but not IL-4 producers. Moreover, the investigator has found in very recently completed work that gd deficient (knock out) mice have impaired mucosal IgA responses. Besides the clear role for the mucosal immune system in the production of sIgA, mucosally induced tolerance is an important phenomenon in which orally administered antigens may produce mucosal IgA responses but systemic hyporesponsiveness. The investigator has shown that mucosal gd T cells from the intestinal epithelium of orally tolerized mice can abrogate antigen-specific unresponsiveness upon adoptive transfer of gd T cells. Studies from others have shown that gd T cells can down regulate IgE responses after orally administered antigens. The investigator proposes that gd T cells may play an important regulatory role in mucosal IgA and E responses at mucosal sites through cytokine production and through interactions with ab T cells. In the first aim, SMG mononuclear cells will be examined by ELISPOT assay for IgA producing cells in gd deficient and wild type mice in general and following specific mucosal immunization with TT. Similar analyses in ab deficient mice will provide evidence for whether gd T cells can support IgA production directly, or rather suggest that they act primarily through the activation of ab T cells. Adoptive transfer experiments will confirm the need for ab T cells and support the idea that the regulatory role of gd T cells may be mediated upon ab T cells. In Aim 2, the effects of gd T cells as regulators of CD4 positive ab T cells in producing IgA will be analyzed directly in double chamber or in co-culture experiments in vitro and complemented by T cell transfer experiments using TCR knockout mice in vivo. Aim 3 addresses the expression and function by SMG gd T cells of cell surface molecules B71/2 that may mediate direct interactions with ab T cells that determine their cytokine profiles. Aim 4 examines a model of intranasal immunization to produce mucosal IgA responses along with systemic (i.e., serum and spleen) hyporesponsiveness, that is, nasal tolerance. Then gd deficient versus wild type mice will be compared to further demonstrate the role of gd T cells in maintaining mucosal responses in the setting of systemic hyporesponsiveness, and nasal gd T cells from tolerized mice will be used to determine if they can reverse the hyporesponsiveness of spleen (systemic) ab T cells from similarly tolerized animals. In Aim 5, the role of IL-7 in the development and proliferation of SMG gd T cells will be determined in IL-7 deficient or IL-7R deficient mice, since IL-7 has recently been shown to be a growth factor for a subset of gd T cells that express its receptor.