LYMPHOCYTE/VASCULAR INTERACTIONS IN IMMUNE RESPONSES

关于控制这一现象的机制的信息很少。 抗原特异性淋巴细胞的定位。这个问题是显而易见的 了解抗原特异性淋巴细胞的稀有程度很重要 能够定位并对外来抗原作出反应或启动自身免疫 组织损伤。因此，这笔赠款的重点是审查 抗原特异性淋巴细胞定位的分子机制 抗原部位或淋巴器官引流部位 发炎。 淋巴细胞从血管系统向组织中的募集主要是 依赖于淋巴细胞与内皮细胞的相互作用。在具体目标1中， 我们建议研究CD31，一种可能的黏附分子，在 淋巴细胞从循环中向组织中迁移。CD31，a 作为免疫球蛋白超家族的一员，可能在 淋巴细胞通过血管内皮细胞的移行。我们有 开发并鉴定了目前唯一可用的单抗 抗小鼠CD31抗体为体内研究提供了独特的工具 学习。此外，我们还建立了培养体系，采用 小鼠内皮细胞和淋巴样细胞，将使我们能够解剖 该分子在淋巴细胞-内皮细胞相互作用中的作用。 通过我们开发的一种新的免疫组织化学技术，我们发现 抗原反应性淋巴细胞被选择性地保留在 受刺激淋巴器官的血管周围区域。在具体目标2中， 我们将研究调节这一过程的分子/机制。 血管周围组织中抗原反应性淋巴细胞的滞留。我们会 特征：(1)辅助细胞在影响 抗原反应性淋巴细胞的定位，以及(2)潜在的作用 β1整合素在介导淋巴细胞黏附中的作用 至细胞外基质，如血管周围组织的基底膜。 因此，这些研究将把淋巴细胞性炎症和 在组织和器官中的定位与抗原的诱导- 特定的免疫反应。这些研究的一个重要的新方面 是利用体内的实验模型。 在特定目标3中，我们将表征由以下分子合成的独特分子 抗原反应性的血管周围淋巴细胞。我们已经将一个 命名为2B6的单抗，仅识别血管周围 淋巴细胞。在抗原诱导过程中加入体外 淋巴增殖反应，高效液相纯化的2B6单抗介导了强大的 抑制效应。我们建议通过以下方法来鉴定这种分子 免疫沉淀后的SDS-PAGE，可能还有N-末端的氨基 酸分析。此外，我们将进一步研究这一点的作用 分子通过试图在体内阻断该分子的功能 使用2B6单抗。这种方法是一个更广泛关注的目标，它将 可能会对免疫反应的生理学产生新的见解，因为 解剖定位的血管周围亚群包括显著的 体内部分抗原反应性淋巴细胞。

There is a paucity of information about the mechanisms controlling the localizing of antigen-specific lymphocytes. This issue is of obvious importance to understanding how rare antigen-specific lymphocytes are able to locate and respond to foreign antigens or initiate autoimmune tissue injury. The focus of this grant, therefore is to examine the molecular mechanisms by which antigen-specific lymphocytes localize in sites of antigen or to the lymphoid organs draining sites of inflammation. Lymphocyte recruitment from vasculature into tissues is primarily dependent upon lymphocyte-endothelial interactions. In specific aim 1, we propose to examine the role of CD31, a putative adhesion molecule, in lymphocyte transmigration from the circulation into tissues. CD31, a member of the immunoglobulin superfamily, may play a key role in transmigration of lymphocytes through vascular endothelium. We have developed and characterized the only presently available monoclonal antibody against murine CD31, providing a unique tool for in vivo studies. In addition, we have established cultured systems employing murine endothelium and lymphoid cells that will allow us to dissect the role of this molecule in lymphocyte-endothelial interactions. With a novel immunohistochemical technique that we developed, we found that antigen-reactive lymphocytes are selectively retained in the perivascular region of stimulated lymphoid organs. In specific aim #2, we will investigate the molecules/mechanisms which mediate this perivascular tissue retention of antigen-reactive lymphocytes. We will characterize: (1) the role of accessory cells in affecting the localization of antigen-reactive lymphocytes, and (2) the potential role served by the Beta1 integrins in mediating the attachment of lymphocytes to extracellular matrices, such as basal lamina of the perivasculature. These studies will therefore correlated lymphocytic inflammation and localization in tissues and organs with the induction of antigens- specific immune responses. An important novel aspect of these studies is the use of in vivo experimental models. In specific aim 3, we will characterize unique molecules synthesized by antigen-reactive, perivascular lymphocytes. We have characterized a monoclonal antibody designated 2B6, which recognized only perivascular lymphocytes. When added in vitro during antigen-induced lymphoproliferative response, the HPLC-purified 2B6 mAb mediates a strong suppressive effect. We propose to identify this molecule by immunoprecipitation followed by SDS-PAGE, and possibly N-terminal amino acid analysis. Additionally, we will further examine the role of this molecule by attempting to block the function of this molecule in vivo using the 2B6 mAb. This approach is a more broadly focused aim that will likely yield new insights into the physiology of immune responses, since the anatomically localized perivascular subset comprises a significant portion of antigen-reactive lymphocytes in vivo.