FUNCTION AND REGULATION OF VLA MOLECULES ON T CELLS

T细胞在发育过程中和成熟后， VLA蛋白家族中的受体与其他细胞相互作用， 细胞外基质从初步结果来看，VLA 蛋白质不仅是底物或靶结构的受体 参与细胞定位和迁移，而且还可以直接 有助于通过T细胞发出信号，并且可以进行功能调节 对来自T细胞的信号作出反应。 这项研究的长期目标是 是理解VLA蛋白质之间惊人的双向联系， T细胞激活事件，这样我们就可以有一个更好的整体 了解T细胞的发育和功能。 为了实现这一目标，[1] 我将研究T细胞活化（通过抗原、抗体、酯）的结果 在VLA蛋白粘附于细胞和细胞外的快速改变中， 基质配体 特别是，我们将探讨一些VLA函数是如何 可能同时在相反的方向上调节。[2)将 分析VLA蛋白配体或抗VLA单克隆抗体（MAb） 它们本身可以直接或间接地促进T细胞活化， 通过增殖、钙流和淋巴因子分泌来测量。 的 目的#1和#2中提到的研究将利用正常T细胞克隆， 细胞系、T细胞受体（TCR）阴性突变体和TCR重建 突变体，以评估VLA蛋白和TCR之间的连接。[3]在 此外，使用可用的α和β亚基cDNA，我们将删除和 交换VLA蛋白α和β亚基胞质结构域，然后 将测试变体VLA（在T细胞上表达）的i）它们的能力 ii）它们促进T细胞增殖的能力， activation. 因此，我们将确定胞质结构域在细胞内的作用。 T细胞和VLA蛋白之间的明显双向信号传导。[4]美国 为了研究胸腺细胞上的VLA蛋白，我们将观察它们的阶段- 特异性表达、配体结合功能以及这些功能的调节 功能协调发展的[5]此外，为了充分研究小鼠T细胞上的VLA蛋白， 胸腺细胞，我们将产生急需的抗小鼠VLA蛋白单抗。 这些实验将共同提供重要的见解， VLA蛋白的各种细胞和细胞外基质配体可以帮助 来指导T细胞发育和功能的有序程序。

T cells, both during development and after maturation, utilize adhesion receptors in the VLA protein family to interact with other cells and with extracellular matrix. From preliminary results it now appears that VLA proteins are not only receptors for substrates or target structures involved in cell positioning and migration, but also can directly contribute to signalling through T cells, and can be functionally modulated in response to signals from T cells. The long term goal of this research is to comprehend this striking two-way connection between VLA proteins and T cell activation events, so that we can have a better overall understanding of T cell development and function. To this goal, [1] we will look at how T cell activation (by antigen, antibodies, esters) results in rapid alterations in VLA protein adhesion to both cell and extracellular matrix ligands. In particular, we will explore how some VLA functions are possibly regulated in opposite directions at the same time. [2) will analyze how VLA protein ligands or anti-VLA monoclonal antibodies (MAb) themselves can directly or indirectly facilitate T cell activation, as measured by proliferation, calcium flux, and lymphokine secretion. The studies mentioned in Aims #1 and #2 will utilize normal T cell clones and lines, T cell receptor (TCR) negative mutants, and TCR reconstituted mutants to assess the connection between VLA proteins and the TCR. [3] In addition, using available alpha and beta subunit cDNA's, we will delete and exchange VLA protein alpha and beta subunit cytoplasmic domains, and then variant VLA's (expressed on T cells) will be tested for i) their ability to be functionally regulated, and ii) their ability to facilitate T cell activation. Thus we will determine the role cytoplasmic domains in the apparent bi-directional signalling between T cells and VLA proteins. [4] To investigate VLA proteins on thymocytes, we will look at their stage- specific expression, ligand-binding functions, and the regulation of these functions.[5] Also, to adequately study VLA proteins on mouse T cells and thymocytes we will generate urgently needed anti-mouse VLA protein MAb. Together these experiments will give important insights into how the various cellular and extracellular matrix ligands of VLA proteins can help to direct an orderly program of T cell development and function.