THROMBOSPONDIN IN PHAGOCYTE ACTIVATION

描述：(改编自调查人员摘要)这是一个修订后的 关于继续研究TSP和ITS作用的建议 炎症反应中的受体。需要检验的假设是 中性粒细胞和单核细胞对TSP的异质性反应源于 发育过程中TSP受体(TSP-R)的差异。 特定目标1解决TSP在吞噬细胞功能中的作用 回应。人外周血单核细胞的功能研究 在继续表征PMN的同时进行-和 分化的HL-60细胞与TSP的相互作用。它也将被确定 中性粒细胞和单核细胞与TSP黏附诱导炎性细胞因子基因 表情。特定目标2使用HL-60细胞模型来测试 TSP对单核细胞和中性粒细胞行为的不同影响假说 TSP-R在分化过程中表达的差异。这个 TSP诱导或改变HL-60细胞分化的能力 被评估。类似的研究也将在单核细胞上进行。 将利用重组cDNA技术来减少表达 未分化HL-60细胞中TSP检测对HL-60细胞分化的影响 在没有配体的情况下表达TSP-R。具体目标3 继续并扩展TSP结合研究，以进一步表征 感受器。这种受体将从两者中鉴定和分离出来 中性粒细胞和单核细胞的标准生化技术及表达 克隆。利用分化的HL-60细胞，将确定是否 吞噬细胞表达蛋白多糖受体，可以与 TSP的肝素结合区(HBD)。《特定目标4》将考察 TSP-R与细胞骨架的相互作用，并确定这些 受体与G蛋白有关。

DESCRIPTION: (Adapted from investigator's abstract) This is a revised proposal for the continuation of studies on the role of TSP and its receptors in the inflammatory response. The hypothesis to be tested is that the heterogeneous responses of PMNs and monocytes to TSP result from differences in the TSP receptors (TSP-R) that occur during development. Specific Aim 1 addresses the role of TSP in phagocyte functional responses. Functional studies on human peripheral blood monocytes will be conducted while continuing the characterization of PMN- and differentiated HL-60 cell-TSP interactions. It will also be determined if PMN and monocyte adherence to TSP induces inflammatory cytokine gene expression. Specific Aim 2 uses the HL-60 cell model to test the hypothesis that TSP differentially affects monocyte and PMN behavior through differences in TSP-R expression during differentiation. The ability of TSP to induce or alter the differentiation of HL-60 cells will be evaluated. Similar studies will be undertaken on monocytes. Recombinant cDNA technology will be utilized to reduce the expression of TSP in undifferentiated HL-60 cells to evaluate HL-60 differentiation and TSP-R expression in the absence of the ligand. Specific Aim 3 continues and extends the TSP binding studies to further characterize the receptor. This receptor will be identified and isolated from both PMNs and monocytes by standard biochemical techniques and expression cloning. Using differentiated HL-60 cells, it will be determined if phagocytes express proteoglycan receptors that can interact with the heparin binding domain (HBD) of TSP. Specific Aim 4 will examine the interaction of the TSP-R with the cytoskeleton, and determine if these receptors are associated with G-proteins.