MECHANISMS OF HEMATOLOGIC ABNORMALITIES IN AIDS

说明(改编自《调查员摘要》)： 细胞因子、生长因子、细胞外基质蛋白及最新证据 病毒蛋白对多能干细胞分化的影响 申请人假设病毒Tat和/或Nef蛋白可能会影响 造血祖细胞的全面增殖和分化 通过改变基质中各种细胞因子的产生来影响细胞 细胞和/或诱导对CD34+细胞的直接抑制作用。至 检验这一假设，私家侦探建议调查TAT的影响 和/或Nef蛋白，直接或通过共培养技术 利用HeLa-Tat和/或HeLa-nef稳定表达的细胞系 原代CD34+细胞、基质细胞和转化的祖细胞， K562和HL-60。具体来说，将调查以下目标：(1) CD34+细胞、K-562、HL-60细胞的增殖分化 在存在和不存在未经处理和预处理的基质细胞的情况下 (2)病毒蛋白对信号的影响 信号转导途径；(3)基质细胞释放的细胞因子水平 细胞和相应mRNA水平的监测；(4)表达 原癌基因和细胞表面受体的调节；(5) 多种转录因子通过直接蛋白-蛋白发挥作用 相互作用；以及(6)可能导致病毒的机制 蛋白质诱导的祖细胞凋亡途径的调节。 申请人相信，拟议的实验将提供一个 深入了解可能涉及艾滋病毒诱导的机制 造血抑制。

DESCRIPTION (Adapted from the Investigator's Abstract): Effects of cytokines, growth factors, extracellular matrix proteins and recent evidence of viral proteins on pluripotent stem cell differentiation has led the applicant to hypothesize that viral Tat and/or Nef proteins may affect the overall proliferation and differentiation of the hematopoietic progenitor cells via alteration of production of the various cytokines in the stromal cells and/or induction of a direct inhibitory effect on the CD34+ cells. To test this hypothesis, the P.I. proposes to investigate the effects of Tat and/or Nef proteins, either directly or by the co-culture techniques utilizing the HeLa-tat and/or HeLa-nef stably transfected cell lines on the primary CD34+ cells, the stromal cells and the transformed progenitor cells, K562 and HL-60. Specifically, following aims will be investigated: (1) proliferation and differentiation of CD34+ cells, and K-562 and HL-60 cells in the presence and absence of stromal cells both untreated and pretreated with the viral proteins; (2) the effect of the viral proteins on signal transduction pathways; (3) the levels of cytokines released by the stromal cells and monitoring of the corresponding mRNA levels; (4) the expression of protooncogenes and of the cell surface receptors; (5) the modulation of the function of various transcription factors via direct protein-protein interactions; and (6) the mechanisms that may be responsible for the viral protein-induced modulation of apoptotic pathways in the progenitor cells. The applicant believes that the proposed experiments will provide an in-depth understanding of the mechanisms that may be involved in HIV-induced hematopoietic suppression.