Study for Growth Factors which Expressed in HHV8 Infected Cells and Oral Epithelial Cells

1)趋化因子受体CXCR4和CCR5被认为是抑制HIV-1复制的潜在靶点。合成肽T134和T140特别能抑制X4HIV-1感染，因为它们是CXCR4的拮抗剂。为了阐明CXCR4拮抗剂的作用机制，我们在逐渐增加T134浓度的细胞培养中产生了耐T134的HIV(trHIV-1NL4-3)。用四甲基偶氮唑盐比色法和MAGI比色法检测了KSHV/HHV8编码的趋化素样多肽SDF-1和v MIP II的抗HIV活性。研究了高浓度CXCR4拮抗剂对R5型HIV-1的作用。测定了trHIV-1NL4-3糖蛋白的氨基酸序列，并研究了其他抗HIV化合物对trHIV-1NL4-3的交叉耐药性。结果表明，高浓度的CXCR4拮抗剂和SDF-1一样，增加了CCR5的表达和R5HIV-1的感染力。CXCR4拮抗剂和SDF-1也包括…处理组细胞中的核因子-kB活性和病毒转录水平明显降低。TtrHIV-1NL4-3减少了15倍，并降低了对其他CXCR4拮抗剂T140、AMD3100和ALX40-4C以及SDF-1的敏感性。但同时抑制X4和R5型HIV-1感染的vMIP II对trHIMV-1NL4-3仍然敏感。CCR5、RANTES和MIP1α的配体以及CCR5小分子拮抗剂TAK779都不能影响trHIV-1NL4-3的感染。TrHIV-1NL4-3不仅在gp120的V3环上存在多个突变，而且在V1、V2和V4区域也存在突变。2)口腔上皮细胞与TPA刺激整合的HHV8整合的BCBL-1细胞共培养，并在显微镜下观察。用聚合酶链式反应方法对HHV8基因组DNA和mRNA进行检测。在培养的细胞中观察到了细胞病变效应，但在上皮细胞中没有检测到HHV8感染的证据。较少

1) The chemokine receptors, CXCR4 and CCR5, are considered to be potential targets for the inhibition of HIV-1 replication. Synthetic peptides, T134 and T140, inhibited X4 HIV-1 infection specifically because they acted as CXCR4 antagonists. To clarify the mechanisms of action of CXCR4 antagonists, we generated T134-resistant HIV (trHIV-1NL4-3) in a cell culture with gradually increasing concentrations of T134. Anti-HIV activities of several CXCR4 antagonists, SDF-1 and v MIP II which coded in KSHV/HHV8 as a chemokine like peptide, were evaluated by MTT assay and MAGI assay. The effects of high concentrations of CXCR4 antagonists against R5 HIV-1 were also investigated. Amino acids mutations of trHIV-1NL4-3 glycoprotein region were sequenced and cross resistancies of other anti-HIV compounds against trHIV-1NL4-3 were studied. As the results, high concentrations of CXCR4 antagonists increased the CCR5 expression and R5 HIV-1 infectivity as did SDF-1. CXCR4 antagonists and SDF-1 also inc … More reased NF-kB activity and viral transcription in the treated cells. The t trHIV-1NL4-3 reduced 15-fold less and also reduced sensitivities against other CXCR4 antagonists, T140, AMD3100 and ALX40-4C, and SDF-1. However, vMIP II which could inhibit both X4 and R5 HIV-1 infection, was still sensitive against trHIMV-1NL4-3. Neither ligands of CCR5, RANTES and MIP-1α, nor a CCR5 low molecular antagonist, TAK-779, were able to influence the infection of trHIV-1NL4-3. The trHIV-1NL4-3 contained several mutations in the not only V3 loop but also V1, V2 and V4 domains of gp120. Thus, resistance to T134 may be conferred by amino acid substitutions in the envelope glycoprotein of X4 HIV-1.2) The coculture of oral epithelial cells and TPA stimulate HHV8 integrated BCBL-1 cells were carried out and the observed under microscopically. HHV8 genomic DNA and mRNA were also investigated by a PCR method. The cytopathogenic effects were observed in cultured cells but no evidence of HHV8 infection in epithelial cells were detected. Less