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

HHV8感染细胞和口腔上皮细胞表达生长因子的研究

基本信息

批准号：
10470392
负责人：
NAKASHIMA Hideki
金额：
$ 7.17万
依托单位：
St. Marianna University School of Medicine (2001)Kagoshima University (1998-2000)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-10470392/
关键词：
T134 T140 AMD3100 CXCR4 antagonists KSHV HHV8 vMIP-II BCBL-1 ベツリン酸誘導体 YK-FH312 MAGIアッセイ CCR5 CXCR4 交差耐性

项目摘要

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

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特异性地抑制X4型HIV-1感染，因为它们作为CXCR4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T134抗性HIV （trHIV-1NL4-3）。用MTT法和MAGI法评价了在KSHV/HHV8中编码为趋化因子样肽的CXCR4拮抗剂SDF-1和v MIP II的抗hiv活性。我们还研究了高浓度CXCR4拮抗剂对R5型HIV-1的作用。对trHIV-1NL4-3糖蛋白区氨基酸突变进行了测序，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R5的HIV-1传染性。CXCR4拮抗剂和SDF-1也在处理细胞中增加NF-kB活性和病毒转录。trHIV-1NL4-3对其他CXCR4拮抗剂（T140、AMD3100、ALX40-4C和SDF-1）的敏感性降低了15倍。然而，vvmip II对trHIMV-1NL4-3仍然敏感，对X4和R5型HIV-1感染均有抑制作用。CCR5配体、RANTES和MIP-1α以及CCR5低分子拮抗剂TAK-779都不能影响trHIV-1NL4-3的感染。trHIV-1NL4-3不仅在gp120的V3环上，而且在gp120的V1、V2和V4结构域上都有多个突变。因此，对T134的抗性可能与X4 hiv -1包膜糖蛋白的氨基酸取代有关。1.2)口腔上皮细胞与TPA刺激HHV8整合BCBL-1细胞共培养，并在显微镜下观察。采用PCR法检测HHV8基因组DNA和mRNA。在培养细胞中观察到细胞致病作用，但在上皮细胞中未检测到HHV8感染的证据。少