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复制的潜在靶标，合成肽T134和T140特别抑制X4 HIV-1感染，因为它们充当CXCR4拮抗剂以阐明其机制。为了了解 CXCR4 拮抗剂的作用，我们在细胞培养物中产生了 T134 耐药性 HIV (trHIV-1NL4-3)，并逐渐增加浓度通过MTT测定法和MAGI测定法评估了几种CXCR4拮抗剂、SDF-1和v MIP II（其在KSHV/HHV8中编码为趋化因子样肽）的抗HIV活性。高浓度CXCR4拮抗剂对R5的作用。还研究了 HIV -1 trHIV-1NL4-3 糖蛋白区域的氨基酸突变以及其他抗 HIV 化合物的交叉耐药性。研究结果显示，高浓度的 CXCR4 拮抗剂增加了 CCR5 表达和 R5 HIV-1 感染性，CXCR4 拮抗剂和 SDF-1 也增加了 NF-kB 活性和病毒。经处理的细胞中 trHIV-1NL4-3 的转录减少了 15 倍，并且还降低了对其他 CXCR4 拮抗剂的敏感性， T140、AMD3100 和 ALX40-4C 和 SDF-1 然而，可抑制 X4 和 R5 HIV-1 感染的 vMIP II 仍然对 trHIMV-1NL4-3 配体敏感。 CCR5 低分子拮抗剂 TAK-779 也无法影响 trHIV-1NL4-3 的感染。 trHIV-1NL4-3 不仅在 gp120 的 V3 环中含有多个突变，而且在 gp120 的 V1、V2 和 V4 结构域中也含有多个突变，因此，对 T134 的抗性可能是通过 X4 HIV-1.2) 的包膜糖蛋白中的氨基酸取代而赋予的。口腔上皮细胞和TPA刺激HHV8整合BCBL-1细胞，并通过PCR方法在显微镜下观察HHV8基因组DNA和mRNA。在培养细胞中观察到细胞病变效应，但没有检测到上皮细胞中 HHV8 感染的证据。