PATHOGENIC EFFECTS OF HUMAN RETROVIRUSES ON HEMATOPOIETIC AND ADHERENT CELLS

这个项目的目标是更好地了解东道主的影响 细胞和其他辅助因素对人类逆转录病毒复制和 发病机制。日本血吸虫感染性分子克隆的研制 人T细胞白血病淋巴瘤病毒I型(HTLV-1) 无细胞感染导致细胞因子依赖性转化和 不需要HTLV-I的P12和P30开放阅读框 转型。现在很明显，HTLV，HIV和人类泡沫病毒， 生殖性感染和潜伏性感染之间存在着平衡。 在艾滋病患者中，我们和其他人发现这种平衡可以 因免疫刺激和甲基化而不安。我们正在研究 使用T细胞改变这种平衡的致病后果 非细胞/巨噬细胞。无症状HIV+个体的单核细胞 含有潜伏的艾滋病毒。将这些单核细胞与刀豆蛋白A共培养后- 来自HIV阴性的正常捐赠者的激活的T细胞，这些单核细胞 表达的病毒。在潜伏感染的THP-1中，产生了传染性病毒 在5-氮胞苷暴露后。因为这表明细胞 甲基化在HIV复制中起作用，我们研究了HIV 感染可能导致细胞基因的异常甲基化。一个 干扰素-γ的从头甲基化显著增加 在这些基因中发现了与表达降低相关的启动子 受到严重感染的细胞。一种CD4+类淋巴细胞系--JMO，其表达 利用表达载体稳定地表达了干扰素-γ。 反义定向构建含DNA MTase基因的载体(TMH)。 核糖核酸酶保护分析表明， MTase在表达反义MTase的JMO中的表达南方 分析显示JMO中的干扰素-γ启动子半甲基化 表达亲本neo构建体的细胞株，而JMO-TMH 完全低甲基化。细胞系产生干扰素-γ的研究 构建表达亲本载体。在急性感染后 爱滋病毒。 艾滋病标题：艾滋病毒复制的负面调控-致病效应

The goal of this project is to better understand the effects of host cells and other co-factors on human retroviral replication and pathogenesis. After development of the infectious molecular clone of Human T cell Leukemia Lymphoma virus type I (HTLV-1), we determined that cell-free infrection results in cytokine dependent transformation and that p12 and p30 open reading frames of HTLV-I are not needed for transformation. It is now clear that HTLV, HIV and Human Foamy Virus, there is a balance between productive infection and latent infection. In AIDS patients, we and others have found that this equilibrium can upset by immune stimulation and methylation. We are studying the pathogenic consequences of altering this balance using T cells and nocyte/macrophages. Monocytes from asymptomatic HIV+ individuals contained latent HIV. After coculture of these monocytes with Con A- activited T-cells from HIV negative normal donors, these monocytes expressed virus. In latently infected THP-1, infectious virus was made after 5-azacytidine exposure. Since this suggests that cellular methylation plays a role in HIV replication, we studied whether HIV infection could result in aberrant methylation of cellular genes. A significant increase in the de novo methylation of the IFN-gamma promoter, which correlated with decreased expression, was seen in these acutely infected cells. a CD4+ lymphoid cell line-JMO, which expresses IFN-gamma constitutively, was stablely transfected using expression vectors containingf DNA MTase cDNA in the antisense orientation (TMH). Rnase protection analyses demonstrated a significantly lover level of MTase expression in the JMO expressing antisense MTase. Southern anaylsis showed hemimethylation of the IFN-gamma promoter in the JMO cell line expressing the parental neo construct while JMO-TMH was completely hypomethylated. IFN-gamma production in cell line constitutively expressing the parental vector. After acute infection of HIV. AIDS Title: Negative Regulation of HIV replication - Pathogenic Effects