INTERACTIONS OF HUMAN RETROVIRUSES WITH HEMATOPOIETIC AND ADHERENT CELLS

这个项目的目标是更好地了解主持人的影响 细胞和其他辅助因素对人类逆转录病毒复制和 发病机制。具体地说，我们已经表明HTLV-I/II具有更广泛的 宿主范围比CD4+T细胞与巨噬细胞、脑小胶质细胞和CD3- 自然杀伤细胞在体外也可以通过不含细胞的Vitrus感染。一 有证据的大颗粒淋巴细胞(LGL)白血病患者 对人T细胞白血病/淋巴瘤病毒(HTLV)感染进行了研究。克隆 扩增产物测序结果显示HTLV-II Poll和Px 病人DNA中的序列。然而，我们的数据显示，大多数患有 LGL白血病不感染典型的HTLV-I或HTLV-II。这个 患者血清对HTLV-I/II Gag蛋白p24和T0的频繁反应性 然而，环境蛋白p21e表明，缺失或变异形式的 HTLV-I/II可能与LGL白血病相关。HTLV-I的能力 产生传染性病毒粒子的分子克隆以前从未被 演示了。因此，起源于成人T细胞的HTLV-I前病毒 一名白血病患者，被克隆到一个质粒载体上，并指定了PC- HTLV。原发和继发T细胞向初级T细胞的无细胞传播 证明这是HTLV-I的一个感染性分子克隆。使用 关于HIV-I感染，现在很明显，I型患者中，有一种 生殖性感染和潜伏性感染之间的平衡。我们是 研究这种均衡的机制(S)使用 单核/巨噬细胞，艾滋病患者HIV-I感染的储存库，AS 模型单元格类型。在潜伏感染的单核细胞系THP-1中， 5-氮杂胞苷暴露后形成感染性病毒。既然是这样 表明细胞或病毒甲基化在HIV中起作用 复制，我们发现了3-去氮核苷对HIV的抗病毒作用 S-腺苷同型半胱氨酸(ADOHcy)水解酶的有效抑制剂， 细胞甲基化过程中的关键酶。出乎意料的是，在 配对AZT治疗前后治疗指标的比较 治疗HIV-I分离株，DZNep显示出3-到40倍的增长 它对耐AZT的HIV-I分离物的效力。此外，来自 无症状的血清阳性者含有潜伏的艾滋病毒。之后 这些单核细胞与Con A活化的HIV T细胞共培养 阴性的正常供者，这些潜伏的单核细胞培养表达病毒。在……里面 研究T细胞衍生的细胞因子，我们发现IL-4和IL-13，尽管 与调节单核细胞功能密切相关，可有分化 对单核细胞中HIV表达的影响。

The goal of this project is to better understand the effects of the host cell and other co-factors on human retroviral replication and pathogenesis. Specifically, we have shown that HTLV-I/II has a broader host range than CD4+ T-cells with macrophages, brain microglia and CD3- natural killer cells also infectable in vitro with cell-free vitrus. One patient with large granular lymphocyte (LGL) leukemia with evidence of human T-cell leukemia/lymphoma virus (HTLV) infection was studied. Cloning and sequencing of amplified products showed the HTLV-II pol and pX sequences in patient DNA. However, our data show that most patients with LGL leukemia are not infected with prototypical HTLV-I or HTLV-II. The frequent reactivity of patient sera to HTLV-I/II gag protein p24 and to env protein p21e, however, suggests that a deleted or variant form of HTLV-I/II may be associated with LGL leukemia. The ability of HTLV-I molecular clones to produce infectious virions has not previously been demonstrated. Thus, an HTLV-I provirus originating from an adult T-cell leukemia patient, was cloned into a plasmid vector and designated pCS- HTLV. Primary and secondary cell-free transmissions into primary T-cell demonstrated that this is an infectious molecular clone of HTLV-I. With regard to HIV-I infection, it is now clear that i patients, there is an equilibrium between productive infection and latent infection. We are studying the mechanism(s) for this equilibrium using the monocyte/macrophage, a reservoir of HIV-I infection in AIDS patients, as the model cell type. In latently infected THP-1, a monocytoid cell line, infectious virus was made after 5-azacytidine exposure. Since this suggests that either cellular or viral methylation plays a role in HIV replication, we found an antiviral effect on HIV by 3-Deaza-nucleoside compounds, potent inhibitors of S-adenosylhomocysteine (AdoHcy) hydrolase, a key enzyme in cellular methylation processes. Unexpectedly, in comparing the therapeutic indice between the paired pre- and post-AZT treatment HIV-I isolates, DZNep displayed an increase of 3- to 40-fold in its potency against AZT-resistant HIV-I isolates. Also, monocytes from asymptomatic seropositive individuals contained latent HIV. After coculture of these monocytes with Con A-activated T-cells from HIV negative normal donors, these latent monocyte cultures expressed virus. In studying T-cell derived cytokines, we found that IL-4 and IL-13, although closely related in modulating monocyte function, can have divergent effects on HIV expression in monocytes.