HIV-1 INFECTION IN FETAL BRAIN CELL CULTURES AND PEDIATRIC AIDS BRAIN TISSUE

我们建立了一种人HIV-1潜伏感染的体外模型 胎儿星形胶质细胞 感染或转染后数周， 与未感染的淋巴细胞共培养或用 细胞因子TNF-α和IL-1-β将增加病毒的产生， 细胞类型。我们已经证明佛波醇12-肉豆蔻酸酯13-乙酸酯 (PMA)也增加了HIV-1 p24的产生， 星形胶质细胞采用电泳迁移率变动分析（EMSA）， 结合使用特异性抗体的超移位研究，我们 表明PMA，像TNF-α一样，增加NF-κ B的p50/p65形式。 KB. 此外，我们还表明，蛋白激酶抑制剂H7 抑制PMA和TNF-α相关的HIV-1表达增加， 当它对相关的细胞几乎没有抑制作用时， p50/p65 NF-kB增加。 因此，除非p50/p65 NF-kB或其结合 受H7影响的方式无法通过EMSA解决， 这种形式NF-kB的增加并不总是足以增加HIV-1 从星形胶质细胞表达。 该实验室还在研究特定RNA酶的能力， 抑制HIV-1在淋巴细胞系中的增殖。 Onconase 和牛精液核糖核酸酶被证明可以阻断HIV-1的感染， 生产力感染细胞系。 此块显示在 RNase活性的细胞内机制。 除了在受感染的星形胶质细胞培养物中调节NF-κ B外， 病毒Rev蛋白似乎也是细胞控制的靶。 使用EMSA分析，星形胶质细胞产生结合RRE-rev的因子， 复杂. 该因子还对细胞因子和PMA刺激做出反应。 NF-kB和rev结合因子似乎都受到限制， 与对HIV-1感染高度敏感的细胞相比， 这些数据指出了HIV-1在脑内潜伏感染的机制 涉及降低生产性必需的细胞因子水平 HIV-1增殖。 星形胶质细胞作为HIV-1储存库的作用 正在使用体外感染模型研究大脑中的病毒。

We have established an in vitro model of latent HIV-1 infection in human fetal astrocytes. Several weeks following infection or transfection, cocultivation with uninfected lymphocytes or stimulation with the cytokines TNF-alpha and IL1-beta will increase viral production from this cell type. We have demonstrated that phorbol 12-myristate 13-acetate (PMA) also increases HIV-1 p24 production from the primary human astrocyte. Using electrophoretic mobility shift assay (EMSA) in combination with supershift studies using specific antibodies, we demonstrated that PMA, like TNF-alpha increases the p50/p65 form of NF- kB. Furthermore, we also showed that the protein kinase inhibitor H7 inhibits PMA and TNF-alpha associated increases in HIV-1 expression at a time when it has little to no inhibitory effect on the associated increases in p50/p65 NF-kB. Thus, unless p50/p65 NF-kB or its binding is affected by H7 in a manner that cannot be resolved by EMSA, an increase in this form of NF-kB is not always sufficient to increase HIV-1 expression from the astrocyte. The laboratory is also investigating the ability of specific RNases to inhibit the multiplication of HIV-1 in lymphocyte cell lines. Onconase and bovine seminal RNase were shown to block infection of HIV-1 in productivity infected cell lines. This block appears from an intracellular mechanism of RNase activity. In addition to the regulation of NF-kB in infected astrocyte cultures, the viral rev protein also seems to be a target for cellular control. Using EMSA assays, astrocytes produce a factor which binds the RRE-rev complex. This factor also responds to cytokine and PMA stimulation. Both the NF-kB and the rev binding factor appear to be limited in concentration compared with cells highly susceptible to HIV-1 infection. These data point to a mechanism of HIV-1 latent infection in brain involving reduced levels of cellular factors necessary for productive HIV-1 multiplication. The role of the astrocyte as a reservoir of HIV-1 in the brain is being investigated using the in vitro model of infection.