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

人星形胶质细胞在体内和体外均可被HIV-1感染。 然而，HIV-1结构和非结构蛋白的量 与巨噬细胞或小胶质细胞相比， 大脑中的细胞。 在使用人胎儿星形胶质细胞的体外模型中， 感染或转染后数周，与 未感染的淋巴细胞或用细胞因子TNF-α刺激， IL 1-β将增加这种细胞类型的病毒产量。在 目前的工作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。 我们还显示了在体内感染 星形胶质细胞与HIV-1表达的非结构蛋白，nef。 这 其他研究人员独立证实了观察结果， 儿科病例星形胶质细胞感染在发病机制中的作用 HIV-1相关脑病的研究仍在继续。 此外，HIV-1 似乎建立了持续感染的基质细胞的 淋巴组织这种感染与 星形胶质细胞正在分子水平上靶向NF-κ B进行研究 诱导和结合HIV-1 LTR。 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。

Human astrocytes can be infected with HIV-1 both in vivo and in vitro. However, the amount of HIV-1 structural and nonstructural protein production is low compared to that of the macrophage or the microglial cell in the brain. In an in vitro model using 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. In the current work 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. We have also shown in vivo infection of astrocytes with HIV-1 expressing the nonstructural protein, nef. This observation was confirmed independently by other investigators in pediatric cases. The role of the infected astrocyte in the pathogenesis of HIV-1-associated encephalopathy continues to be studied. Also, HIV-1 appears to establish a persistent infection in the stromal cells of the lymphoid tissues. The similarities of this infection to that of the astrocyte is being studied at the molecular level targeting NF-kB induction and binding to the HIV-1 LTR. 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.