IDENTIFICATION OF HIV PROTEINS THAT STIMULATE MONOKINE SECRETION

刺激单核因子分泌的 HIV 蛋白的鉴定

• 批准号: 3811210
• 负责人: K CLOUSE-STREBEL
• 金额: --
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3811210
• 关键词: HIV infections; chimeric proteins; genetic manipulation; human immunodeficiency virus; human tissue; monokines; protein engineering; secretion; virus antigen; virus envelope; virus protein; virus replication

We have previously demonstrated that select viral antigens, including human immunodeficiency virus (HIV) itself, can stimulate human macrophages (MO) to secrete monokines capable of upregulating HIV expression in chronically infected human cells. Subsequent investigations to determine which HIV proteins were capable of stimulating monokine production revealed that the natural gpl2O envelope protein could mediate this response, whereas the natural capsid protein could not. Our recent analyses utilizing recombinant HIV proteins demonstrate that proteins expressed in baculovirus (including gpl6O and gpl2O envelope proteins and p66 reverse transcriptase; MicroGeneSys, Inc.) are unable to stimulate HIV-enhancing monokine activity. Similarly, a recombinant gpl2O fusion protein (Genentech, Inc.) expressed in Chinese hamster ovary cells is unable to induce monokine production. However, a full-length recombinant gpl2O expressed in CHO cells (Cell Tech, Inc.) is capable of stimulating monokine production. Monokine secretion caused by natural gpl2O or this latter recombinant can not be attributed to contaminating endotoxin. Furthermore, the monokine secretion caused by natural gpl2O can be blocked using either soluble CD4 or the recombinant gpl2O fusion protein. Thus, the HIV envelope glycoprotein may indirectly augment HIV expression by stimulating monokine secretion. However, subtle differences may exist in gpl2O preparations which would influence this effect on human MO.

我们之前已经证明，选择的病毒抗原，包括人类抗原， 免疫缺陷病毒（HIV）本身可以刺激人类巨噬细胞（MO） 分泌能够上调HIV表达的单核因子， 被感染的人体细胞随后的调查，以确定哪些艾滋病毒 蛋白质能够刺激单核因子的产生表明， 天然gp 120包膜蛋白可以介导这种反应，而 天然衣壳蛋白则不能。我们最近的分析利用重组 HIV蛋白表明杆状病毒（包括 gp 160和gp 120包膜蛋白和p66逆转录酶; MicroGeneSys，Inc.）无法刺激HIV增强单核因子 活动类似地，重组gpl 20融合蛋白（Genentech，Inc.） 在中国仓鼠卵巢细胞中表达不能诱导单核因子 生产然而，在CHO细胞中表达的全长重组gp 120 (Cell Tech，Inc.）能够刺激单核因子的产生。单核因子 由天然gp 120或后者重组体引起的分泌不能被 归因于污染的内毒素。此外，单核因子分泌 由天然gp 120引起的免疫排斥反应可以用可溶性CD 4或 重组gp 120融合蛋白。因此，HIV包膜糖蛋白可以 通过刺激单核因子分泌间接增加HIV表达。 然而，gp 120制剂中可能存在细微的差异， 对人类MO的影响。