IDENTIFICATION OF HIV PROTEINS THAT STIMULATE MONOKINE SECRETION

我们以前已经证明，选择的病毒抗原，包括 人类免疫缺陷病毒（HIV）本身可以刺激人体巨噬细胞 (MO)分泌能够上调HIV表达的单核因子， 慢性感染的人体细胞 随后的调查确定 哪些HIV蛋白能够刺激单核因子的产生 揭示了天然的gp 120包膜蛋白可以介导这一过程。 反应，而天然衣壳蛋白不能。 我们发现 天然gp 120刺激MO释放TNF-α、IL-1 β、IL-6和GM-1， 这种作用可被可溶性CD 4阻断。 分析 利用重组HIV蛋白证明， 在杆状病毒中表达，包括gp 160和gp 120，不能 刺激单核因子分泌。 类似地，重组gp 120融合蛋白 在中国仓鼠卵巢（CHO）细胞中表达的蛋白不能 以这种能力发挥作用。 然而，全长重组gp 120 在CHO细胞中表达的重组蛋白能够刺激单核因子的产生。 的 天然和全长CHO细胞衍生刺激能力 gp 120通过在100摄氏度下加热而被消除，并且可以被阻断。 过量CHO细胞衍生的重组gp 120融合蛋白。 以来 杆状病毒表达的gp 120和CHO细胞衍生的重组融合体 这些测试结果表明，HIV gp 120与CD 4的结合 MO表面上的CD 4本身可能不足以刺激 单核因子分泌

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 gp120 envelope protein could mediate this response, whereas the natural capsid protein could not. We found that natural gp120 stimulates MO to release TNF-alpha, IL-1beta, IL-6 and GM- CSF, and this effect could be blocked with soluble CD4. Analysis utilizing recombinant HIV proteins demonstrate that envelope proteins expressed in baculovirus, including gp160 and gp120, are unable to stimulate monokine secretion. Similarly, a recombinant gp120 fusion protein expressed in Chinese hamster ovary (CHO) cells is unable to function in this capacity. However, a full-length recombinant gp120 expressed in CHO cells is capable of stimulating monokine production. The stimulatory capacity of both natural and full-length CHO-cell derived gp120 was eliminated by heating at 100 degrees C, and could be blocked with excess CHO-cell derived recombinant gp120 fusion protein. Since the baculovirus-expressed gp120 and the CHO-cell derived recombinant fusion protein bind to CD4, these test results suggest that HIV gp120 binding to CD4 on the MO surface may of itself be insufficient for stimulation of monokine secretion.