Stimulation of HIV gp120-specific cytolytic T lymphocyte responses in vitro and in vivo using a detoxified pertussis toxin vector.
Stimulation of HIV gp120-specific cytolytic T lymphocyte responses in vitro and in vivo using a detoxified pertussis toxin vector.
复制标题
使用解毒百日咳毒素载体在体外和体内刺激 HIV gp120 特异性溶细胞 T 淋巴细胞反应。
DOI：
10.1089/088922201750252016
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发表时间：
2001
期刊：
影响因子：
--
通讯作者：
Lewis,GK
中科院分区：
文献类型：
--
作者：
Carbonetti,NH;Tuskan,RG;Lewis,GK
CD 8+细胞溶解性T淋巴细胞（CTL）几乎肯定是对HIV的潜在保护性免疫应答的重要组成部分。为了测试百日咳毒素（PT）递送HIV衍生的 为了获得一种用于CTL刺激的主要组织相容性复合物（MHC）I类肽，我们构建了gp 120 P18-I10 CTL表位与PT的遗传去毒衍生物（PT 9 K/129 G）的融合物，并测定了 该融合体在体外和体内刺激gp 120特异性CTL应答的能力。与该融合蛋白孵育的抗原呈递细胞被P18-I10特异性CTLin体外裂解，该活性被证明是MHC I类限制性的。该活性被布雷菲德菌素A抑制，但不被蛋白酶体抑制剂抑制，这可能是因为PT经历了逆行 通过高尔基体细胞内转运到内质网，并将表位直接递送到新生的I类分子。用单剂量的融合物腹膜内免疫小鼠 没有佐剂的蛋白在脾中引起强烈的gp 120特异性CTL应答。该CTL应答依赖于（1）施用的融合物的剂量，（2）表位与毒素的融合物（因为融合物的抗原性），（3）表位与毒素的融合物（因为融合物的抗原性），（4）表位与毒素的融合物（因为融合物的抗原性）。 肽和毒素的共同施用没有产生应答），和（3）CD 8+细胞的活性。这些数据表明，这种解毒的PT衍生物，已经是一种许可的成分， 人的疫苗，可以代表用于刺激HIV特异性CTL应答的有用的疫苗载体分子。
CD8+cytolytic T lymphocytes (CTL) are almost certainly an important component of a potentially protective immune response to HIV. To test the ability of pertussis toxin (PT) to deliver an HIV-derived major histocompatibility complex (MHC) class I peptide for CTL stimulation, we constructed a fusion of the gp120 P18-I10 CTL epitope with a genetically detoxified derivative of PT (PT9K/129G) and assayed this fusion for its ability to stimulate a gp120-specific CTL responsein vitroandin vivo. Antigen-presenting cells incubated with this fusion protein were lysed by P18-I10-specific CTLin vitroand this activity was shown to be MHC class I restricted. The activity was inhibited by brefeldin A but was not inhibited by proteasome inhibitors, possibly because PT undergoes retrograde intracellular transport through the Golgi apparatus to the endoplasmic reticulum and delivers epitopes directly to nascent class I molecules. Mice immunized intraperitoneally with a single dose of the fusion protein without adjuvant raised a strong gp120-specific CTL response in the spleen. This CTL response was dependent on (1) the dose of fusion administered, (2) the fusion of the epitope with the toxin (since coadministration of peptide and toxin gave no response), and (3) the activity of CD8+cells. These data demonstrate that this detoxified derivative to PT, which is already a component of a licensed vaccine for humans, could represent a useful vaccine vector molecule for stimulation of HIV-specific CTL responses.