IMMUNOCONJUGATES FOR BOOSTING IMMUNITY IN HIV INFECTED PERSONS

HIV-1感染的早期表现是辅助T细胞的丧失 (CD4+)细胞功能。随后，这些细胞的数量也被耗尽 提示HIV-1的细胞病变效应。由于抗体中需要CD4+ 对蛋白质抗原的反应，功能或数量上的缺陷 预计细胞会导致抗体反应受损。 HIV-1。我们的方法是开发一种疫苗来增强人类的免疫反应 HIV-1感染者一直使用T细胞非依赖性(TI)抗原 从而绕过了对CD4+T细胞的要求。众所周知，布鲁氏菌 流产(BA)可作为半抗原的载体刺激抗体 在没有T细胞的情况下的反应。之前我们已经证明了HIV-1 与流产布鲁氏菌(BA)结合可诱导抗HIV抗体应答 即使在耗尽了CD 4+T细胞的小鼠中也是如此。由于脂多糖(LPS)来自于 其他革兰氏阴性杆菌也能刺激B细胞，我们决定提纯内毒素 来自BA，如果被激活，可以取代BA作为载体。这将提炼出 使用BA作为载体，使分子具有确定的结构并消除 其他可能有毒的BA元素。用正丁醇提纯BA中的脂多糖 并用KDO、蛋白质和核酸含量进行表征。它 也进行了功能活动测试。LP-BA表现为与T无关 I型携带者，因为(I)它能有效刺激小鼠脾细胞 即使在T细胞耗尽后仍能增殖并分泌抗体；以及 (2)TNP-内毒素(BA)可诱导各亚类抗TNP抗体。

An early manifestation of infection with HIV-1 is the loss of helper T (CD4+) cell function. Subsequently these cells are also depleted in number cue to cytopathic effects of HIV-1. Since CD4+ are required in antibody responses to protein antigens, deficiency in function or number of these cells would be expected to result in impaired antibody responses against HIV-1. Our approach for developing a vaccine to boost immune responses in HIV-1 infected individuals has been to use T-cell independent (TI) antigens and thus bypass he requirement for CD4+ T cells. It is known that Brucella abortus (BA) can be used a carrier for haptens to stimulate antibody responses in the absence of T cells. Previously we showed that HIV-1 conjugated to Brucella abortus (BA) elicited anti-HIV antibody responses even in mice depleted od CD4+ T cells. Since lipopolysaccharide (LPS) from other gram negative bacilli can stimulate B cells, we decided to purify LPS from BA which, if active could replace BA as the carrier. This would refine the use of BA as a carrier to a molecule of defined structure and eliminate other BA elements that may be toxic. LPS from BA was purified by butanol extraction and characterized by KDO, protein and nucleic acid content. It was also tested for functional activity. LPS-BA behaved as T-independent type I carrier since (i) it was effective in stimulating mouse spleen cells to proliferate and to secrete antibody even after T cell depletion; and (ii) TNP-LPS (BA) could induce anti-TNP antibodies of all IgG subclasses.