DEVELOPMENT OF T-CELL INDEPENDENT VACCINES FOR HIV-1 AND OTHER INFECTIOUS AGENTS

感染HIV-1的特征是功能下降和 辅助性功能所需的CD4+T细胞数量 对蛋白质抗原的抗体反应。我们之前已经表明， 小鼠对HIV-1蛋白的抗体反应依赖于T辅助细胞， 而与布鲁氏菌结合的HIV蛋白和多肽(BA) 不需要CD4+T细胞。随后，我们偶联了18个氨基 HIV-MN第三可变区的酸性多肽(N3V3) 包膜，包含B和CTL表位，到BA(N3V3-BA)和TO 锁孔帽状血蓝蛋白(N3V3-KLH)。这两种结合物都具有免疫原性。 在正常小鼠身上。V3-KLH诱导的同工型以IgG1为主， 而IgG2a是N3V3-BA诱导的主要免疫球蛋白亚类。CTL 也可以通过这两种结构中的任何一种来诱导，这种结构可以裂解多肽脉冲 和HIV-1感染的靶细胞以MHC I类限制性方式。 由于BA作为载体可以在CD4+耗尽的小鼠中诱导抗体 因为BA可以从CD8+T细胞中诱导细胞因子，所以我们 研究是否可以在缺乏CTL和抗体的小鼠中诱导CTL和抗体 CD4+T细胞。N3V3-BA，而不是N3V3-KLH，既能引起抗体又能引起CTL CD_4~+T细胞耗尽小鼠的反应。此外，来自俄罗斯的血清 用N3V3结合物免疫的小鼠进行了能力测试。 中和不同的HIV-1分离株。保留病毒的分离株 G-P-G-R-A-F，即IIIB、MN和SF2被中和(&gt；50%合胞体 效价160或以上的N3V3-BA免疫小鼠血清 更大。然而，射频隔离在这个站点是不同的 (G-P-G-R-V-I)中和效价较低。这些结果表明 该菌可作为亚基发育的载体 旨在产生抗HIV抗体和CTL反应的疫苗 CD4+T细胞功能受损的个体。

Infection with HIV-1 is characterized by a decrease on function and number of CD4+ T cells which are required for helper function in the antibody response to protein antigens. We have previously shown that antibody responses to HIV-1 proteins in mice are T-helper cell dependent, whereas HIV proteins and peptides, conjugated to Brucella abortus (BA) do not require CD4+ T cells. Subsequently, we conjugated an 18 amino acid peptide (N3V3) derived from the third variable region of HIV-MN envelope, containing both B and CTL epitopes, to BA (N3V3-BA) and to keyhole limpet hemocyanin (N3V3-KLH). Both conjugates were immunogenic in normal mice. IgG1 was the predominant isotype induced by V3-KLH, whereas IgG2a was the preeminent IgG subclass elicited by N3V3-BA. CTL were also elicited, by either construct, which could lyse peptide-pulsed and HIV-1-infected target cells in an MHC class I restricted manner. Since BA, as a carrier, can induce antibodies in mice depleted of CD4+ T cells and since BA can induce cytokines from CD8+ T cells, we investigated whether CTL and antibodies could be induced in mice lacking CD4+ T cells. N3V3-BA, but not N3V3-KLH, evoked both antibody and CTL responses in mice depleted of CD4+ T cells. In addition, sera from the mice immunized with the N3V3-conjugates were tested for ability to neutralize different HIV-1 isolates. Isolates which retain the G-P-G-R-A-F, namely, IIIB, MN and SF2 were neutralized (>50% syncytia inhibition) by sera from N3V3-BA immunized mice at titers of 160 or greater. The RF isolate, however, which is different at this site (G-P-G-R-V-I) was neutralized at a lower titer. These results indicate that B. abortus can be used as a carrier in the development of subunit vaccines designed to generate anti-HIV antibody and CTL responses in individuals with impaired CD4+ T cell function.