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

HIV-1感染的特征是功能下降， 辅助功能所需的CD4 + T细胞数量 对蛋白质抗原的抗体反应。 我们之前已经证明， 小鼠中对HIV-1蛋白的抗体应答是T辅助细胞依赖性的， 而HIV蛋白质和肽，与流产布鲁氏菌（BA）结合， 不需要CD4 + T细胞。 随后，我们将18个氨基 来源于HIV-MN的第三可变区的酸性肽（N3V3） 包膜，含有B和CTL表位，对BA（N3 V3-BA）和对 匙孔血蓝蛋白（N3V3-KLH）。 两种偶联物均具有免疫原性 在正常小鼠中。 IgG 1是V3-KLH诱导的主要同种型， 而IgG 2a是N3V3-BA诱导的优势IgG亚类。 CTL 也引发，由任何构建体，这可以裂解肽脉冲 和HIV-1感染的靶细胞。 由于BA作为载体可以在CD4+耗竭的小鼠中诱导抗体， 由于BA可以从CD8 + T细胞中诱导细胞因子，我们 研究了是否可以在缺乏CTL和抗体的小鼠中诱导CTL和抗体， CD4 + T细胞。 N_3V_3-BA能诱导抗体和CTL的产生，而N_3V_3-KLH不能 小鼠的CD4 + T细胞耗竭。 此外，来自 测试用N3V3-缀合物免疫的小鼠 中和不同的HIV-1分离株分离株保留 G-P-G-R-A-F，即IIIB、MN和SF 2被中和（> 50%合胞体 抑制），滴度为160或 更大 然而，该部位的射频隔离物不同 （G-P-G-R-V-I）以较低滴度中和。 这些结果表明 那个B。在亚单位的发育过程中， 设计用于产生抗HIV抗体和CTL应答的疫苗， CD4 + T细胞功能受损的人。

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.