Laube L S, Burrascano M, Dejesus C E, Howard B D, Johnson M A, Lee W T, Lynn A E, Peters G, Ronlov G S, Townsend K S
Viagene, Inc., San Diego, CA 92121.
Hum Gene Ther. 1994 Jul;5(7):853-62. doi: 10.1089/hum.1994.5.7-853.
The immune response against human immunodeficiency virus type-1 (HIV-1) is believed to play a role in controlling the early stages of disease progression. The cellular immune response, in particular cytotoxic T lymphocyte (CTL) activity, may be important for eliminating virally infected cells in HIV-1-infected individuals. Genetic immunization using retroviral vectors provides an effective means of introducing antigens into the antigen presentation pathways for T cell stimulation. A nonreplicating, amphotropic murine retroviral vector containing the HIV-1 IIIB env gene has been used to transduce primary rhesus monkey fibroblasts for the expression of HIV-1 antigenic determinants. Rhesus monkeys were immunized with four doses of either vector-transduced autologous fibroblasts (VTAF) expressing the HIV-1 IIIB ENV/REV proteins or nontransduced autologous fibroblasts (NTAF) administered at 2-week intervals. The animals were evaluated for both the induction of HIV-1-specific immune responses and potential toxicity associated with this ex vivo treatment. The VTAF-immunized monkeys generated CTL responses specific for HIV-1 ENV/REV expressing autologous target cells, whereas, NTAF-immunized monkeys showed negligible CTL activity. The cytotoxic activity was mediated by CD8+, major histocompatibility complex (MHC)-restricted CTL. In addition, antibody responses directed against the HIV-1 gp120 protein were also detected in the sera of VTAF-immunized monkeys. Clinical and histopathological evaluation of immunized monkeys showed no evidence of significant adverse events. Several animals that received either VTAF or NTAF had detectable anti-cytoplasmic antibodies, but were not positive for anti-nuclear antibodies or rheumatoid factor. Subsequent evaluation of renal, synovial, and hepatic tissue samples from these monkeys revealed no autoimmune disease-associated lesions. This study demonstrates the safety and ability of autologous retroviral vector-transduced cells expressing HIV-1 IIIB ENV/REV proteins to stimulate immune responses in a non-human primate model, and provides a basis for this form of genetic immunization in HIV-infected humans.
针对1型人类免疫缺陷病毒（HIV-1）的免疫反应被认为在控制疾病进展的早期阶段发挥作用。细胞免疫反应，特别是细胞毒性T淋巴细胞（CTL）活性，对于清除HIV-1感染个体中被病毒感染的细胞可能很重要。使用逆转录病毒载体进行基因免疫提供了一种将抗原引入抗原呈递途径以刺激T细胞的有效方法。一种含有HIV-1 IIIB env基因的非复制型、嗜性鼠逆转录病毒载体已被用于转导原代恒河猴成纤维细胞，以表达HIV-1抗原决定簇。恒河猴每隔2周接受四剂表达HIV-1 IIIB ENV/REV蛋白的载体转导自体成纤维细胞（VTAF）或未转导自体成纤维细胞（NTAF）免疫。对动物进行HIV-1特异性免疫反应的诱导以及与这种体外治疗相关的潜在毒性评估。VTAF免疫的猴子产生了针对表达HIV-1 ENV/REV的自体靶细胞的CTL反应，而NTAF免疫的猴子显示出可忽略不计的CTL活性。细胞毒性活性由CD8 +、主要组织相容性复合体（MHC）限制的CTL介导。此外，在VTAF免疫的猴子血清中也检测到了针对HIV-1 gp120蛋白的抗体反应。对免疫猴子的临床和组织病理学评估未发现明显不良事件的证据。接受VTAF或NTAF的几只动物检测到抗细胞质抗体，但抗核抗体或类风湿因子呈阴性。随后对这些猴子的肾脏、滑膜和肝脏组织样本进行评估，未发现自身免疫性疾病相关病变。这项研究证明了表达HIV-1 IIIB ENV/REV蛋白的自体逆转录病毒载体转导细胞在非人类灵长类动物模型中刺激免疫反应的安全性和能力，并为HIV感染人类的这种基因免疫形式提供了基础。
