IMMUNIZATION AGAINST LENTIVIRAL GP120 USING SV40 VECTORS

描述：（改编自申请人的摘要）有效、安全、 针对HIV-1的疫苗尚未实现。虽然有前途的概念 已经进行了测试，并获得了对HIV-1免疫生物学的相当多的了解， HIV-1感染和AIDS的有效预防仍然是难以捉摸的。在R21中 应用，我们建议测试一种新的基因传递载体，重组 猴病毒-40（rSV40），作为传递HIV-1和SIV gp120包膜载体 糖蛋白rSV 40作为该类型的载体具有几个潜在优势 免疫。它感染大多数细胞类型，在冻干形式下是稳定的， 甚至在室温下也可以制备，并且可以以非常高的滴度制备。另外，rSV40 不会引起任何可检测的中和抗体，因此可以给予 多次，以加强或增强针对（例如）弱 免疫原性转基因。我们假设携带慢病毒gp120的rSV 40可以 引发大量抗体和细胞毒性T淋巴细胞（CTL）应答 针对GP120。我们构建了携带HIV-1 NL4 - 3 gp120的rSV 40， 基因（SVgp120）并表达gp120。BALB/c小鼠多次接种 SVgp120通过第二次接种产生针对gp120的实质性抗体。 在这个应用程序中，我们提出了一个合作项目之间的杰斐逊 医学院和新英格兰地区灵长类动物研究中心进行测试 rSV 40相对于gp120引发免疫应答的能力，以及评估 保护免受SIV的挑战。因此，我们将BALB/c小鼠与 SVgp 120，每月一次，使用携带NL4 - 3的牛痘病毒进行或不进行引发 env.将测量中和和结合抗体以及针对gpl20的CTL。的 然后在恒河猴中测试最有效的免疫方案 猴子我们将用SIVmac239 gpl30（SVgpl30）制备rSV 40。应用 SVgpl20免疫数据，我们将用SVgpl30接种BALB/c，测量 然后在恒河猴中重复这些研究 猕猴免疫的猴子将被攻击致病性 SIVmac239并评估疾病进展。这种方法将测试 用携带慢病毒的rSV 40载体多次接种的原理 gp120可以增强对这些弱抗原的体液和细胞免疫， 提供免疫抑制性慢病毒感染的保护。

DESCRIPTION: (Adapted from Applicant's Abstract) The goal of an effective, safe vaccine against HIV-1 has not yet been realized. Although promising concepts have been tested and considerable understanding of HIV-1 immunobiology gained, effective prophylaxis of HIV-1 infection and AIDS remains elusive. In this R21 application, we propose to test a novel gene delivery vehicle, recombinant simian virus-40 (rSV40), as a vehicle to deliver HIV-1 and SIV gp120 envelope glycoproteins. rSV40 has several potential strengths as a vector for this type of immunization. It infects most cell types, is stable in lyophilized form, even at room temperature, and can be made at very high titer. As well, rSV40 does not elicit any detectable neutralizing antibodies, and so may be given multiple times, to boost or enhance immune responses against (e.g.) weakly immunogenic transgenes. We hypothesize that rSV40 carrying lentivirus gp120 can elicit both substantial antibody and cytotoxic T lymphocyte (CTL) responses against gp120. We have constructed a rSV40 that carries the HIV- INL4-3 gp120 gene (SVgp120) and expresses gp120. BALB/c mice inoculated multiply with SVgpl20 produce substantial antibody against gp120 by the second inoculation. In this application, we propose a collaborative project between Jefferson Medical College and the New England Regional Primate Research Center to test the ability of rSV40 to elicit immune responses vs. gpl20, and to evaluate protection from challenge with SIV. Thus, we will inoculate BALB/c mice with SVgpl20 monthly, with or without priming using vaccinia virus carrying NL4-3 env. Neutralizing and binding antibodies and CTL to gpl20 will be measured. The most effective immunization regimen will then be tested in rhesus macaque monkeys. We will make a rSV40 with SIVmac239 gp130 (SVgpl30). Applying the SVgpl20 immunization data, we will inoculate BALB/c with SVgpl30, measuring antibody and CTL to SIVmac239 gpl30, then repeat these studies in rhesus macaque monkeys. Immunized monkeys will then be challenged with pathogenic SIVmac239 and evaluated for disease progression. This approach will test the principle that multiple inoculations with a rSV40 vector bearing lentiviral gpl20 can enhance humoral and cellular immunity to these weak antigens, and afford protection from immunosuppressive lentivirus infection.