IMMUNIZATION AGAINST LENTIVIRAL GP120 USING SV40 VECTORS

描述：(改编自申请者摘要)有效、安全的目标 针对HIV-1的疫苗尚未实现。尽管有希望的概念 已经进行了检测，对HIV-1免疫生物学有了相当大的了解， 艾滋病毒-1感染和艾滋病的有效预防仍然难以捉摸。在此R21中 应用，我们建议测试一种新型的基因递送载体，重组 猴病毒-40(RSV40)作为传递HIV-1和SIV gp120信封的载体 糖蛋白。RSV40作为这种类型的载体有几个潜在的优势 关于免疫的问题。它能感染大多数细胞类型，在冻干状态下很稳定， 即使在室温下，也可以在非常高的滴度下生产。同样，rSV40 不会引起任何可检测到的中和抗体，因此可以给出 多次，以增强或增强对(例如)的免疫反应虚弱的 免疫原性转基因。我们假设携带慢病毒gp120的rSV40可以 同时诱导大量抗体和细胞毒性T淋巴细胞(CTL)反应 抗gp120。我们已经构建了携带HIV-INL4-3 gp120的rSV40 基因(SVgp120)，表达gp120。BALB/c小鼠接种繁殖体 二次接种后，SVgPL20产生大量的抗gp120抗体。 在此应用程序中，我们提出了Jefferson与 医学院和新英格兰地区灵长类研究中心进行测试 RSV40诱导免疫应答的能力与gpl20相比，并评估 保护您免受SIV的挑战。因此，我们将给BALB/c小鼠接种 SVgpl20每月，使用携带NL4-3的牛痘病毒进行或不使用预接种 环境将测定与gp20的中和和结合抗体及CTL。这个 然后将在恒河猴身上测试最有效的免疫方案。 猴子。我们将使用SIVmac239 gp130(SVgpl30)制作rSV40。应用 SVgpl20免疫数据，我们将SVgpl30接种BALB/c，测量 抗SIVmac239 gpl30的抗体和CTL，然后在恒河猴中重复这些研究 猕猴。然后，免疫的猴子将受到致病病毒的攻击 SIVmac239，并评估疾病进展。此方法将测试 慢病毒rSV40载体多次接种的原理 GPL20可以增强对这些弱抗原的体液和细胞免疫，并且 提供对免疫抑制慢病毒感染的保护。

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.