NASAL DNA/PROTEIN VACCINE FOR ANTI-HIV ANTIBODY AND CTL

用于抗 HIV 抗体和 CTL 的鼻 DNA/蛋白疫苗

• 批准号: 6800281
• 负责人: PAMELA ANN KOZLOWSKI
• 金额: $ 53.63万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-03-01 至 2009-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6800281
• 关键词: AIDS vaccines; HIV envelope protein; Macaca mulatta; antigen antibody reaction; biotechnology; cellular immunity; dosage; enzyme linked immunosorbent assay; female; flow cytometry; human immunodeficiency virus; humoral immunity; immune response; immunization; immunomodulators; inhalation drug administration; interferon gamma; leukocyte activation /transformation; mucosa; mucosal immunity; recombinant proteins; simian immunodeficiency virus; subcutaneous drug administration; technology /technique development; vaccine development; vaccine evaluation; vector vaccine

DESCRIPTION (provided by applicant): This project will test the hypothesis that supplementation of a DNA vaccine with recombinant HIV envelope protein plus adjuvant will induce strong antiviral humoral as well as cellular immune responses in both mucosal and systemic compartments if simultaneously administered by both the intradermal and nasal routes. Specific Aim I is to confirm that the parenteral adjuvant QS-21 can also be used to adjuvant immune responses to nasally administered DNA vaccines or soluble protein, and to establish an effective nasal QS-21 dosage for these vaccine preparations. Groups of female rhesus macaques will be nasally vaccinated with 10 or 25 pg QS-21 plus rgp41-hemagglutinin fusion protein (gp41HA) or DNA encoding noninfectious SHIV89.6P particles (SHIV DNA). HIV- and SIV-specific antibodies induced in sera and secretions will be quantitated by ELISA. T helper cell (Th) and cytotoxic T lymphocyte (CTL) responses in the periphery, rectal mucosa, and the cervical/vaginal mucosa will be evaluated by testing isolated mononuclear cells for antigen-specific lymphoproliferation, IFN-( secretion, and intracellular IFN-gamma production using the [3H] thymidine uptake assay, ELISPOT assay, and flow cytometry. The QS-21 dose found to produce greatest immune responses would be used for subsequent nasal vaccinations. Specific AIM II is to use QS-21, SHIV DNA, and gp41HA in nonhuman primates to identify an optimal, practical DNA/protein vaccination protocol for induction of humoral and cellular immune responses in the systemic compartment, rectal mucosa, and genital tract mucosa. We will first confirm that SHIV DNA plus gp41HA and QS-21 can be administered as one formulation without loss of immunogenicity. Ultimately, a vaccination strategy involving simultaneous nasal/intradermal administration of SHIV DNA, gp41HA, and QS-21 followed by simultaneous nasal/intradermal boosting with recombinant modified vaccinia ankara virus-expressing SHIV antigens will be tested for immunogenicity, and protective efficacy after rectal SHIV89.6P challenge. In all of these studies, the methods described above in AIM I will be used to analyze antibody and T cell responses, including those in the mucosa. Specific Aim III is to refine and optimize an HIV/epithelial cell-binding assay that could be used to test secretions of vaccine recipients for antibodies that block HIV attachment to epithelial cells. Secretions collected from animals vaccinated above will be analyzed in this assay to determine if the gp41HA vaccine antigen is capable of inducing mucosal antibodies that could potentially prevent entry of HIV into the host. This study is expected to significantly further our knowledge concerning adjuvants for DNA vaccines as well as the nasal vaccination route, the immunogenicity of potential HIM vaccine subunit components, methods that could be used for evaluating secretions for transmission-preventing mucosal antibodies, and the design of vaccination protocols that are most likely to induce protective immunity to HIV.

描述(申请人提供)：该项目将测试这样一种假设，即如果同时通过皮内和鼻腔途径给药，补充含有重组HIV包膜蛋白和佐剂的DNA疫苗将在粘膜和全身两个部分诱导强烈的抗病毒体液和细胞免疫反应。 具体目的一是证实注射佐剂QS-21也可用于鼻用DNA疫苗或可溶性蛋白的佐剂免疫应答，并为这些疫苗制剂建立有效的鼻腔QS-21剂量。每组雌性猕猴将被鼻腔接种10或25pgQS-21加rgp41-血凝素融合蛋白(Gp41HA)或编码非传染性SHIV89.6P颗粒的DNA(SHIV DNA)。在血清和分泌物中诱导的HIV和SIV特异性抗体将通过ELISA法进行定量。外周、直肠粘膜和宫颈/阴道粘膜中的T辅助细胞(Th)和细胞毒性T淋巴细胞(CTL)的反应将通过检测分离的单个核细胞的抗原特异性淋巴增殖、干扰素分泌和细胞内干扰素-γ的产生来评估，方法是[~3H]胸腺嘧啶核苷摄取试验、ELISPOT试验和流式细胞术。被发现产生最大免疫反应的QS-21剂量将用于随后的鼻腔疫苗接种。 目的：利用QS-21、SHIV DNA和gp41HA在非人灵长类动物体内进行免疫接种，以确定一种最佳、实用的DNA/蛋白质免疫方案，以诱导体液免疫和细胞免疫应答。我们将首先确认SHV DNA加gp41HA和QS-21可以作为一个制剂使用，而不会失去免疫原性。最终，将测试一种疫苗接种策略，该策略包括同时鼻腔/皮内注射SHV DNA、gp41HA和QS-21，然后同时鼻腔/皮内接种表达SHIV病毒的重组改良痘苗病毒抗原，以检测免疫原性和直肠SHIV89.6P攻击后的保护效果。在所有这些研究中，上述AIM I中描述的方法将用于分析抗体和T细胞反应，包括粘膜中的抗体和T细胞反应。 具体目标三是改进和优化艾滋病毒/上皮细胞结合试验，该试验可用于测试疫苗接受者的分泌物中是否有阻止艾滋病毒附着于上皮细胞的抗体。从上述接种的动物身上收集的分泌物将在这项检测中进行分析，以确定gp41HA疫苗抗原是否能够诱导粘膜抗体，从而潜在地阻止艾滋病毒进入宿主。 这项研究有望极大地促进我们对DNA疫苗佐剂以及鼻腔接种途径的了解，潜在的HIM疫苗亚单位成分的免疫原性，可用于评估防止传播的粘膜抗体的方法，以及最有可能诱导对HIV的保护性免疫的疫苗接种方案的设计。