HIV/SIV--MECHANISMS OF MUCOSAL TRANSMISSION & PROTECTION

该合作粘膜免疫学小组的目标是提供 将导致设计粘膜疫苗的基本信息 直肠和宫颈组织中抗gpl2O分泌型IgA抗体的诱导 女性分泌物，降低性行为或性行为的风险 艾滋病的传播。L项目的重点是增强M细胞对 口服或直肠注射含有gpl2O的疫苗，诱导 直肠和女性生殖器粘膜表面的抗gpl2O IgA抗体， 以及测试特定IGAs的保护潜力。该项目已 三个目标： 目的I是测试靶向gpl2O蛋白或 多肽与肠道和直肠诱导部位的M细胞，以及 评估在直肠、宫颈和阴道中产生的抗HIV SLGA。已选择 重组gpl2O和聚合gpl2O多肽(包括霍乱毒素 由项目2提供的结合物)将以M细胞为目标进行交付 口服和直肠给药的微粒载体和脂质体，以及 通过直接应用于毛囊相关的上皮细胞。备择 这些策略将通过量化M细胞黏附和 运输，并用酶联免疫吸附试验测定分泌物中的特异性IgA水平 用吸水灯芯从局部粘膜表面取回。 目的二是确定活的、衰减的SIV颗粒是否 小鼠直肠或肠道M细胞转运效率和 并评估局部粘膜对这种类型的免疫反应。 候选疫苗。三倍减毒SIV将应用于黄瓜外植体 猴直肠黏膜的体外培养及小鼠/兔Peyer‘s补片和直肠 活体内的粘膜。M细胞转运将通过EM和免疫学方法进行评估。 细胞化学。将对猴子和小鼠进行直肠或阴道免疫 使用减毒的SIV和局部(直肠和宫颈)抗SIV IgA 分泌物将用酶联免疫吸附试验/灯芯法测定。 目的III制备抗gpl2O单抗IgA并对其进行鉴定 识别并沉淀完整的HIV或SIV颗粒的抗体， 并测试它们防止病毒接触上皮细胞和 目标细胞表面。使用有效的颗粒疫苗 AIM I和来自AIM II的减毒SIV，对小鼠进行粘膜免疫， IgA杂交瘤将由Peyer‘s补片或直肠产生 淋巴细胞和抗gpl2O单抗的表位特异性 将确定政府间气候变化框架。将对IGAS进行测试，以确定它们的能力 使艾滋病毒沉淀，在体外保护艾滋病毒敏感细胞，并 防止病毒、包裹gpl2O的颗粒或表达gpl2O的颗粒接触。 体外培养有上皮面的细胞。 从这个项目中获得的信息将澄清潜在的作用 粘膜免疫在预防艾滋病中的作用，并将有助于设计 一种有效的针对艾滋病毒的粘膜疫苗。

The goal of this Collaborative Mucosal Immunology Group is to provide basic information that will lead to design of mucosal vaccines that elicit anti-gpl2O secretory IgA (slgA) antibodies in rectal and cervical secretions of women, and that reduce the risk of sexual or vertical transmission of AIDS. Project l is focused on enhancing M cell uptake of gpl2O-containing vaccines administered orally or rectally, induction of anti-gpl2O IgA antibodies on rectal and female genital mucosal surfaces, and testing the protective potential of specific IgAs. The project has three aims: Aim I is to test novel methods for targeting of gpl2O proteins or peptides to M cells at intestinal and rectal inductive sites, and to evaluate resulting anti-HIV slgA in rectum, cervix and vagina. Selected recombinant gpl2O and polymeric gpl2O peptides (including cholera toxin conjugates provided by Project 2) will be delivered on M cell-targeted particulate carriers and liposomes by oral and rectal administration, and by direct application to follicle-associated epithelia. Alternative strategies will be evaluated by quantitating M cell adherence and transport, and by ELISA measurement of specific IgA levels in secretions retrieved with absorbent wicks from local mucosal surfaces. Aim II is to determine whether live, attenuated SIV particles are efficiency transported by rectal or intestinal M cells in mice and monkeys, and to assess the local mucosal immune response to this type of candidate vaccine. Triple-attenuated SIV will be applied to explants of monkey rectal mucosa in vitro, and mouse/rabbit Peyer's patch and rectal mucosa in vivo. M cell transport will be assessed by EM and immuno- cytochemistry. Monkeys and mice will be immunized rectally or vaginally with attenuated SIV, and local (rectal and cervical) anti-SIV IgA secretion will be measured by the ELISA/wick method. Aim III is to produce and characterize anti-gpl2O monoclonal IgA antibodies that recognize and precipitate intact HIV or SIV particles, and to test their abilities to prevent viral contact with epithelial and target cell surfaces. Using effective particulate vaccines identified in Aim I and attenuated SIV from Aim II, mice will be mucosally-immunized, IgA hybridomas will be generated from Peyer's patch or rectal lymphocytes, and the epitope specificities of the anti-gpl2O monoclonal IgAs will be determined. IgAs will be tested for their capacity to precipitate HIV, to protect HIV-susceptible cells in vitro, and to prevent contact of virus, gpl2O-coated particles, or gpl2O-expressing cells with epithelial surfaces in vitro. The information gained in this project will clarify the potential role of mucosal immunity in prevention of AIDS, and will contribute to design of an effective mucosal vaccine against HIV.