MUCOSAL IMMUNE RESPONSE TO HIV GP120 VACCINES

这个粘液免疫学协作组的目标是提供 基本信息，将导致粘膜疫苗的设计， 在直肠和宫颈中引发抗gp 120分泌型伊加（sIgA）抗体 女性的分泌物，并减少性或垂直的风险， 艾滋病的传播。项目1的重点是增强M细胞对 口服或直肠施用的含gp 120的疫苗， 直肠和女性生殖器粘膜表面上的抗gp 120伊加抗体， 并测试特定IgA的保护潜力。该项目 三个目标： 目的I是测试靶向gpl 20蛋白或 肽在肠和直肠诱导位点对M细胞的作用，以及 评价直肠、子宫颈和阴道中产生的抗HIV sIgA。选择 重组gp 120和多聚gp 120肽（包括霍乱毒素 由项目2提供的缀合物）将在靶向M细胞的细胞上递送。 通过口服和直肠给药的颗粒载体和脂质体，和 通过直接应用于卵泡相关上皮细胞。替代 将通过定量M细胞粘附来评价策略， 运输，并通过ELISA测量分泌物中的特异性伊加水平 用吸收芯从局部粘膜表面取回。 目的II是确定是否活的减毒SIV颗粒是 小鼠直肠或肠M细胞转运效率， 猴，并评估局部粘膜免疫反应，这种类型的 候选疫苗。 将三重减毒SIV应用于 体外猴直肠粘膜，以及小鼠/兔Peyer集合淋巴结和直肠 体内粘膜。M细胞转运将通过EM和免疫- 细胞化学猴和小鼠将通过直肠或阴道免疫 减毒SIV和局部（直肠和宫颈）抗SIV伊加 分泌将通过ELISA/芯吸法测量。 目的III是制备和表征抗gp 120单克隆伊加 识别并沉淀完整HIV或SIV颗粒的抗体， 并测试它们防止病毒接触上皮细胞的能力， 靶细胞表面。使用有效的微粒疫苗， 目的I和来自目的II的减毒SIV，将粘膜免疫小鼠， 伊加杂交瘤将从派伊尔集合淋巴结或直肠淋巴结产生。 淋巴细胞的表位特异性，以及抗gp 120单克隆抗体的表位特异性。 将确定IgA。将测试IgA的能力， 沉淀HIV，在体外保护HIV易感细胞， 防止病毒、gp 120包被的颗粒或gp 120表达的颗粒接触 细胞与上皮表面在体外。 在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，并将有助于设计 一种有效的抗艾滋病粘膜疫苗

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.