Mimotope Conversion of HIV-1 Carbohydrate Antigens

碳水化合物抗原一直是激发对许多病原体的保护性免疫反应的基础，但这种方法在艾滋病毒研究中还没有得到充分的评估。抗碳水化合物抗体可能有助于抑制感染细胞与粘膜树突状细胞的结合，从而阻止早期感染。我们已经开发了一个项目，涉及将与HIV相关的碳水化合物表位相互转化为基于多肽的疫苗策略，以增强碳水化合物的交叉反应，全身和粘膜反应。在特定的目标1中，我们检验了HIV-1相关碳水化合物抗原的多肽模拟表位诱导的免疫在定性和定量上不同于碳水化合物抗原本身诱导的免疫的假设。有趣的是多克隆反应的评价。抗体诱导的动力学。碳水化合物/gp120反应性同工型图谱；糖类/gp120反应性抗体亲和力/亲和力。评估血清与暴露在天然形式的gp120和gp160上的表位的结合，以及单体gp120和细胞表面表达的寡聚体gp120/gp41的异源菌株。在具体目标2中，我们探索了提高碳水化合物反应滴度和扩大艾滋病毒免疫反应广度的方法。在用模拟或碳水化合物配方启动并用低聚-gp160配方增强后，评估全身和粘膜反应。血清和粘膜灌洗液用于I。实验室和初级分离株的中和试验II。抑制树突状细胞黏附和感染3。还评估了与目标1中的gp120表位结合的血清抗体。在具体目标3中，利用分子模拟和噬菌体展示文库的系统方法来分析和利用拓扑相似性来进一步确定HIV-1相关糖类抗原的原型多肽模拟表位模板。在这一目标中，HIV-1相关甘露糖醇和乳糖系列碳水化合物结构之间的拓扑关系与与选定凝集素反应的多肽模拟表位和高效碳水化合物反应性HIV-1中和抗体2G12的结合特性相关。我们进一步定义了与这些受体高亲和力结合的多肽的变体。)合成和生物扫描定义的代表反映碳水化合物和多肽结构的二级结构元素的多肽阵列文库；以及2)通过Biacore和计算机模拟研究评估配体结合的亲和力。

Carbohydrate antigens have been the basis for eliciting protective immune responses against many pathogens, yet this approach has not been adequately assessed in HIV research. Anti- carbohydrate antibodies may facilitate the inhibition of infected cell binding to dendritic cells on mucosa to thwart early infection. We have developed a program concerned with the inter- conversion of carbohydrate epitopes associated with HIV into a peptide based vaccine strategy to augment carbohydrate cross- reactive systemic and mucosal responses. In specific aim 1, we test the hypothesis that immunity induced by peptide mimeotopes of HIV-1 associated carbohydrate antigens differs qualitatively and quantitatively from the immunity induced by the carbohydrate antigen itself. Of interest is the evaluation of polyclonal responses in terms of i. kinetics of antibody induction; ii. Carbohydrate/gp120 reactive isotype profiles; and iii. Carbohydrate/gp120 reactive antibody affinity/avidity. Evaluation of serum binding to epitopes exposed on native forms of gp120 and gp160, and heterologous strains of monomeric gp120 and cell surface-expressed oligomeric gp120/gp41 are evaluated. In specific aim 2, we explore ways to enhance carbohydrate reactive titers and expand on the breadth of HIV immunoreactivity. Systemic and mucosal responses are evaluated following priming with mimetic or carbohydrate formulations and boosting with oligomeric-gp160 formulates. Serum and mucosal lavages are tested for i. Neutralization of Lab and primary isolates ii. Inhibition of Dendritic cell adherence and infection iii. Evaluation of serum antibody binding to gp120 epitopes as in aim 1 are also evaluated. In specific aim 3, systematic approaches involving molecular modeling and phage display libraries are used to analyze and exploit topological similarities to further define prototypic peptide mimeotope templates of HIV-1 associated carbohydrate antigens. In this aim, topological relationships between HIV-1 associated mannosyl and lactoseries carbohydrate structures are correlated with binding properties of peptide mimeotopes reactive with selected lectins and a highly effective carbohydrate reactive HIV-1 neutralizing antibody 2G12. We further define variants of peptides that display high affinity binding to these receptors by 1.) Synthesizing and biopanning defined peptide array libraries representative of secondary structure elements reflective of carbohydrate and peptide structures; and 2.)Evaluation of the affinity of ligand binding by Biacore and computer modeling studies.