IMMUNOBIOLOGY OF A 30 KDA PROTEIN OF HAEMOPHILUS DUCREYI

杜克雷嗜血杆菌 30 KDA 蛋白的免疫生物学

• 批准号: 6347207
• 负责人: CHRISTOPHER ELKINS
• 金额: $ 15.71万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-01 至 2001-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6347207
• 关键词: Haemophilus; adhesin; bacteria infection mechanism; bacterial antigens; bacterial proteins; bacterial vaccines; clinical research; cooperative study; epithelium; host organism interaction; human subject; laboratory rabbit; molecular cloning; neutrophil; protein structure function; recombinant proteins; sexually transmitted diseases; vaccine development; virulence; Haemophilus; adhesin; bacteria infection mechanism; bacterial antigens; bacterial proteins; bacterial vaccines; clinical research; cooperative study; epithelium; host organism interaction; human subject; laboratory rabbit; molecular cloning; neutrophil; protein structure function; recombinant proteins; sexually transmitted diseases; vaccine development; virulence

This project seeks to understand the molecular interactions of Haemophilus decreyi with human neutrophils (PMN) and host epithelial cells during the initial stages of infection. Attachment to, and internalization of, pathogens by host cells is often the first step in the infectious process. In vitro, H. ducreyi is bound and internalized by PMN in the absence of serum opsonins and by epithelial cells suggesting a ligand (adhesin)/receptor interaction(s). Immunological intervention early in the infectious process could modify or prevent cellular attachment and possibly disease. PMNs are the first cells recruited at the onset of chancroidal lesion and therefore might be an important defense against infection. These studies will enhance our overall understanding of the role of attachment to epithelial cells and PMN in the infectious process, the ability of PMN to kill H. ducreyi, and perhaps, lead to new vaccine strategies. There are five Specific Aims. In the first Specific Aim, an isogenic mutant of the 30 kDa outer membrane protein will be constructed. As described in detail in the following sections, the 30 kDa protein is related to cellular adhesins and proteins which mediate serum resistance in other bacteria. The second Specific aim is to determine the function of this protein. We will use mutant/parent comparisons to determine its possible role in adherence to PMN, epithelial cells, and cellular matrix proteins,. We will also perform 30 kDa mutant/parent comparisons for resistance to serum and phagocytic killing. In the third Specific aim, we will test an isogenic mutant of the H. ducreyi 30 kDa protein in the temperature dependent rabbit model of infection and in the human experimental model of H. ducreyi infection. The fourth Specific aim addresses certain vaccine related issues. We will determine if antibodies to the 30 kDa protein are elicited during chancroid infection to document in vivo expression. We will test the ability of purified recombinantly expressed 30 kDa to protect animals from a challenge infection. In Specific Aim 5 we will identify PMN and cellular adhesins other than the 30 kDa protein.

该项目旨在理解Decreyi与人类嗜中性粒细胞（PMN）和宿主上皮细胞的分子相互作用。宿主细胞对病原体的附着和内在化通常是传染过程中的第一步。在体外，在没有血清opsonins和上皮细胞的情况下，杜克里氏菌由PMN结合并内化，表明配体（粘附素）/受体相互作用（S）。在传染过程的早期，免疫学干预可以改变或防止细胞附着和可能的疾病。 PMN是在结构病变开始时招募的第一个细胞，因此可能是针对感染的重要防御。这些研究将增强我们对上皮细胞和PMN的作用的总体理解，在传染过程中，PMN杀死H. ducreyi的能力，也许会导致新的疫苗策略。有五个具体目标。在第一个特定的目的中，将构建30 kDa外膜蛋白的同基因突变体。如以下各节中详细描述的，30 kDa蛋白与细胞粘附素和蛋白质有关，这些蛋白和蛋白质介导其他细菌中的血清耐药性。第二个特定目的是确定该蛋白质的功能。我们将使用突变/父对比较来确定其在遵守PMN，上皮细胞和细胞基质蛋白中的可能作用。我们还将进行30 kDa突变体/父母的比较，以抗血清和吞噬细胞杀伤。在第三个特定目的中，我们将在温度依赖性的感染兔模型和H. ducreyi感染的人类实验模型中测试30 kDa蛋白的同基因突变体。第四个特定目的解决了某些与疫苗相关的问题。我们将确定在牙冠感染期间是否会引起30 kDa蛋白的抗体以记录体内表达。我们将测试纯化的重组表达30 kDa以保护动物免受挑战感染的能力。 在特定目标5中，我们将鉴定出30 kDa蛋白以外的PMN和细胞粘附素。