Role of Cryptosporidium Mucins in Host-parasite interactions

隐孢子虫粘蛋白在宿主-寄生虫相互作用中的作用

• 批准号: 7268032
• 负责人: ROBERTA M O'CONNOR
• 金额: $ 19.54万
• 依托单位: TUFTS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-06-15 至 2008-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7268032
• 关键词: Acquired Immunodeficiency Syndrome; Antibodies; Antigens; Apical; Applications Grants; Area; Biological Assay; Biological Process; Biology; CD4 Positive T Lymphocytes; CD7 gene; Carbohydrates; Cells; Chromosomes, Human, Pair 2; Complex; Cryptosporidiosis; Cryptosporidium; Cryptosporidium parvum; Data; Dependence; Development; Diarrhea; Disease; Disease Outbreaks; Doctor of Philosophy; Epithelial; Epithelial Cells; Epitope Mapping; Epitopes; Escherichia coli; Future; Gene Cluster; Gene Expression; Genes; Genome; Glycoproteins; Homologous Gene; Immune; Immune Sera; Immune response; Immunity; Immunocompromised Host; Immunofluorescence Immunologic; Individual; Infection; Infection prevention; Intervention; Intestines; Invaded; Investigation; Lectin; Life; Link; Localized; MHC antigen; Measures; Mediating; Medical; Molecular Weight; Monoclonal Antibodies; Mucins; Mus; Open Reading Frames; Parasites; Patients; Pattern; Personal Satisfaction; Play; Post-Translational Protein Processing; Process; Production; Propionibacterium acnes; Proteins; Recombinant Proteins; Recombinants; Relative (related person); Research Personnel; Resistance; Resolution; Reverse Transcriptase Polymerase Chain Reaction; Role; Role playing therapy; Sporozoites; System; T-Lymphocyte Epitopes; Testing; Therapeutic Corynebacterium Parvum; Time; Toxoplasma gondii; Vaccines; Virulent; Western Blotting; cell mediated immune response; cytokine; design; genome sequencing; glycosylation; insight; intracellular protein transport; mouse model; novel vaccines; pathogen; programs; protein expression; protein localization location; waterborne

DESCRIPTION (provided by applicant): Cryptosporidium is a ubiquitous waterborne pathogen that causes diarrheal disease worldwide. This pathogen is of significant medical importance because of its capacity to cause life-threatening diarrheal disease in immunocompromised hosts, particularly AIDS patients. To date there is no consistently effective treatment for Cryptosporidium infection, and no vaccine to prevent infection. Studies suggest that Cryptosporidium, unlike other apicomplexans, relies on mucin-like glycoproteins to attach to and invade intestinal epithelial cells. These mucin antigens are also targets of protective cellular immune responses. Despite the importance of mucin antigens to Cryptosporidium biology, few of these antigens have been investigated. The Cryptosporidium genome project has identified 31 open reading frames encoding putative mucin antigens. The studies proposed in this application will investigate 7 mucins that reside on a single locus on chromosome 2 (CpMuc1-7) and determine the role they play in host-parasite interactions. Aim #1: To characterize expression, localization, post-translational modifications, and potential multi-protein complex formation of the seven mucin-like glycoproteins of the CpMuc1-7 locus. We will quantify expression of the CpMucs during intracellular development using real-time RT-PCR, determine protein expression and localization, elucidate the glycosylation patterns of the native antigens, and determine if these antigens form multi-protein complexes. Aim #2: To determine if the CpMuc1-7 glycoproteins play a role in invasion and/or stimulation of cell mediated immune responses. We will determine if anti-CpMuc antibodies and recombinant proteins inhibit C. parvum invasion of intestinal epithelial cells, and determine if the CpMucs are activators of cellular immune responses that develop during infection in mice. In these studies, we will also explore the effect of glycosylation in host cell invasion and immune activation. Finally, employing a non-virulent T. gondii strain as an antigen delivery system, we will determine if immune responses targeted against one of the CpMucs provides protection from cryptosporidiosis. These studies will enhance our understanding of the role mucin-like glycoproteins play in the establishment and resolution of Cryptosporidium infections and greatly advance our ability to design effective treatments to combat this serious infectious threat around the world.

描述（由申请人提供）：隐孢子虫是一种普遍存在的水传播病原体，在世界范围内引起腹泻病。这种病原体具有重要的医学意义，因为它能够在免疫功能低下的宿主（尤其是艾滋病患者）中引起危及生命的腹泻疾病。迄今为止，还没有针对隐孢子虫感染的持续有效的治疗方法，也没有预防感染的疫苗。研究表明，与其他顶端复合物不同，隐孢子虫依赖粘蛋白样糖蛋白附着并侵入肠上皮细胞。这些粘蛋白抗原也是保护性细胞免疫反应的目标。尽管粘蛋白抗原对隐孢子虫生物学很重要，但对这些抗原的研究很少。隐孢子虫基因组计划已鉴定出 31 个编码假定粘蛋白抗原的开放阅读框。本申请中提出的研究将调查位于 2 号染色体上单个基因座 (CpMuc1-7) 上的 7 种粘蛋白，并确定它们在宿主-寄生虫相互作用中所发挥的作用。目标#1：表征 CpMuc1-7 基因座的七种粘蛋白样糖蛋白的表达、定位、翻译后修饰和潜在的多蛋白复合物形成。我们将使用实时 RT-PCR 定量细胞内发育过程中 CpMuc 的表达，确定蛋白质表达和定位，阐明天然抗原的糖基化模式，并确定这些抗原是否形成多蛋白复合物。目标#2：确定 CpMuc1-7 糖蛋白是否在细胞介导的免疫反应的侵袭和/或刺激中发挥作用。我们将确定抗 CpMuc 抗体和重组蛋白是否抑制微小念珠菌侵入肠上皮细胞，并确定 CpMuc 是否是小鼠感染过程中产生的细胞免疫反应的激活剂。在这些研究中，我们还将探讨糖基化对宿主细胞侵袭和免疫激活的影响。最后，采用无毒力的刚地弓形虫菌株作为抗原递送系统，我们将确定针对其中一种 CpMuc 的免疫反应是否可以提供针对隐孢子虫病的保护。这些研究将增强我们对粘蛋白样糖蛋白在隐孢子虫感染的建立和解决中所发挥的作用的理解，并极大地提高我们设计有效治疗方法以应对世界各地这一严重感染威胁的能力。

项目成果

Serum antibody responses to polymorphic Cryptosporidium mucin antigen in Bangladeshi children with cryptosporidiosis.

孟加拉国隐孢子虫病儿童对多态性隐孢子虫粘蛋白抗原的血清抗体反应。

• DOI: 10.4269/ajtmh.2011.11-0270
• 发表时间: 2011
• 期刊: The American journal of tropical medicine and hygiene
• 作者: Lai,Olivia;Morris,Christopher;Ahmed,Sabeena;Karim,MohammedMahbubul;Khan,Wasif;Ward,Honorine;O'Connor,Roberta
• 通讯作者: O'Connor,Roberta