VERMONT COBRE: PROJECT 2: INNATE IMMUNE RESPONSES TO CRYPTOSPORIDIUM PARVUM

佛蒙特州 COBRE：项目 2：对隐孢子虫的先天免疫反应

• 批准号: 8167731
• 负责人: Beth Diane Kirkpatrick
• 金额: $ 16.86万
• 依托单位: UNIVERSITY OF VERMONT & ST AGRIC COLLEGE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8167731
• 关键词: Acquired Immunodeficiency Syndrome; Antibodies; Attenuated; Bangladesh; Biological Assay; Birth; Cell Line; Cellular Assay; Cellular Immunology; Centers of Research Excellence; Child; Clinical Data; Clinical Research; Collectins; Color; Computer Retrieval of Information on Scientific Projects Database; Core Facility; Cryptosporidiosis; Cryptosporidium; Cryptosporidium parvum; Culicidae; Dengue; Development; Enrollment; Evaluation; Flow Cytometry; Funding; Future; Genetic Predisposition to Disease; Grant; Human; Immune response; Immunity; Immunology; Infection; Institution; Life; Lymphocyte; Lysosomes; Mannose Binding Lectin; Mannose-Binding Lectins; Measures; Memory B-Lymphocyte; Modeling; Paper; Peripheral; Persons; Phagocytosis; Population; Research; Research Personnel; Resources; Salmonella typhi; Serotyping; Serum; Source; Specimen; St. Louis Encephalitis; Staging; Standardization; Uganda; United States National Institutes of Health; Vaccination; Vaccines; Vermont; Vero Cells; Viral; West Nile virus; Work; Yellow fever virus; cohort; cytokine; enzyme linked immunospot assay; genome wide association study; human DNA; indexing; killings; pathogen; research study; surfactant

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. ***Please note the Tables and Figures mentioned below would not reproduce in this format. Please see attachments sent with the paper copy.*** Work in year 4 of the COBRE was focused on exploring new avenues for human cryptosporidiosis research as well as development of new immunology assays to compliment this upcoming work. Establishment of functional immunology assays: Using specimens from clinical research studies on Cryptosporidium and live attenuated S. Typhi vaccines, we have established functional antibody and cellular assays for evaluation of human pathogens and toward standardization for use in a future human immunology core facility. Four new functional antibody assays have been developed using S. typhi as a model pathogen: opsonization/ phagocytosis (Figure 1); bacterial killing post-phagocytosis; measures of phagocytic index (PI, Figure 2); and a lysosome-colocalization assay. For cellular immunology, we have developed a seven-color flow cytometry assay to evaluate intracellular cytokines pre-and post-infection or vaccination (Figure 3). A B-cell memory ELISPOT assay has also been formally developed. All these assays are now in use for studying live attenuated dengue vaccines and will be used for future study of Cryptosporidium infection. Other assays, specifically for the study of viral pathogens, have been developed. These include standardized plaque reduction and neutralization assays (PRNT) using Vero cells (mosquito cell line) for Dengue serotype 1-4, West Nile, St. Louis Encephalitis and Yellow Fever viruses. Viral amplication assays (Dengue serotypes 1-4) have also been developed. New Cryptosporidium studies: opportunities for further Cryptosporidium work are anticipated to come to fruition in year 5 of the COBRE. First, opportunities for Cryptosporidium immunology are available in Dhaka, Bangladesh as part of a large birth cohort of children. Presently, 400 children have been enrolled and 12% have evidence of Cryptosporidium infection by 6 months of life. Clinical data, human DNA, peripheral lymphocytes and sera are available, making this an ideal setting for continuing our work into human immune responses to this infection. The work from this cohort will focus on three major avenues: genome-wide associations to expand our observations of genetic susceptibility to cryptosporidiosis; expanded and functional work on mannose-binding lectin (MBL) and other collectins/surfactants which function as an innate components in immunity to Cryptosporidium infection and evaluation of cellular immune responses to infection using 7 or 8-color flow cytometry and ELISPOT assays (see above).Secondly, we are initiating enrollment of a large cohort of persons with end-stage AIDS and Cryptosporidium infection (Uganda). We will be confirming the MBL association with Cryptosporidium in this population.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 ***请注意，以下提到的表和数字不会以这种格式复制。 请参阅带有纸质副本的附件。*** 毛tr的第4年的工作着重于探索人类隐孢子虫病研究的新途径以及开发新的免疫学分析，以补充这项即将到来的工作。 建立功能免疫学分析：使用有关隐孢子虫和活伤性链球菌疫苗的临床研究的标本，我们已经建立了功能抗体和细胞测定法，以评估人类病原体和标准化，以在未来的人类免疫学核心方面使用。使用S. typhi作为模型病原体开发了四种新的功能抗体测定：调理/吞噬作用（图1）；细菌杀死后刺伤；吞噬指数的度量（PI，图2）；和溶酶体 - 聚自定义测定法。对于细胞免疫学，我们已经开发了一种七色流式细胞仪测定法，以评估细胞内细胞因子的感染前后或疫苗接种（图3）。 B细胞存储器ELISPOT分析也已正式开发。现在，所有这些测定法已用于研究活衰减的登革热疫苗，并将用于未来的隐孢子虫感染研究。已经开发了其他专门用于病毒病原体研究的测定法。其中包括使用Vero细胞（蚊子细胞系）1-4，西尼罗河，圣路易斯脑炎和黄热病毒的标准化斑块还原和中和测定（PRNT）。还开发了病毒扩增测定（登革热血清型1-4）。 新的隐孢子虫研究：预计在鞋底第5年实现进一步的隐孢子虫工作的机会。首先，作为大量儿童的一部分，孟加拉国达卡有隐孢子虫免疫学的机会。目前，已有400名儿童被招募，有12％的儿童在6个月的生命中有隐孢子虫感染的证据。提供临床数据，人DNA，外周淋巴细胞和血清，这是继续我们对这种感染的人类免疫反应的工作的理想场所。该队列的工作将集中在三大途径上：全基因组的关联，以扩大我们对隐孢子虫病的遗传易感性的观察；在甘露糖结合凝集素（MBL）和其他集群蛋白/表面活性剂上的扩展和功能性工作，它们是对隐孢子虫感染免疫的先天成分，并使用7或8色的流式细胞仪和ELISPOT分析（请参阅上述）启动，启动的人群启动了一个较大的人群。隐孢子虫感染（乌干达）。我们将在该人群中确认与隐孢子虫的MBL关联。