CHARACTERIZATION OF A PROTECTIVE ANTIGEN OF T GONDII

弓形虫保护性抗原的表征

基本信息

批准号：
3133868
负责人：
FAUSTO G ARAUJO
金额：
$ 14.41万
依托单位：
PALO ALTO MEDICAL FOUNDATION RES INST
依托单位国家：
美国
项目类别：
财政年份：
1986
资助国家：
美国
起止时间：
1986-03-01 至 1989-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/3133868
关键词：
Toxoplasma gondii enzyme linked immunosorbent assay gel electrophoresis genetic manipulation genetic regulation host organism interaction immunochemistry interferons laboratory mouse microorganism immunology monoclonal antibody protein structure protozoal antigen toxoplasmosis

项目摘要

The long term goal of the proposed research is to isolate and characterize a cytoplasmic antigen that stimulates protective immunity to an intracellular parasite Toxoplasma gondii. We will further define the specificity of the antigen purified from an affinity chromatography column with the aid of western blotting and enzyme linked immunosorbent assays. The molecular weight of the antigen will be determined using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Experiments will be performed to determine whether the purified antigen is a protein or a glycoprotein. Chemical and proteolytic cleavage of the purified antigen will be carried out to define the location of the epitopes recognized by the monoclonal antibody. We will clone the gene coding for this protein and develop methodology to obtain large quantities of this antigen. Studies will be conducted to define the mechanisms by which purified antigen, antigenic fragments obtained as a result of proteolytic and chemical cleavage as well as the antigen produced by recombinant DNA methods induces a protective immune response. The ability of these antigenic preparations to prime spleen cells to secrete interferon, activate macrophages, produce different classes of immunoglobulins and affect the number of cysts formed will be examined. We hope the proposed research will lead to a better understanding of the nature of the antigenic structure of Toxoplasma and the mechanisms by which well characterized antigens stimulate an immune response beneficial to the host. Our gene cloning experiments will not only allow us to produce large quantities of the antigen but will also open the doors for future studies concerned with the regulation of genes coding for the antigens of Toxoplasma gondii.

拟议研究的长期目标是分离和表征刺激保护性免疫的细胞质抗原细胞内寄生虫弓形虫。我们将进一步明确从亲和层析柱纯化的抗原的特异性借助蛋白质印迹和酶联免疫吸附测定。使用十二烷基钠测定抗原的分子量硫酸盐-聚丙烯酰胺凝胶电泳。将进行实验以确定纯化的抗原是蛋白质还是糖蛋白。将进行纯化抗原的化学和蛋白水解切割确定单克隆抗体识别的表位的位置抗体。我们将克隆编码该蛋白质的基因并开发获得大量该抗原的方法。研究将是进行以确定纯化的抗原、抗原性的机制由于蛋白水解和化学切割而获得的片段因为重组 DNA 方法产生的抗原会诱导保护性免疫反应。这些抗原制剂的引发能力脾细胞分泌干扰素，激活巨噬细胞，产生不同的免疫球蛋白的类别并影响形成的包囊数量检查了。我们希望所提出的研究能够带来更好的结果了解弓形虫抗原结构的性质和明确表征的抗原刺激免疫的机制对宿主有利的反应。我们的基因克隆实验不会只允许我们生产大量抗原，但也会开放未来有关基因编码调控的研究之门为弓形虫抗原。