CHARACTERIZATION OF A PROTECTIVE ANTIGEN OF T GONDII

拟议研究的长期目标是分离和表征 一种细胞质抗原，能刺激对免疫系统的保护性免疫， 细胞内寄生虫弓形虫。 我们将进一步定义 亲和层析柱纯化抗原的特异性 借助于蛋白质印迹和酶联免疫吸附测定。 将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， 以确定单克隆抗体识别的表位的位置， 抗体的 我们将克隆编码这种蛋白质的基因， 获得大量这种抗原的方法。 研究将 进行以确定纯化的抗原、抗原性 作为蛋白水解和化学切割的结果获得的片段 因为通过重组DNA方法产生的抗原诱导保护性的 免疫反应 这些抗原制剂的能力， 脾细胞分泌干扰素，激活巨噬细胞，产生不同的 类免疫球蛋白和影响囊肿形成的数量将是 考察 我们希望拟议中的研究将导致更好的 了解弓形虫抗原结构的性质， 充分表征的抗原刺激免疫的机制 对宿主有益的反应。 我们的基因克隆实验 只允许我们生产大量的抗原， 未来研究基因编码调控的大门 弓形虫的抗原

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.