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CHARACTERIZATION OF ANTIGEN-SPECIFIC T CELL PRODUCTS

抗原特异性 T 细胞产品的表征

基本信息

批准号：
3127785
负责人：
ARTHUR MALLEY
金额：
$ 10.24万
依托单位：
OREGON REGIONAL PRIMATE RESEARCH CENTER
依托单位国家：
美国
项目类别：
财政年份：
1983
资助国家：
美国
起止时间：
1983-04-01 至 1986-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/3127785
关键词：
B lymphocyte Gramineae T lymphocyte enzyme linked immunosorbent assay helper T lymphocyte hybridomas hypersensitivity desensitization immunization immunoglobulin G pollen suppressor T lymphocyte tissue /cell culture

项目摘要

We propose to isolate and characterize antigen-B-specific T helper (THF) and T suppressor (TSF) factors from either murine spleen cell populations or appropriate T cell lines or hybridomas. The amount of THF and TSF produced will be quantitated by inhibition of monoclonal anti-AgB antibody binding to AgB bound to microtiter plates (ELISA). Methods used for factor purification will include affinity chromatography, gel-filtration, ion-exchange chromatography, isoelectrofocusing, and high pressure liquid chromatography (HPLC). The purified THF and TSF factors will be used to determine the relationship with immunoglobulin molecules that share antigen-binding properties but otherwise appear structurally unrelated, the nature of the interaction between the antigen-binding and I-region chains, and the role of the carbohydrate moiety in our factors. Another important goal is to develop an immunotherapy model with which to determine the potential of AgB-specific TSF for the treatment of timothy-sensitive patients. These studies will allow us to determine the optimal dose and frequency of TSF treatment to achieve maximal suppression of timothy IgE formation, and to determine the effect of multiple doses of TSF upon the IgE response of hyperimmunized animals. Anti-idiotypic antibodies (anti-THF and anti-IgEid) passed over Sepharose-fetal calf serum, -normal mouse serum, and -F(ab)2 adsorbents and finally eluted from Sepharose-TSF adsorbents will be used to determine the relationship between the idiotype expressed on T and B cells involved in IgG and IgE formation to the same antigenic determinant. Experiments to develop a tissue culture method to study the in vitro synthesis of murine IgE will permit us to examine the mechanism of IgE regulation by isolated T cell factors (THF, TSF1, and TSF2), and examine the site of action of these factors. The IgE recovered from future supernatants will be assayed by an ELISA method using a fluorescent substrate.

我们建议分离和鉴定抗原B特异性辅助性T细胞（THF）和T抑制因子（TSF）或合适的T细胞系或杂交瘤。 THF和TSF的量将通过单克隆抗AgB抗体的抑制来定量产生的抗体与结合于微量滴定板的AgB的结合（ELISA）。用于因子的方法纯化包括亲和层析，凝胶过滤，离子交换层析、等电聚焦和高压液相色谱法（HPLC）。纯化的THF和TSF因子将用于确定与免疫球蛋白分子的关系，抗原结合性质，但在其他方面似乎结构无关，抗原结合链和I区链之间相互作用的性质，以及碳水化合物在我们因子中的作用。另一个重要目标是开发一种免疫治疗模型， AgB特异性TSF治疗胸腺敏感性患者这些研究将使我们能够确定最佳剂量， TSF治疗频率，以达到最大程度抑制Timothy IgE 形成，并确定多剂量的TSF对超免疫动物的IgE应答。抗独特型抗体（抗-THF和抗-IgEid）通过琼脂糖-胎牛血清，-正常小鼠血清和-F（ab）2吸附剂，最后从Sepharose-TSF洗脱吸附剂将被用来确定独特型之间的关系，在参与IgG和IgE形成的T和B细胞上表达抗原决定簇开发组织培养方法的实验研究鼠IgE的体外合成将使我们能够检查分离的T细胞因子（THF、TSF 1和 TSF 2），并检查这些因素的作用部位。 IgE恢复将通过ELISA方法，使用荧光底物。