ASSAY FOR HAPTEN SPECIFIC PRIMING OF T LYMPHOCYTES

T 淋巴细胞半抗原特异性启动的测定

• 批准号: 6141416
• 负责人: J WAYNE STREILEIN
• 金额: $ 4.05万
• 依托单位: SCHEPENS EYE RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-09-30 至 2000-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6141416
• 关键词: T lymphocyte; bioassay; cell proliferation; clinical research; contact dermatitis; cytokine; cytotoxicity; haptens; human subject; pathologic process; skin hypersensitivity; skin irritation /irritant; tissue /cell culture

DESCRIPTION: (Adapted from the Investigator's Abstract) Allergic and irritant contact dermatitis (ACD and ICD) are common inflammatory skin conditions that result after exposure of the cutaneous surface to certain environmental agents (contact sensitizers or haptens). Lack of clear understanding of immunopathogenesis of ACD so far has remained the major hindrance in achieving cure or prevention of these disorders. Effort towards prevention of ACD include identification of environmental agents that induce contact hypersensitivity (CH) response as well as individuals that are susceptible for such a response. Although it is known that certain universal contact sensitizing agents are chemically reactive low molecular weight compounds that can derivatize proteins upon contact and become immunogenic, it is not known why some contact sensitizers do not induce CH in all the individuals. Also, currently available means for the identification of contact allergens can not discriminate between allergens and irritants. In vitro assays designed to identify contact sensitizers so far have met with limited success. These assays have relied on the proliferative responses of T cells, which is a predominant characteristic of a CH response besides inflammation. However, T cell proliferation has remained an inconsistent indicator for distinguishing haptens from irritants. Based on the distinct sensitizing properties of haptens as compared to irritants we propose to develop an in vitro assay that may help identify potential contact sensitizers and irritants as well as individuals susceptible to developing ACD or ICD. We have so far successfully demonstrated the possibility of generating hapten-specific sensitized T cells in vitro cultures in which autologous naive T cells are exposed to hapten derivatized cultured PBMC for 5 days. In order to generate sensitized T cells that closely resemble those found in vivo we now plan to compare effector functions of T cells primed in vivo and in vitro. These comparisons will include phenotype of the T cells proliferating in a hapten-specific manner, their cytokine profile and hapten-specific cytotoxicity. Attempts will be made to compare and contrast T cell responses to various haptens and irritants. Thus, this project can help (a) refine an in vitro assay for future application in clinical laboratory for screening various potential haptens and irritants as well as susceptible individuals and (b) provide better understanding of cellular events that lead to allergic versus irritant contact dermatitis.

描述：（改编自研究者摘要）过敏和 刺激性接触性皮炎（ACD和ICD）是常见炎症性皮肤 在皮肤表面暴露于某些环境中后产生的病症 环境因素（接触致敏剂或半抗原）。 缺乏明确的 到目前为止，对ACD免疫发病机制的理解仍然是主要的 阻碍实现这些病症治愈或预防。 努力 包括识别环境因子 引起接触性超敏反应（CH）以及个体 容易受到这种反应的影响。 尽管众所周知， 通用接触敏化剂是化学反应性低分子的 这些化合物可以在接触时衍生蛋白质， 由于具有免疫原性，尚不清楚为什么某些接触性致敏剂不诱导CH 在所有的个体中。 此外，目前可用于 接触性过敏原鉴定不能区分过敏原 和刺激物。 体外试验旨在识别接触致敏物， 迄今为止，取得了有限的成功。 这些分析依赖于 T细胞的增殖反应，这是一个主要特征， 除了炎症还有CH反应 然而，T细胞增殖具有 仍然是区分半抗原和 刺激物。 基于半抗原的不同致敏特性， 与刺激物相比，我们建议开发一种体外测定方法， 识别潜在的接触致敏物和刺激物以及个体 易患ACD或ICD。 到目前为止，我们已经成功地 证明了产生半抗原特异性致敏T细胞的可能性。 细胞在体外培养物中，其中自体幼稚T细胞暴露于 半抗原衍生化培养的PBMC 5天。 以便生成 致敏的T细胞与体内发现的非常相似，我们现在计划 比较体内和体外致敏的T细胞的效应子功能。 这些 比较将包括T细胞的表型增殖， 半抗原特异性方式，它们的细胞因子谱和半抗原特异性 细胞毒 将尝试比较和对比T细胞 对各种半抗原和刺激物的反应。 因此，该项目可以帮助（a） 改进体外测定法，以便将来在临床实验室中应用， 筛选各种潜在的半抗原和刺激物以及敏感的 和（B）提供对细胞事件的更好理解， 导致过敏性对刺激性接触性皮炎。