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目的 :研究金黄色葡萄球菌D型肠毒素 (SED)超抗原与TCRVβ结合的关键位点。 方法 :利用定点突变技术 ,构建了 6种SED的突变体。用3 H TdR掺入法检测这些突变体促进T细胞增殖的活性。对促增殖活性降低的突变体 ,进一步用流式细胞仪检测他们与MHC Ⅱ类分子结合的活性和其TCRVβ特异性。结果 :发现N2 3位氨基酸是SED与人TCRVβ5结合的关键位点。H2 6位氨基酸可能是SED与人的其他TCRVβ(除TCRVβ5、TCRVβ8和TCRVβ12 .1)结合的关键位点 ,值得进一步研究。结论 :本结果丰富了对超抗原免疫识别的认识 ,为葡萄球菌超抗原相关疾病的防治 ,以及基于超抗原设计新的抗肿瘤免疫制剂提供了理论依据
Objective: To study the key sites of binding of Staphylococcus aureus D-enterotoxin (SED) superantigen to TCRVβ. Methods: Six SED mutants were constructed by site-directed mutagenesis. These mutants were tested for their ability to promote T cell proliferation by 3 H TdR incorporation. For mutants with reduced proliferative activity, their binding to MHC class II molecules and their TCRVβ specificity were further examined by flow cytometry. Results: The N2 3 amino acid was found to be a key site for the binding of SED to human TCRVβ5. The amino acid at position 6 of H2 may be a key site for SED binding to other human TCRVβ (except TCRVβ5, TCRVβ8 and TCRVβ12.1) and warrants further study. CONCLUSION: This result enriches the understanding of superantigen immune recognition, provides a theoretical basis for the prevention and treatment of staphylococcal superantigen-related diseases and the design of new anti-tumor immune agents based on superantigens