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使用硝酸纤维素免疫印迹法分析T淋巴细胞对抗原的识别。

The use of nitrocellulose immunoblots for the analysis of antigen recognition by T lymphocytes.

作者信息

Lamb J R, O'Hehir R E, Young D B

机构信息

MRC Tuberculosis and Related Infections Unit, Royal Postgraduate Medical School, Hammersmith Hospital, London, U.K.

出版信息

J Immunol Methods. 1988 May 25;110(1):1-10. doi: 10.1016/0022-1759(88)90076-2.

Abstract

Antigen insolubilised on nitrocellulose is able to activate T lymphocytes following presentation by appropriate accessory cells. Furthermore, complex mixtures of antigens can be fractionated by polyacrylamide gel electrophoresis under denaturing conditions (SDS-PAGE), transferred to nitrocellulose membranes and then added to T cell proliferation assays. This allows a direct screening for recognition of individual polypeptides by monoclonal and polyclonal T cell populations, without a requirement for extensive biochemical purification and in the absence of initial serological preselection of antigen. The potential use of this technique for screening of recombinant DNA libraries for expression of antigens recognised by T cells is also illustrated. The technical aspects of T cell recognition of solid-phase antigen and its application to the identification of immunodominant epitopes with the potential to modulate T cell-mediated immunity is reviewed.

摘要

固定在硝酸纤维素上的抗原在由合适的辅助细胞呈递后能够激活T淋巴细胞。此外,抗原的复杂混合物可在变性条件下通过聚丙烯酰胺凝胶电泳(SDS-PAGE)进行分离,转移至硝酸纤维素膜上,然后添加到T细胞增殖试验中。这使得能够直接筛选单克隆和多克隆T细胞群体对单个多肽的识别,而无需进行广泛的生化纯化,且无需对抗原进行初始血清学预选。本文还阐述了该技术在筛选重组DNA文库以寻找T细胞识别的抗原表达方面的潜在用途。本文综述了T细胞对固相抗原的识别技术及其在鉴定具有调节T细胞介导免疫潜力的免疫显性表位中的应用。

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