In situ visualization of antigen-specific T cells in cryopreserved human tissues

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• 作者：Florry A. Vyth-Dreese ; Yeung-Hyen Kim ; Trees A.M. Dellemijn ; Ellen Schrama ; John B.A.G. Haanen ; Eric Spierings and Els Goulmy
• 关键词：Humans ; CD8-positive T-lymphocytes/immunology ; Major Histocompatibility antigens class I ; Minor Histocompatibility antigens ; In situ ; Ex vivo ; Staining and labeling methods ; Fluorescent Dyes
• 刊名：Journal of Immunological Methods
• 出版年：2006
• 出版时间：20 March 2006
• 年：2006
• 卷：310
• 期：1-2
• 页码：78-85
• 全文大小：354 K

文摘

Tetrameric MHC/peptide complexes are important tools for analyzing antigen-specific T cells. The in situ use of tetrameric MHC/peptide complexes in viable tissue sections has several shortcomings: it does not allow the execution of multiple analyses on one single biopsy, the storage of the biopsies, and the co-staining of the tetramer-positive cells for various intracellular molecules. We have developed a novel approach using overnight pre-labeling of viable human tissues with MHC/peptide tetramers, followed by cryopreservation and labeling of the cryosections. The visualization of antigen-specific T cells, combined with detection of other membrane, cytoplasmic, or nuclear markers is now feasible.