In situ visualization of antigen-specific T cells in cryopreserved human tissues.
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.
Division of Immunology, The Netherlands Cancer Institute, Amsterdam.
Florry A
FA
Vyth-Dreese
Yeung-Hyen
YH
Kim
Trees A M
TA
Dellemijn
Ellen
E
Schrama
John B A G
JB
Haanen
Eric
E
Spierings
Els
E
Goulmy
D001706:Biopsy; D015925:Cryopreservation; D005260:Female; D005456:Fluorescent Dyes / Q000737:chemistry; D006182:H-Y Antigen / Q000032:analysis / Q000276:immunology; D015789:HLA-A2 Antigen / Q000032:analysis / Q000276:immunology; D006801:Humans; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D008297:Male; D012867:Skin / Q000166:cytology / Q000276:immunology*; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology*
D005456:Fluorescent Dyes; D006182:H-Y Antigen; D015789:HLA-A2 Antigen