Nature Medicine9, 1377 - 1382 (2003)
Published online: 5 October 2003; | doi:10.1038/nm942
Ex vivo identification, isolation and analysis of tumor-cytolytic T cells
Valerie Rubio1, Tor B Stuge1, Naileshni Singh1, Michael R Betts2, Jeffrey S Weber3, Mario Roederer4
& Peter P Lee1
1
Department of Medicine, Stanford University, 269 Campus Drive, Stanford, California 94305, USA.
2
Laboratory of Immunology, Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 40 Convent Drive, Bethesda, Maryland 20892, USA.
3
Norris Cancer Center, University of Southern California, 1441 Eastlake Avenue, Suite 3454, Los Angeles, California 90033, USA.
4
ImmunoTechnology Section, Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 40 Convent Drive, Bethesda, Maryland 20892, USA.
Correspondence should be addressed to Peter P Lee ppl@stanford.edu
We isolated pure, viable populations of tumor-cytolytic T cells directly from patient blood samples using flow cytometric quantification of the surface mobilization of CD107a—an integral membrane protein in cytolytic granules—as a marker for degranulation after tumor stimulation. We show that tumor-cytolytic T cells are indeed elicited in patients after cancer vaccination, and that tumor reactivity is strongly correlated with efficient T-cell recognition of peptide-bearing targets. We combined CD107a mobilization with peptide−major histocompatibility complex (P-MHC) tetramer staining to directly correlate antigen specificity and cytolytic ability on a single-cell level. This showed that tumor-cytolytic T cells with high recognition efficiency represent only a minority of peptide-specific T cells elicited in patients after heteroclitic peptide vaccination. We were also able to expand these cells to high numbers ex vivo while maintaining their cytolytic potential. These techniques will be useful not only for immune monitoring of cancer vaccine trials, but also for adoptive cellular immunotherapy after ex vivo expansion. The ability to rapidly identify and isolate tumor-cytolytic T cells would be very useful in cancer immunotherapy.
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