Ex vivo identification, isolation and analysis of tumor-cytolytic T cells

Valerie Rubio, Tor B. Stuge, Naileshni S Singh, Michael R. Betts, Jeffrey S. Weber, Mario Roederer, Peter P. Lee

Research output: Contribution to journalArticle

316 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)1377-1382
Number of pages6
JournalNature Medicine
Volume9
Issue number11
DOIs
StatePublished - Nov 2003
Externally publishedYes

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T-cells
Tumors
T-Lymphocytes
Neoplasms
Peptides
Bearings (structural)
Cancer Vaccines
Vaccination
Peptide T
Adoptive Immunotherapy
Immunologic Monitoring
Membrane Proteins
Blood
Major Histocompatibility Complex
Immunotherapy
Antigens
Monitoring
Staining and Labeling

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Rubio, V., Stuge, T. B., Singh, N. S., Betts, M. R., Weber, J. S., Roederer, M., & Lee, P. P. (2003). Ex vivo identification, isolation and analysis of tumor-cytolytic T cells. Nature Medicine, 9(11), 1377-1382. https://doi.org/10.1038/nm942

Ex vivo identification, isolation and analysis of tumor-cytolytic T cells. / Rubio, Valerie; Stuge, Tor B.; Singh, Naileshni S; Betts, Michael R.; Weber, Jeffrey S.; Roederer, Mario; Lee, Peter P.

In: Nature Medicine, Vol. 9, No. 11, 11.2003, p. 1377-1382.

Research output: Contribution to journalArticle

Rubio, V, Stuge, TB, Singh, NS, Betts, MR, Weber, JS, Roederer, M & Lee, PP 2003, 'Ex vivo identification, isolation and analysis of tumor-cytolytic T cells', Nature Medicine, vol. 9, no. 11, pp. 1377-1382. https://doi.org/10.1038/nm942
Rubio V, Stuge TB, Singh NS, Betts MR, Weber JS, Roederer M et al. Ex vivo identification, isolation and analysis of tumor-cytolytic T cells. Nature Medicine. 2003 Nov;9(11):1377-1382. https://doi.org/10.1038/nm942
Rubio, Valerie ; Stuge, Tor B. ; Singh, Naileshni S ; Betts, Michael R. ; Weber, Jeffrey S. ; Roederer, Mario ; Lee, Peter P. / Ex vivo identification, isolation and analysis of tumor-cytolytic T cells. In: Nature Medicine. 2003 ; Vol. 9, No. 11. pp. 1377-1382.
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