Expansion of melanoma-reactive T cells from peptide- stimulated PBL of vaccinated patients is oligoclonal in short term cultures

M. D. McKee, T. M. Clay, M. C. Custer, J. Wunderlich, S. A. Rosenberg, M. I. Nishimura

Research output: Contribution to journalArticle

Abstract

The identification and cloning of tumor associated antigens has led to experimental vaccine and adoptive immunotherapy trials in patients with metastatic melanoma. In one adoptive immunotherapy trial, PBMC were collected on 5 consecutive days from a patient vaccinated with the modified melanoma associated antigenic peptide gp100:209-217 2M in IFA. These PBMC were stimulated once in vitro with gp100:209-217 2M peptide and expanded for 14 days with IL-2 for patient treatment. Each bulk culture showed specific reactivity against the native gp100:209-217 and modified gp100:209-21T 2M peptides as well as HLA-A2+ melanoma lines. We evaluated the TCR repertoire in these short term cultures to identify T cell clonotypes which rapidly expanded following peptide stimulation in vitro. Reproducible expansion of 6 of 24 TCRBV sub-families was seen in these cultures using a competitive RT-PCR assay. 5 of the 6 TCRBV subfamilies contained a predominant T cell clonotype, which represented between 2% and 35% of T cells in one short term culture. Limiting dilution cloning from the PBMC of this patient confirmed that at least 2 of the 5 predominant clones recognize the immunizing gp100:209-217 2M peptide. These results suggested that a small set of highly-reactive T cells can be identified from melanoma patients using short term in vitro peptide stimulated cultures. These T cells may be useful for understanding the activation and function of tumor-reactive T cells, monitoring immune therapies, and designing treatment protocols with increased efficacy.

Original languageEnglish (US)
JournalFASEB Journal
Volume12
Issue number5
StatePublished - Mar 20 1998
Externally publishedYes

Fingerprint

T-cells
melanoma
Melanoma
T-lymphocytes
peptides
T-Lymphocytes
Peptides
mononuclear leukocytes
Cell culture
Adoptive Immunotherapy
Cloning
immunotherapy
Organism Cloning
Tumors
molecular cloning
Patient treatment
HLA-A2 Antigen
Immunologic Monitoring
neoplasms
Neoplasm Antigens

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Cell Biology

Cite this

McKee, M. D., Clay, T. M., Custer, M. C., Wunderlich, J., Rosenberg, S. A., & Nishimura, M. I. (1998). Expansion of melanoma-reactive T cells from peptide- stimulated PBL of vaccinated patients is oligoclonal in short term cultures. FASEB Journal, 12(5).

Expansion of melanoma-reactive T cells from peptide- stimulated PBL of vaccinated patients is oligoclonal in short term cultures. / McKee, M. D.; Clay, T. M.; Custer, M. C.; Wunderlich, J.; Rosenberg, S. A.; Nishimura, M. I.

In: FASEB Journal, Vol. 12, No. 5, 20.03.1998.

Research output: Contribution to journalArticle

McKee, MD, Clay, TM, Custer, MC, Wunderlich, J, Rosenberg, SA & Nishimura, MI 1998, 'Expansion of melanoma-reactive T cells from peptide- stimulated PBL of vaccinated patients is oligoclonal in short term cultures', FASEB Journal, vol. 12, no. 5.
McKee MD, Clay TM, Custer MC, Wunderlich J, Rosenberg SA, Nishimura MI. Expansion of melanoma-reactive T cells from peptide- stimulated PBL of vaccinated patients is oligoclonal in short term cultures. FASEB Journal. 1998 Mar 20;12(5).
McKee, M. D. ; Clay, T. M. ; Custer, M. C. ; Wunderlich, J. ; Rosenberg, S. A. ; Nishimura, M. I. / Expansion of melanoma-reactive T cells from peptide- stimulated PBL of vaccinated patients is oligoclonal in short term cultures. In: FASEB Journal. 1998 ; Vol. 12, No. 5.
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