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Enzyme-linked immunospot, cytokine flow cytometry, and tetramers in the detection of T-cell responses to a dendritic cell-based multipeptide vaccine in patients with melanoma.
Clin Cancer Res 2003; 9(2):641-9CC

Abstract

A new generation of single-cell assays for measuring the frequency of peptide-specific T lymphocytes in cellular populations has become widely available. These assays, enzyme-linked immunospot (ELISPOT), cytokine flow cytometry, and tetramer binding, are used frequently for monitoring of cancer patient responses to vaccination therapies. We concomitantly used these three assays to determine the frequency of CD8(+) T cells in the circulation of 8 patients with metastatic melanoma who had received a multiepitope peptide/dendritic cell-based vaccine. Using peripheral blood mononuclear cells harvested before and a week after the last of four vaccines, we observed that the three assays detected a substantially different frequency of peptide-specific CD8(+) T cells. The tetramer assay consistently detected the highest numbers of peptide-specific CD8(+) T cells, followed by cytokine flow cytometry and then ELISPOT. There was no significant concordance among the assays in measuring the numbers of CD8(+) T cells specific for each of the peptides in the peripheral circulation of the patient. No significant pre- to postvaccine changes in the number of CD8(+) T cells specific for any of the peptides were observed. Thus, the dendritic cell-based vaccine was observed not to augment immune responses to the peptides in the patients. Because of a low frequency of the peptide-specific T cells in the peripheral circulation of the patients, these sensitive single-cell assays were used to measure values at the lower limit of detection. For this and other reasons, including the issues of tetramer specificity and ELISPOT sensitivity, caution in interpretation and serious attention to quality control are needed in monitoring of immune responses to anticancer vaccines.

Authors+Show Affiliations

University of Pittsburgh Cancer Institute, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15213, USA. whitesidetl@msx.upmc.eduNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Clinical Trial
Clinical Trial, Phase I
Clinical Trial, Phase II
Journal Article
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

12576430

Citation

Whiteside, Theresa L., et al. "Enzyme-linked Immunospot, Cytokine Flow Cytometry, and Tetramers in the Detection of T-cell Responses to a Dendritic Cell-based Multipeptide Vaccine in Patients With Melanoma." Clinical Cancer Research : an Official Journal of the American Association for Cancer Research, vol. 9, no. 2, 2003, pp. 641-9.
Whiteside TL, Zhao Y, Tsukishiro T, et al. Enzyme-linked immunospot, cytokine flow cytometry, and tetramers in the detection of T-cell responses to a dendritic cell-based multipeptide vaccine in patients with melanoma. Clin Cancer Res. 2003;9(2):641-9.
Whiteside, T. L., Zhao, Y., Tsukishiro, T., Elder, E. M., Gooding, W., & Baar, J. (2003). Enzyme-linked immunospot, cytokine flow cytometry, and tetramers in the detection of T-cell responses to a dendritic cell-based multipeptide vaccine in patients with melanoma. Clinical Cancer Research : an Official Journal of the American Association for Cancer Research, 9(2), pp. 641-9.
Whiteside TL, et al. Enzyme-linked Immunospot, Cytokine Flow Cytometry, and Tetramers in the Detection of T-cell Responses to a Dendritic Cell-based Multipeptide Vaccine in Patients With Melanoma. Clin Cancer Res. 2003;9(2):641-9. PubMed PMID: 12576430.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Enzyme-linked immunospot, cytokine flow cytometry, and tetramers in the detection of T-cell responses to a dendritic cell-based multipeptide vaccine in patients with melanoma. AU - Whiteside,Theresa L, AU - Zhao,Yongxiang, AU - Tsukishiro,Takashi, AU - Elder,Elaine M, AU - Gooding,William, AU - Baar,Joseph, PY - 2003/2/11/pubmed PY - 2003/8/2/medline PY - 2003/2/11/entrez SP - 641 EP - 9 JF - Clinical cancer research : an official journal of the American Association for Cancer Research JO - Clin. Cancer Res. VL - 9 IS - 2 N2 - A new generation of single-cell assays for measuring the frequency of peptide-specific T lymphocytes in cellular populations has become widely available. These assays, enzyme-linked immunospot (ELISPOT), cytokine flow cytometry, and tetramer binding, are used frequently for monitoring of cancer patient responses to vaccination therapies. We concomitantly used these three assays to determine the frequency of CD8(+) T cells in the circulation of 8 patients with metastatic melanoma who had received a multiepitope peptide/dendritic cell-based vaccine. Using peripheral blood mononuclear cells harvested before and a week after the last of four vaccines, we observed that the three assays detected a substantially different frequency of peptide-specific CD8(+) T cells. The tetramer assay consistently detected the highest numbers of peptide-specific CD8(+) T cells, followed by cytokine flow cytometry and then ELISPOT. There was no significant concordance among the assays in measuring the numbers of CD8(+) T cells specific for each of the peptides in the peripheral circulation of the patient. No significant pre- to postvaccine changes in the number of CD8(+) T cells specific for any of the peptides were observed. Thus, the dendritic cell-based vaccine was observed not to augment immune responses to the peptides in the patients. Because of a low frequency of the peptide-specific T cells in the peripheral circulation of the patients, these sensitive single-cell assays were used to measure values at the lower limit of detection. For this and other reasons, including the issues of tetramer specificity and ELISPOT sensitivity, caution in interpretation and serious attention to quality control are needed in monitoring of immune responses to anticancer vaccines. SN - 1078-0432 UR - https://www.unboundmedicine.com/medline/citation/12576430/Enzyme_linked_immunospot_cytokine_flow_cytometry_and_tetramers_in_the_detection_of_T_cell_responses_to_a_dendritic_cell_based_multipeptide_vaccine_in_patients_with_melanoma_ L2 - http://clincancerres.aacrjournals.org/cgi/pmidlookup?view=long&pmid=12576430 DB - PRIME DP - Unbound Medicine ER -