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
- PMID: 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
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.
Similar articles
-
Immune modulations during chemoimmunotherapy & novel vaccine strategies--in metastatic melanoma and non small-cell lung cancer.Dan Med J. 2013 Dec;60(12):B4774. Dan Med J. 2013. PMID: 24355457 Review.
-
Analysis and characterization of antitumor T-cell response after administration of dendritic cells loaded with allogeneic tumor lysate to metastatic melanoma patients.J Immunother. 2008 Jan;31(1):101-12. doi: 10.1097/CJI.0b013e318159f5ba. J Immunother. 2008. PMID: 18157017 Clinical Trial.
-
Immune responses to a class II helper peptide epitope in patients with stage III/IV resected melanoma.Clin Cancer Res. 2004 Aug 1;10(15):5004-13. doi: 10.1158/1078-0432.CCR-04-0241. Clin Cancer Res. 2004. PMID: 15297401 Clinical Trial.
-
How to analyze ex vivo T-cell responses in cancer patients.In Vivo. 2002 Nov-Dec;16(6):519-25. In Vivo. 2002. PMID: 12494896 Review.
-
Immunization of patients with melanoma peptide vaccines: immunologic assessment using the ELISPOT assay.Cancer J Sci Am. 1998 Sep-Oct;4(5):316-23. Cancer J Sci Am. 1998. PMID: 9815296 Clinical Trial.
Cited by
-
Antibody Engineering for Pursuing a Healthier Future.Front Microbiol. 2017 Mar 28;8:495. doi: 10.3389/fmicb.2017.00495. eCollection 2017. Front Microbiol. 2017. PMID: 28400756 Free PMC article. Review.
-
Cellular immunity induced by a recombinant adenovirus- human dendritic cell vaccine for melanoma.J Immunother Cancer. 2013 Nov 18;1:19. doi: 10.1186/2051-1426-1-19. eCollection 2013. J Immunother Cancer. 2013. PMID: 24829755 Free PMC article.
-
Testing for HLA/peptide tetramer-binding to the T cell receptor complex on human T lymphocytes.Results Immunol. 2012 May 10;2:88-96. doi: 10.1016/j.rinim.2012.04.001. eCollection 2012. Results Immunol. 2012. PMID: 24371571 Free PMC article.
-
Anti-leukemia T cells in AML: TNF-α⁺ CD8⁺ T cells may escape detection and possibly reflect a stage of functional impairment.Hum Vaccin Immunother. 2013 Jun;9(6):1200-4. doi: 10.4161/hv.24250. Epub 2013 Apr 8. Hum Vaccin Immunother. 2013. PMID: 23571180 Free PMC article.
-
Immunogenic comparison of chimeric adenovirus 5/35 vector carrying optimized human immunodeficiency virus clade C genes and various promoters.PLoS One. 2012;7(1):e30302. doi: 10.1371/journal.pone.0030302. Epub 2012 Jan 19. PLoS One. 2012. PMID: 22276174 Free PMC article.
Publication types
MeSH terms
Substances
Grants and funding
LinkOut - more resources
Other Literature Sources
Medical
Research Materials