Validation of a HLA-A2 tetramer flow cytometric method, IFNgamma real time RT-PCR, and IFNgamma ELISPOT for detection of immunologic response to gp100 and MelanA/MART-1 in melanoma patients
Open Access
- 22 October 2008
- journal article
- research article
- Published by Springer Nature in Journal of Translational Medicine
- Vol. 6 (1) , 61
- https://doi.org/10.1186/1479-5876-6-61
Abstract
HLA-A2 tetramer flow cytometry, IFNγ real time RT-PCR and IFNγ ELISPOT assays are commonly used as surrogate immunological endpoints for cancer immunotherapy. While these are often used as research assays to assess patient's immunologic response, assay validation is necessary to ensure reliable and reproducible results and enable more accurate data interpretation. Here we describe a rigorous validation approach for each of these assays prior to their use for clinical sample analysis.Keywords
This publication has 23 references indexed in Scilit:
- Targeting the local tumor microenvironment with vaccinia virus expressing B7.1 for the treatment of melanomaJournal of Clinical Investigation, 2005
- Standardization and Validation Issues of the ELISPOT AssayPublished by Springer Nature ,2005
- Autoimmunity in a Phase I Trial of a Fully Human Anti-Cytotoxic T-Lymphocyte Antigen-4 Monoclonal Antibody With Multiple Melanoma Peptides and Montanide ISA 51 for Patients With Resected Stages III and IV MelanomaJournal of Clinical Oncology, 2005
- Evaluation of Elispot assays: influence of method and operator on variability of resultsJournal of Immunological Methods, 2004
- Analysis of T-cell responses in metastatic melanoma patients vaccinated with dendritic cells pulsed with tumor lysatesCancer Immunology, Immunotherapy, 2004
- Ex Vivo Analysis of Human Antigen-Specific CD8+ T-Cell Responses: Quality Assessment of Fluorescent HLA-A2 Multimer and Interferon-γ ELISPOT Assays for Patient Immune MonitoringJournal of Immunotherapy, 2004
- A phase I trial of SD-9427 (progenipoietin) with a multipeptide vaccine for resected metastatic melanoma.2003
- Immunologic Monitoring of Cancer Vaccine Therapy: Results of a Workshop Sponsored by the Society for Biological TherapyJournal of Immunotherapy, 2002
- Peptide‐specific CD8+ T‐cell evolution in vivo: Response to peptide vaccination with Melan‐A/MART‐1International Journal of Cancer, 2002
- Functional Characterization of CD8+ Antigen-Specific Cytotoxic T Lymphocytes after Enrichment Based on Cytokine Secretion: Comparison with the MHC-Tetramer TechnologyScandinavian Journal of Immunology, 2000