Fulltext

Harmonization guidelines for HLA-peptide multimer assays derived from results of a large scale international proficiency panel of the Cancer Vaccine Consortium

Britten, Cedrik ; Janetzki, Sylvia ; Ben-Porat, Leah ; Clay, Timothy ; Kalos, Michael ; Maecker, Holden ; Odunsi, Kunle ; Pride, Michael ; Old, Lloyd ; Hoos, Axel ; Romero, Pedro

In: Cancer Immunology, Immunotherapy, 2009, vol. 58, no. 10, p. 1701-1713

Ajouter à la liste personnelle
    Titre
    • Harmonization guidelines for HLA-peptide multimer assays derived from results of a large scale international proficiency panel of the Cancer Vaccine Consortium
    Auteur
    • Britten, Cedrik. Tumor Immunology Group, Department for Immunohematology and Blood Transfusion, Leiden University Medical Centre, Leiden, The Netherlands
    • Janetzki, Sylvia. ZellNet Consulting, Inc., Fort Lee, NJ, USA
    • Ben-Porat, Leah. Department of Biostatistics, New York University, New York, NY, USA
    • Clay, Timothy. Surgery and Immunology, Duke University Medical Center, Durham, NC, USA
    • Kalos, Michael. Clinical Immunobiology Correlative Studies Laboratory, City of Hope, Duarte, CA, USA
    • Maecker, Holden. BD Biosciences, San Jose, CA, USA
    • Odunsi, Kunle. Departments of Gynecologic Oncology and Immunology, Roswell Park Cancer Institute, Buffalo, NY, USA
    • Pride, Michael. Vaccines Early Phase Programs, Wyeth Research, Pearl River, NY, USA
    • Old, Lloyd. Ludwig Institute for Cancer Research, New York Branch, Memorial Sloan-Kettering Cancer Center, New York, NY, USA
    • Hoos, Axel. Bristol-Myers Squibb, Wallingford, CT, USA
    • Romero, Pedro. Division of Clinical Onco-Immunology, Lausanne Branch, Ludwig Institute for Cancer Research, University Hospital (CHUV), Lausanne, Switzerland
    Type de document
    • Postprint
    Langue
    • Anglais
    Publié dans
    • Cancer Immunology, Immunotherapy, 2009, vol. 58, no. 10, p. 1701-1713. Springer-Verlag
    Autre version électronique
    Identifiant OAI-PMH
    • oai:doc.rero.ch:321713
    Summary
    Purpose: The Cancer Vaccine Consortium of the Cancer Research Institute (CVC-CRI) conducted a multicenter HLA-peptide multimer proficiency panel (MPP) with a group of 27 laboratories to assess the performance of the assay. Experimental design: Participants used commercially available HLA-peptide multimers and a well characterized common source of peripheral blood mononuclear cells (PBMC). The frequency of CD8+ T cells specific for two HLA-A2-restricted model antigens was measured by flow cytometry. The panel design allowed for participants to use their preferred staining reagents and locally established protocols for both cell labeling, data acquisition and analysis. Results: We observed significant differences in both the performance characteristics of the assay and the reported frequencies of specific T cells across laboratories. These results emphasize the need to identify the critical variables important for the observed variability to allow for harmonization of the technique across institutions. Conclusions: Three key recommendations emerged that would likely reduce assay variability and thus move toward harmonizing of this assay. (1) Use of more than two colors for the staining (2) collect at least 100,000 CD8 T cells, and (3) use of a background control sample to appropriately set the analytical gates. We also provide more insight into the limitations of the assay and identified additional protocol steps that potentially impact the quality of data generated and therefore should serve as primary targets for systematic analysis in future panels. Finally, we propose initial guidelines for harmonizing assay performance which include the introduction of standard operating protocols to allow for adequate training of technical staff and auditing of test analysis procedures