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

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State: Public
Version: author
Serval ID
serval:BIB_03073F7A73B1
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Harmonization guidelines for HLA-peptide multimer assays derived from results of a large scale international proficiency panel of the Cancer Vaccine Consortium.
Journal
Cancer Immunology, Immunotherapy
Author(s)
Britten C.M., Janetzki S., Ben-Porat L., Clay T.M., Kalos M., Maecker H., Odunsi K., Pride M., Old L., Hoos A., Romero P.
Working group(s)
HLA-peptide Multimer Proficiency Panel of the CVC-CRI Immune Assay Working Group
Contributor(s)
Ahmed RK., Bain C., Barnby-Porritt H., Baumgartner P., Bercovici N., Brockstedt D., Cerundolo V., Channon JY., Chikoti P., Cox J., Currier JR., Curtsinger J., Desmaretz C., Gagnon D., Gearhart J., Gillanders V., Hampl J., Harrop R., Hobeika A., Kaempgen E., Kaufman J., Labroquere K., Landry C., Maeurer M., Matijevic M., Mueller S., Mohanakumar T., Olson W., Pohla H., Ramesh R., Rooke R., Samorski R., Sékaly RP., Schullery D., Slingluff C., Scheibenbogen C., Schmitt-Händle M., Smith K., Speiser D., Tarlton A., Trautmann L., Van Der Aa A., Walter S., Wolchok J., Yuan J.
ISSN
1432-0851 (Electronic)
ISSN-L
0340-7004
Publication state
Published
Issued date
2009
Peer-reviewed
Oui
Volume
58
Number
10
Pages
1701-1713
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: ppublish
Abstract
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.
Keywords
Biological Assay, Cancer Vaccines/immunology, Clinical Laboratory Techniques/standards, Guidelines as Topic, HLA-A2 Antigen/immunology, HLA-A2 Antigen/metabolism, Humans, International Cooperation, Peptide Fragments/immunology, Peptide Fragments/metabolism, Protein Multimerization
Pubmed
Web of science
Open Access
Yes
Create date
15/01/2010 14:56
Last modification date
20/08/2019 12:25
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