Evaluation of the Interlaboratory Concordance in Quantification of Human Immunodeficiency Virus-Specific T Cells with a Gamma Interferon Enzyme-Linked Immunospot Assay

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Published in	Clinical and Vaccine Immunology Vol. 13; no. 6; pp. 684 - 697
Main Authors	Samri, A., Durier, C., Urrutia, A., Sanchez, I., Gahery-Segard, H., Imbart, S., Sinet, M., Tartour, E., Aboulker, J.-P., Autran, B., Venet, A.
Format	Journal Article
Language	English
Published	United States American Society for Microbiology 01.06.2006
Subjects	Antibodies - analysis Antibodies - therapeutic use Cluster Analysis Cytomegalovirus Enzyme-Linked Immunosorbent Assay - methods Epitopes, T-Lymphocyte Epstein-Barr virus HIV Infections - immunology HIV Infections - pathology HIV Infections - therapy HIV Infections - virology Human immunodeficiency virus Humans Interferon-gamma - immunology Peptides - immunology Reproducibility of Results Statistics, Nonparametric T-Lymphocytes - immunology T-Lymphocytes - physiology
Online Access	Get full text
ISSN	1556-6811 1556-679X
DOI	10.1128/CVI.00387-05

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Abstract	Classifications Services CVI Citing Articles Google Scholar PubMed Related Content Social Bookmarking CiteULike Delicious Digg Facebook Google+ Mendeley Reddit StumbleUpon Twitter current issue CVI About CVI Subscribers Authors Reviewers Advertisers Inquiries from the Press Permissions & Commercial Reprints ASM Journals Public Access Policy CVI RSS Feeds 1752 N Street N.W. • Washington DC 20036 202.737.3600 • 202.942.9355 fax • journals@asmusa.org Print ISSN: 1556-6811 Online ISSN: 1556-679X Copyright © 2014 by the American Society for Microbiology. For an alternate route to CVI .asm.org, visit: CVI
AbstractList	The gamma interferon (IFN-gamma) enzyme-linked immunospot (ELISPOT) assay is a reference method for the ex vivo monitoring of antigen-specific T cells and a primary tool for assessing clinical trials of human immunodeficiency virus (HIV) or cancer vaccines. Four experienced laboratories in Paris compared their results with this method by exchanging frozen blood samples from eight HIV-seronegative and eight HIV-seropositive subjects. Each laboratory measured the IFN-gamma-producing cells specific for HIV, Epstein-Barr virus, cytomegalovirus, and influenza using the same set of peptides and the same ELISPOT reader but its own ELISPOT technique. The cutoff values for positive responses (50 or 100 spot-forming cells/10(6) peripheral blood mononuclear cells over background) were consistent with the binomial statistic criterion. The global qualitative concordance, as assessed by the kappa index, ranged from 0.38 to 0.92, that is, moderate to excellent, and was better for non-HIV 9-mer peptide pools than for HIV 15-mer peptide pools. The interlaboratory coefficient of variation for the frequency of virus-specific T cells was 18.7% (data are expressed on a log scale). Clustering analysis of HIV-positive subjects showed qualitative agreement for ELISPOT results from all four laboratories. Overall, the good interlaboratory qualitative concordance of IFN-gamma ELISPOT assays with only the peptide source and ELISPOT reader in common suggests that a qualitative comparison of interlaboratory findings is feasible. Nonetheless, a single set of standard operating procedures should be used in multicenter trials to improve standardization.The gamma interferon (IFN-gamma) enzyme-linked immunospot (ELISPOT) assay is a reference method for the ex vivo monitoring of antigen-specific T cells and a primary tool for assessing clinical trials of human immunodeficiency virus (HIV) or cancer vaccines. Four experienced laboratories in Paris compared their results with this method by exchanging frozen blood samples from eight HIV-seronegative and eight HIV-seropositive subjects. Each laboratory measured the IFN-gamma-producing cells specific for HIV, Epstein-Barr virus, cytomegalovirus, and influenza using the same set of peptides and the same ELISPOT reader but its own ELISPOT technique. The cutoff values for positive responses (50 or 100 spot-forming cells/10(6) peripheral blood mononuclear cells over background) were consistent with the binomial statistic criterion. The global qualitative concordance, as assessed by the kappa index, ranged from 0.38 to 0.92, that is, moderate to excellent, and was better for non-HIV 9-mer peptide pools than for HIV 15-mer peptide pools. The interlaboratory coefficient of variation for the frequency of virus-specific T cells was 18.7% (data are expressed on a log scale). Clustering analysis of HIV-positive subjects showed qualitative agreement for ELISPOT results from all four laboratories. Overall, the good interlaboratory qualitative concordance of IFN-gamma ELISPOT assays with only the peptide source and ELISPOT reader in common suggests that a qualitative comparison of interlaboratory findings is feasible. Nonetheless, a single set of standard operating procedures should be used in multicenter trials to improve standardization. The gamma interferon (IFN-γ) enzyme-linked immunospot (ELISPOT) assay is a reference method for the ex vivo monitoring of antigen-specific T cells and a primary tool for assessing clinical trials of human immunodeficiency virus (HIV) or cancer vaccines. Four experienced laboratories in Paris compared their results with this method by exchanging frozen blood samples from eight HIV-seronegative and eight HIV-seropositive subjects. Each laboratory measured the IFN-γ-producing cells specific for HIV, Epstein-Barr virus, cytomegalovirus, and influenza using the same set of peptides and the same ELISPOT reader but its own ELISPOT technique. The cutoff values for positive responses (50 or 100 spot-forming cells/10 6 peripheral blood mononuclear cells over background) were consistent with the binomial statistic criterion. The global qualitative concordance, as assessed by the kappa index, ranged from 0.38 to 0.92, that is, moderate to excellent, and was better for non-HIV 9-mer peptide pools than for HIV 15-mer peptide pools. The interlaboratory coefficient of variation for the frequency of virus-specific T cells was 18.7% (data are expressed on a log scale). Clustering analysis of HIV-positive subjects showed qualitative agreement for ELISPOT results from all four laboratories. Overall, the good interlaboratory qualitative concordance of IFN-γ ELISPOT assays with only the peptide source and ELISPOT reader in common suggests that a qualitative comparison of interlaboratory findings is feasible. Nonetheless, a single set of standard operating procedures should be used in multicenter trials to improve standardization. Classifications Services CVI Citing Articles Google Scholar PubMed Related Content Social Bookmarking CiteULike Delicious Digg Facebook Google+ Mendeley Reddit StumbleUpon Twitter current issue CVI About CVI Subscribers Authors Reviewers Advertisers Inquiries from the Press Permissions & Commercial Reprints ASM Journals Public Access Policy CVI RSS Feeds 1752 N Street N.W. • Washington DC 20036 202.737.3600 • 202.942.9355 fax • journals@asmusa.org Print ISSN: 1556-6811 Online ISSN: 1556-679X Copyright © 2014 by the American Society for Microbiology. For an alternate route to CVI .asm.org, visit: CVI The gamma interferon (IFN-gamma) enzyme-linked immunospot (ELISPOT) assay is a reference method for the ex vivo monitoring of antigen-specific T cells and a primary tool for assessing clinical trials of human immunodeficiency virus (HIV) or cancer vaccines. Four experienced laboratories in Paris compared their results with this method by exchanging frozen blood samples from eight HIV-seronegative and eight HIV-seropositive subjects. Each laboratory measured the IFN-gamma-producing cells specific for HIV, Epstein-Barr virus, cytomegalovirus, and influenza using the same set of peptides and the same ELISPOT reader but its own ELISPOT technique. The cutoff values for positive responses (50 or 100 spot-forming cells/10(6) peripheral blood mononuclear cells over background) were consistent with the binomial statistic criterion. The global qualitative concordance, as assessed by the kappa index, ranged from 0.38 to 0.92, that is, moderate to excellent, and was better for non-HIV 9-mer peptide pools than for HIV 15-mer peptide pools. The interlaboratory coefficient of variation for the frequency of virus-specific T cells was 18.7% (data are expressed on a log scale). Clustering analysis of HIV-positive subjects showed qualitative agreement for ELISPOT results from all four laboratories. Overall, the good interlaboratory qualitative concordance of IFN-gamma ELISPOT assays with only the peptide source and ELISPOT reader in common suggests that a qualitative comparison of interlaboratory findings is feasible. Nonetheless, a single set of standard operating procedures should be used in multicenter trials to improve standardization. The gamma interferon (IFN- gamma ) enzyme-linked immunospot (ELISPOT) assay is a reference method for the ex vivo monitoring of antigen-specific T cells and a primary tool for assessing clinical trials of human immunodeficiency virus (HIV) or cancer vaccines. Four experienced laboratories in Paris compared their results with this method by exchanging frozen blood samples from eight HIV-seronegative and eight HIV-seropositive subjects. Each laboratory measured the IFN- gamma -producing cells specific for HIV, Epstein-Barr virus, cytomegalovirus, and influenza using the same set of peptides and the same ELISPOT reader but its own ELISPOT technique. The cutoff values for positive responses (50 or 100 spot-forming cells/10 super(6) peripheral blood mononuclear cells over background) were consistent with the binomial statistic criterion. The global qualitative concordance, as assessed by the kappa index, ranged from 0.38 to 0.92, that is, moderate to excellent, and was better for non-HIV 9-mer peptide pools than for HIV 15-mer peptide pools. The interlaboratory coefficient of variation for the frequency of virus-specific T cells was 18.7% (data are expressed on a log scale). Clustering analysis of HIV-positive subjects showed qualitative agreement for ELISPOT results from all four laboratories. Overall, the good interlaboratory qualitative concordance of IFN- gamma ELISPOT assays with only the peptide source and ELISPOT reader in common suggests that a qualitative comparison of interlaboratory findings is feasible. Nonetheless, a single set of standard operating procedures should be used in multicenter trials to improve standardization.
Author	H. Gahery-Segard I. Sanchez A. Samri the ANRS ELISpot Standardization Group B. Autran A. Urrutia S. Imbart J.-P. Aboulker A. Venet C. Durier E. Tartour M. Sinet
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et Marie Curie—Paris 6, Paris, France, INSERM SC10, Villejuif, FranceLaboratoire d’Immunité Antivirale, AP-HP, Hôpital Kremlin-Bicêtre,, Faculté de Médicine Paris-Sud, Université Paris XI, Kremlin-Bicêtre, France, INSERM U567, Hôpital Cochin, Paris, France, Unité d'Immunologie Biologique Hôpital AP-HP Européen Georges Pompidou, UMR S 255, Université Paris V René Descartes, Paris, France – sequence: 5 givenname: H. surname: Gahery-Segard fullname: Gahery-Segard, H. organization: Laboratoire d'Immunologie Cellulaire, AP-HP, Hôpital Pitié-Salpêtrière, INSERM UMR S 543, Université Pierre et Marie Curie—Paris 6, Paris, France, INSERM SC10, Villejuif, FranceLaboratoire d’Immunité Antivirale, AP-HP, Hôpital Kremlin-Bicêtre,, Faculté de Médicine Paris-Sud, Université Paris XI, Kremlin-Bicêtre, France, INSERM U567, Hôpital Cochin, Paris, France, Unité d'Immunologie Biologique Hôpital AP-HP Européen Georges Pompidou, UMR S 255, Université Paris V René Descartes, 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Kremlin-Bicêtre, France, INSERM U567, Hôpital Cochin, Paris, France, Unité d'Immunologie Biologique Hôpital AP-HP Européen Georges Pompidou, UMR S 255, Université Paris V René Descartes, Paris, France – sequence: 8 givenname: E. surname: Tartour fullname: Tartour, E. organization: Laboratoire d'Immunologie Cellulaire, AP-HP, Hôpital Pitié-Salpêtrière, INSERM UMR S 543, Université Pierre et Marie Curie—Paris 6, Paris, France, INSERM SC10, Villejuif, FranceLaboratoire d’Immunité Antivirale, AP-HP, Hôpital Kremlin-Bicêtre,, Faculté de Médicine Paris-Sud, Université Paris XI, Kremlin-Bicêtre, France, INSERM U567, Hôpital Cochin, Paris, France, Unité d'Immunologie Biologique Hôpital AP-HP Européen Georges Pompidou, UMR S 255, Université Paris V René Descartes, Paris, France – sequence: 9 givenname: J.-P. surname: Aboulker fullname: Aboulker, J.-P. organization: Laboratoire d'Immunologie Cellulaire, AP-HP, Hôpital Pitié-Salpêtrière, INSERM UMR S 543, Université 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Snippet	Classifications Services CVI Citing Articles Google Scholar PubMed Related Content Social Bookmarking CiteULike Delicious Digg Facebook Google+ Mendeley Reddit... The gamma interferon (IFN-γ) enzyme-linked immunospot (ELISPOT) assay is a reference method for the ex vivo monitoring of antigen-specific T cells and a... The gamma interferon (IFN-gamma) enzyme-linked immunospot (ELISPOT) assay is a reference method for the ex vivo monitoring of antigen-specific T cells and a... The gamma interferon (IFN- gamma ) enzyme-linked immunospot (ELISPOT) assay is a reference method for the ex vivo monitoring of antigen-specific T cells and a...
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SubjectTerms	Antibodies - analysis Antibodies - therapeutic use Cluster Analysis Cytomegalovirus Enzyme-Linked Immunosorbent Assay - methods Epitopes, T-Lymphocyte Epstein-Barr virus HIV Infections - immunology HIV Infections - pathology HIV Infections - therapy HIV Infections - virology Human immunodeficiency virus Humans Interferon-gamma - immunology Peptides - immunology Reproducibility of Results Statistics, Nonparametric T-Lymphocytes - immunology T-Lymphocytes - physiology
Title	Evaluation of the Interlaboratory Concordance in Quantification of Human Immunodeficiency Virus-Specific T Cells with a Gamma Interferon Enzyme-Linked Immunospot Assay
URI	http://cvi.asm.org/content/13/6/684.abstract https://www.ncbi.nlm.nih.gov/pubmed/16760328 https://www.proquest.com/docview/17234572 https://www.proquest.com/docview/68045033
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