Concordant Proficiency in Measurement of T-Cell Immunity in Human Immunodeficiency Virus Vaccine Clinical Trials by Peripheral Blood Mononuclear Cell and Enzyme-Linked Immunospot Assays in Laboratories from Three Continents

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dc.contributor.authorBoaz, Mark J.
dc.contributor.authorHayes, Peter
dc.contributor.authorTarragona, Tony
dc.contributor.authorSeamons, Laura
dc.contributor.authorCooper, Andrew
dc.contributor.authorBirungi, Josephine
dc.contributor.authorKitandwe, Paul
dc.contributor.authorSemaganda, Aloysius
dc.contributor.authorKaleebu, Pontiano
dc.contributor.authorStevens, Gwynneth
dc.contributor.authorAnzala, Omu
dc.contributor.authorFarah, Bashir
dc.contributor.authorOgola, Simon
dc.contributor.authorIndangasi, Jackton
dc.contributor.authorMhlanga, Patrick
dc.contributor.authorEeden, Melanie Van
dc.contributor.authorThakar, Madhuri
dc.contributor.authorPujari, Ashwini
dc.contributor.authorMishra, Shadri
dc.contributor.authorGoonetilleke, Nilu
dc.contributor.authorMoore, Stephen
dc.contributor.authorMahmoud, Abdul
dc.contributor.authorSathyamoorthy, Pattabiraman
dc.contributor.authorMahalingam, Jayashri
dc.contributor.authorNarayanan, Paranji R.
dc.contributor.authorRamanathan, Vadakkuppattu D.
dc.contributor.authorCox, Josephine H.
dc.contributor.authorDally, Len
dc.contributor.authorGill, Dilbinder K.
dc.contributor.authorGilmour, Jill
dc.date.accessioned2018-08-06T08:18:20Z
dc.date.available2018-08-06T08:18:20Z
dc.date.issued2009-02
dc.descriptionThis study was focused on the gamma interferon (IFN-_) enzyme-linked immunospot (ELISPOT) assay used routinely to evaluate the potency of human immunodeficiency virus (HIV) vaccine candidates and other vaccine candidatesen_US
dc.description.abstractThe gamma interferon (IFN-_) enzyme-linked immunospot (ELISPOT) assay is used routinely to evaluate the potency of human immunodeficiency virus (HIV) vaccine candidates and other vaccine candidates. In order to compare candidates and pool data from multiple trial laboratories, validated standardized methods must be applied across laboratories. Proficiency panels are a key part of a comprehensive quality assurance program to monitor inter- and intralaboratory performance, as well as assay performance, over time. Seven International AIDS Vaccine Initiative-sponsored trial sites participated in the proficiency panels described in this study. At each laboratory, two operators independently processed identical sample sets consisting of frozen peripheral blood mononuclear cell (PBMC) samples from different donors by using four blind stimuli. PBMCM recovery and viability after overnight resting and the IFN-_ ELISPOT assay performance were assessed. All sites demonstrated good performance in PBMC thawing and resting, with a median recovery of 78% and median viability of 95%. The laboratories were able to detect similar antigen-specific T-cell responses, ranging from 50 to >3,000 spot-forming cells per million PBMC. An approximate range of a half log in results from operators within or across sites was seen in comparisons of antigen-specific responses. Consistently low background responses were seen in all laboratories. The results of these proficiency panels demonstrate the ability of seven laboratories, located across three continents, to process PBMC samples and to rank volunteers with differential magnitudes of IFN-_ ELISPOT responses. These findings also illustrate the ability to standardize the IFN-_ ELISPOT assay across multiple laboratories when common training methods, reagents such as fetal calf serum, and standard operating procedures are adopted. These results are encouraging for laboratories that are using cell-based immunology assays to test HIV vaccines and other vaccines.en_US
dc.identifier.citationBoaz et al. Concordant Proficiency in Measurement of T-Cell Immunity in Human Immunodeficiency Virus Vaccine Clinical Trials by Peripheral Blood Mononuclear Cell and Enzyme-Linked Immunospot Assays in Laboratories from Three Continents. Clinical and vaccine immunology, Vol. 16 No. 2 p. 147–155. 1556. (Feb. 2009) DOI:10.1128/CVI.00326-08en_US
dc.identifier.urihttps://hdl.handle.net/20.500.11951/326
dc.language.isoenen_US
dc.publisherClinical and vaccine immunologyen_US
dc.subjectT-Cell Immunityen_US
dc.subjectHuman Immunodeficiencyen_US
dc.subjectVirus Vaccine Clinical Trialsen_US
dc.subjectPeripheral Blooden_US
dc.subjectMononuclear Cellen_US
dc.subjectEnzyme-Linked Immunospoten_US
dc.titleConcordant Proficiency in Measurement of T-Cell Immunity in Human Immunodeficiency Virus Vaccine Clinical Trials by Peripheral Blood Mononuclear Cell and Enzyme-Linked Immunospot Assays in Laboratories from Three Continentsen_US
dc.typeArticleen_US
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The final, definitive version of this paper has been published in the Clinical and vaccine immunology, Feb. 2009, Vol. 16 No. 2 p. 147–155. 1556. DOI:10.1128/CVI.00326-08, published by the American Society for Microbiology. All rights reserved.
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