Equivalence of ELISpot assays demonstrated between major HIV network laboratories

The Comprehensive T Cell Vaccine Immune Monitoring Consortium (CTC-VIMC) was created to provide standardized immunogenicity monitoring services for HIV vaccine trials. The ex vivo interferon-gamma (IFN-γ) ELISpot is used extensively as a primary immunogenicity assay to assess T cell-based vaccine ca...

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Bibliographic Details
Published in:PloS one 2010-12, Vol.5 (12), p.e14330-e14330
Main Authors: Gill, Dilbinder K, Huang, Yunda, Levine, Gail L, Sambor, Anna, Carter, Donald K, Sato, Alicia, Kopycinski, Jakub, Hayes, Peter, Hahn, Bridget, Birungi, Josephine, Tarragona-Fiol, Tony, Wan, Hong, Randles, Mark, Cooper, Andrew Raxworthy, Ssemaganda, Aloysius, Clark, Lorna, Kaleebu, Pontiano, Self, Steven G, Koup, Richard, Wood, Blake, McElrath, M Juliana, Cox, Josephine H, Hural, John, Gilmour, Jill
Format: Article
Language:English
Subjects:
Acceptance criteria
Acceptance tests
Acquired immune deficiency syndrome
AIDS
AIDS Serodiagnosis - methods
AIDS Serodiagnosis - standards
AIDS vaccines
AIDS Vaccines - therapeutic use
Analysis
Antigens
Biological response modifiers
Cancer
Clinical trials
Communicable diseases
Confidence intervals
Consortia
Data collection
Data processing
Enzyme-linked immunosorbent assay
Enzyme-Linked Immunosorbent Assay - methods
Enzymes
Equivalence
Gangrene
HIV
HIV Infections - prevention & control
HIV Seronegativity
HIV tests
HIV-1 - metabolism
Human immunodeficiency virus
Humans
Immune System
Immunogenicity
Immunology
Infectious Diseases
Infectious Diseases/HIV Infection and AIDS
Infectious Diseases/Sexually Transmitted Diseases
Infectious Diseases/Viral Infections
Interferon
Interferon-gamma - metabolism
Laboratories
Laboratories - standards
Leukocytes, Mononuclear - metabolism
Lymphocytes
Lymphocytes T
Medical research
Models, Statistical
Monitoring
Peptides
Pilot Projects
Prevention
Quality control
Reproducibility of Results
Statistical analysis
T cells
T-Lymphocytes - metabolism
Testing laboratories
Tumor necrosis factor-TNF
Vaccines
γ-Interferon
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Summary:The Comprehensive T Cell Vaccine Immune Monitoring Consortium (CTC-VIMC) was created to provide standardized immunogenicity monitoring services for HIV vaccine trials. The ex vivo interferon-gamma (IFN-γ) ELISpot is used extensively as a primary immunogenicity assay to assess T cell-based vaccine candidates in trials for infectious diseases and cancer. Two independent, GCLP-accredited central laboratories of CTC-VIMC routinely use their own standard operating procedures (SOPs) for ELISpot within two major networks of HIV vaccine trials. Studies are imperatively needed to assess the comparability of ELISpot measurements across laboratories to benefit optimal advancement of vaccine candidates. We describe an equivalence study of the two independently qualified IFN-g ELISpot SOPs. The study design, data collection and subsequent analysis were managed by independent statisticians to avoid subjectivity. The equivalence of both response rates and positivity calls to a given stimulus was assessed based on pre-specified acceptance criteria derived from a separate pilot study. Detection of positive responses was found to be equivalent between both laboratories. The 95% C.I. on the difference in response rates, for CMV (-1.5%, 1.5%) and CEF (-0.4%, 7.8%) responses, were both contained in the pre-specified equivalence margin of interval [-15%, 15%]. The lower bound of the 95% C.I. on the proportion of concordant positivity calls for CMV (97.2%) and CEF (89.5%) were both greater than the pre-specified margin of 70%. A third CTC-VIMC central laboratory already using one of the two SOPs also showed comparability when tested in a smaller sub-study. The described study procedure provides a prototypical example for the comparison of bioanalytical methods in HIV vaccine and other disease fields. This study also provides valuable and unprecedented information for future vaccine candidate evaluations on the comparison and pooling of ELISpot results generated by the CTC-VIMC central core laboratories.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0014330