Document Detail


Comparison of immunologic assays for detecting immune responses in HIV immunotherapeutic studies: AIDS Clinical Trials Group Trial A5181.
MedLine Citation:
PMID:  20631337     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
This study was designed to evaluate which of several T-cell-specific, immune response assays are the most relevant in measuring the key characteristics of an effective immune response to HIV-1. Using 5 HIV-1 antigens as stimulants, we assessed lymphocyte proliferation, supernatant gamma interferon (IFN-gamma) cytokine production (CP), single-cell IFN-gamma production by enzyme-linked immunospot (ELISPOT) assay, with and without Epstein-Barr virus-transformed B-lymphoblastoid cell lines (B-LCLs), and intracellular cytokine production (ICC) for IFN-gamma and interleukin 2 (IL-2) by flow cytometry. We used these to compare specimens from HIV-1-infected subjects who were virally suppressed with a stable antiretroviral therapy (ART) regimen (group A) with specimens from subjects not on ART but with HIV-1 viremia of <3,000 copies/ml (group B). The lymphocyte proliferation assay (LPA) did not significantly differentiate between the two groups. Using fresh peripheral blood mononuclear cells (PBMCs), the CP and ELISPOT assays for IFN-gamma detected the greatest differences between the two groups, specific for three of the five HIV-1 antigens, whereas significant differences were seen only in response to one antigen when cryopreserved cells were used. The strongest correlations were seen between the CP and ELISPOT assays. The ELISPOT B-LCL assay showed a cell concentration-dependent increase in IFN-gamma production compared to that shown by the standard ELISPOT assay but did not differentiate between the groups. In the ICC assay, greater numbers of IFN-gamma-producing T cells were seen in group B, and little or no detectable IL-2 production was seen in both groups. These studies highlight complexities of immunologic monitoring of T-cell responses in multisite clinical trials in HIV infection and outline considerations for optimizing these efforts.
Authors:
Bernard J C Macatangay; Lu Zheng; Charles R Rinaldo; Alan L Landay; Richard B Pollard; Savita Pahwa; Michael M Lederman; R Pat Bucy
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Publication Detail:
Type:  Comparative Study; Evaluation Studies; Journal Article; Research Support, N.I.H., Extramural     Date:  2010-07-14
Journal Detail:
Title:  Clinical and vaccine immunology : CVI     Volume:  17     ISSN:  1556-679X     ISO Abbreviation:  Clin. Vaccine Immunol.     Publication Date:  2010 Sep 
Date Detail:
Created Date:  2010-09-02     Completed Date:  2010-12-08     Revised Date:  2011-07-25    
Medline Journal Info:
Nlm Unique ID:  101252125     Medline TA:  Clin Vaccine Immunol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1452-9     Citation Subset:  IM    
Affiliation:
Division of Infectious Diseases/HIV/AIDS Unit, University of Pittsburgh School of Medicine, Pittsburgh, PA 15213, USA. macatangaybj@upmc.edu
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MeSH Terms
Descriptor/Qualifier:
Adolescent
Adult
Aged
Anti-HIV Agents / therapeutic use
Antigens, Viral / diagnostic use
Antiretroviral Therapy, Highly Active / methods
Cell Proliferation
Female
Flow Cytometry
HIV Infections / drug therapy,  immunology*,  therapy*
HIV-1 / immunology*
Humans
Immunoassay / methods
Immunotherapy / methods*
Interferon-gamma / biosynthesis,  secretion
Interleukin-2 / biosynthesis
Leukocytes, Mononuclear / immunology
Male
Middle Aged
Virology / methods*
Young Adult
Grant Support
ID/Acronym/Agency:
AI038858/AI/NIAID NIH HHS; AI069452/AI/NIAID NIH HHS; AI069471/AI/NIAID NIH HHS; AI27675/AI/NIAID NIH HHS; AI54907/AI/NIAID NIH HHS; AI68634/AI/NIAID NIH HHS; AI68638/AI/NIAID NIH HHS; AI69450/AI/NIAID NIH HHS; AI69477/AI/NIAID NIH HHS; AI69494/AI/NIAID NIH HHS; AI69501/AI/NIAID NIH HHS; AI73961/AI/NIAID NIH HHS; RR25780/RR/NCRR NIH HHS; U01AI068636/AI/NIAID NIH HHS
Chemical
Reg. No./Substance:
0/Anti-HIV Agents; 0/Antigens, Viral; 0/Interleukin-2; 82115-62-6/Interferon-gamma
Comments/Corrections

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