Comparison of immunologic assays for detecting immune responses in HIV immunotherapeutic studies

AIDS clinical trials group trial A5181

Bernard J C Macatangay, Lu Zheng, Charles R. Rinaldo, Alan L. Landay, Richard B Pollard, Savita Pahwa, Michael M. Lederman, R. Pat Bucy

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

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-γ) cytokine production (CP), single-cell IFN-γ 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-γ 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-γ 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-γ 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-γ-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.

Original languageEnglish (US)
Pages (from-to)1452-1459
Number of pages8
JournalClinical and Vaccine Immunology
Volume17
Issue number9
DOIs
StatePublished - Sep 2010

Fingerprint

Enzyme-Linked Immunospot Assay
Interferons
HIV-1
Assays
Acquired Immunodeficiency Syndrome
Clinical Trials
HIV
Cytokines
HIV Antigens
T-cells
T-Lymphocytes
Interleukin-2
Enzymes
Lymphocytes
Immunologic Monitoring
Cell Line
Antigens
Viremia
Group Psychotherapy
Human Herpesvirus 4

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Immunology
  • Immunology and Allergy
  • Microbiology (medical)

Cite this

Comparison of immunologic assays for detecting immune responses in HIV immunotherapeutic studies : AIDS clinical trials group trial A5181. / Macatangay, Bernard J C; Zheng, Lu; Rinaldo, Charles R.; Landay, Alan L.; Pollard, Richard B; Pahwa, Savita; Lederman, Michael M.; Bucy, R. Pat.

In: Clinical and Vaccine Immunology, Vol. 17, No. 9, 09.2010, p. 1452-1459.

Research output: Contribution to journalArticle

Macatangay, Bernard J C ; Zheng, Lu ; Rinaldo, Charles R. ; Landay, Alan L. ; Pollard, Richard B ; Pahwa, Savita ; Lederman, Michael M. ; Bucy, R. Pat. / Comparison of immunologic assays for detecting immune responses in HIV immunotherapeutic studies : AIDS clinical trials group trial A5181. In: Clinical and Vaccine Immunology. 2010 ; Vol. 17, No. 9. pp. 1452-1459.
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