Evaluation of antigen-specific T-cell responses with a miniaturized and automated method

Giuseppina Li Pira, Federico Ivaldi, Chiara Dentone, Elda Righi, Valerio Del Bono, Claudio Viscoli, Gerrit Koopman, Fabrizio Manca

Research output: Contribution to journalArticle

Abstract

The evaluation of antigen-specific T-cell responses is helpful for both research and clinical settings. Several techniques can enumerate antigen-responsive T cells or measure their products, but they require remarkable amounts of peripheral blood mononuclear cells (PBMCs). Since screening numerous antigens or testing samples from pediatric or lymphopenic patients is hampered in clinical practice, we refined a miniaturized, high-throughput assay for T-cell immunity. Antigens and cells in 10-μl volumes were dispensed into 1,536-well culture plates precoated with anti-gamma interferon (anti-IFN-γ) antibodies. After being cultured, the wells were developed by enzyme-linked immunosorbent assay for bound cytokine. Miniaturization and automation allowed quantitation of antigen-specific responses on 104 PBMCs. This method was applied for epitope mapping of mycobacterial antigens and was used in the clinic to evaluate T-cell immunity to relevant opportunistic pathogens by using small blood samples. A comparison with conventional methods showed similar sensitivity. Therefore, current flow cytometric methods that provide information on frequency and phenotype of specific T cells can be complemented by this assay that provides extensive information on cytokine concentrations and profiles and requires 20- to 50-fold fewer PBMCs than other analytical methods.

Original languageEnglish
Pages (from-to)1811-1818
Number of pages8
JournalClinical and Vaccine Immunology
Volume15
Issue number12
DOIs
Publication statusPublished - Dec 2008

ASJC Scopus subject areas

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

Fingerprint Dive into the research topics of 'Evaluation of antigen-specific T-cell responses with a miniaturized and automated method'. Together they form a unique fingerprint.

  • Cite this

    Li Pira, G., Ivaldi, F., Dentone, C., Righi, E., Del Bono, V., Viscoli, C., Koopman, G., & Manca, F. (2008). Evaluation of antigen-specific T-cell responses with a miniaturized and automated method. Clinical and Vaccine Immunology, 15(12), 1811-1818. https://doi.org/10.1128/CVI.00322-08