Use of a short-term whole blood intracellular staining assay to study the T-cell response in respiratory syncytial virus-infected pediatric patients

A. Frassanito, G. Fedele, P. Leone, R. Nenna, F. Midulla, Ilaria Schiavoni

Research output: Contribution to journalLetterpeer-review

Abstract

The aim of the present study was the development of a reliable method to evaluate the pattern of the ongoing T-cell response in young infants affected by respiratory infection. To this purpose, we enrolled 44 infants hospitalized with a diagnosis of respiratory syncytial virus bronchiolitis. After a short-term stimulation of whole blood samples, intracellular IFN-γ and IL-4 cytokines were measured in CD4 + and CD8 + T-cell subsets by flow cytometry. A stringent staining and gating strategy was used in order to maximize the reduction of background noise and to exclude false positives. The frequencies of cytokine-producing T-cell subsets, albeit low, were easily quantifiable. Cytokine responses were higher in infants sampled > 7 days from the onset of symptoms. The use of a rigorous strategy for cell staining and gating, coupled with a short-term stimulation of whole blood and a careful evaluation of time elapsed from the onset of symptoms constitutes a convincing approach for future clinical studies.

Original languageEnglish
Pages (from-to)1339-1344
Number of pages6
JournalJournal of Biological Regulators and Homeostatic Agents
Volume32
Issue number5
Publication statusPublished - Sep 1 2018

Keywords

  • Bronchiolitis
  • Cytokines
  • Flow-cytometry
  • Respiratory syncytial virus
  • T-cell

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Immunology and Allergy
  • Physiology
  • Immunology
  • Oncology
  • Endocrinology
  • Physiology (medical)
  • Cancer Research

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