Determinación de poblaciones circulantes en sangre periférica de linfocitos T, reguladores CD4+ CD25+ CD127-, en niños asmáticos alérgicos

Abstract

Introduction: The type 2 immune response participates in the defense against parasitic infections, but in the context of asthma and allergies, this response can react disproportionately to environmental allergens, triggering inflammation and airway hyperreactivity. Regulatory T cells (Tregs) play a fundamental role in modulating and controlling the inflammatory response; thereby, a decrease or dysfunction of Tregs could result in increased sensitization to allergens, aggravating asthma and allergies. Evaluations of Tregs usually include intracellular stains for Foxp3, but a new immunophenotyping panel can detect Tregs by staining with CD25 and CD127.

Objective: To carry out a preliminary analysis on the Treg lymphocyte counts present in the peripheral blood of allergic asthmatic children from the city of Cartagena (Colombia) compared to healthy controls.

Methodology: We compared cytometry counts of 10 asthmatic patients (age 7-16 years) and 7 healthy controls (6-12 years) recruited in the city of Cartagena. Peripheral blood samples were stained using Cytek’s 14-color cFluor Immunoprofiling kit (Cytek® cFluor® Immunoprofiling Kit 14 Color RUO kit) and analyzed on a Northern Lights™ spectral cytometer (Cytek® Biosciences, Fremont, CA, USA) to read 50,000 events per sample. The data obtained were analyzed in SpectroFlo® and FlowJo. The study was approved by the ethics committee of the University of Cartagena (SGR, Grant BPIN2020000100405).

Results: The frequency of CD3+ CD4+ CD25+ CD127- Tregs was 11% of all CD4+ T cells, with a range of minimum 8.1% and maximum 17.7%. There was no significant difference in the proportion of Tregs between allergic asthmatic patients and healthy controls (P = 0.2).

Conclusions: With this preliminary sample size, no significant differences were found in the Treg lymphocyte population between allergic asthmatic patients and healthy controls. The 14-color multiplexed panel is a useful tool not only to count CD3+ and CD4+ populations but also to obtain the percentage of regulatory T cells using cell surface markers.

Funding: This study was funded by the SGR, project BPIN2020000100405 and the University of Cartagena.

Declaration of conflict of interest: The authors declare that they have no conflicts of interest in relation to this work.