 |
|
- Main page
- News
- RJLabM Credits
- Archive
- Instructions for authors
- Online applications
- Editorial
- About us
-
Links
- Committee on Publication Ethics (COPE)
- International Committee of Medical Journal Editors
- Acta Marisiensis Seria Medica
- Bacteriologia, Virusologia, Parazitologia, Epidemiologia
- Biochemia Medica
- Clinical Chemistry and Laboratory Medicine
- Clinical Chemistry
- Clinica Chimica Acta
- Documenta Haematologica
- Romanian Journal of Morphology and Embryology
- Biostatistics and web search strategies (Romanian)
- How to spot a statistical problem
- Statistical Analyses and Methods in the Published Literature
ISSN online: 2284-5623
ISSN-L: 1841-6624
Rejection rate (2020): 75%
Journal Metrics
Impact Factor 0.5 Five Year Impact Factor 0.5 JCI 0.12
Log in
Concept, Design & Programming
Dr. Adrian Man
Short communication Variability of ex-vivo stimulated T-cells secretory profile in healthy subjects Ion Bogdan Manescu, Doina Ramona Manu, Georgiana Mihaela Serban, Minodora Dobreanu
Correspondence should be addressed to: Doina Ramona Manu
Abstract: Peripheral blood lymphocytes (PBL) are able to synthesize various cytokines that play key roles in the immune response and intercellular signaling. Since alterations in cytokine production and/or activity occur in many pathological processes, the study of cytokine synthetic capacity of PBL is a valuable tool for assessing the immune profile. In this paper, we aimed to investigate the variability of interleukin-2 (IL-2), tumor necrosis factor-alpha (TNF-α) and interferon gamma (IFN-γ) synthetic capacity of CD4+/CD8+ T-cells stimulated ex-vivo in healthy subjects, by means of a commercial intracellular cytokine staining (ICS) protocol. Peripheral blood mononuclear cells were isolated from 16 healthy subjects by Ficoll gradient centrifugation and activated ex-vivo with PMA/Ionomycin/Brefeldin-A for 4 hours. Activated PBL were surface-stained for CD3/CD4/CD8, fixed and permeabilized. ICS was performed using anti-human IL-2/TNF-α/IFN-γ and samples were analyzed on a BD-FACSAria-III flow cytometer. We recorded high post-isolation and post-activation mean viabilities: 82.1% and 82.4% respectively, p=0.84. Both CD4+/CD8+ subpopulations were found to partially produce each of the three cytokines, but in different proportions. On average, a significantly greater percentage of CD4+ cells was shown to produce IL-2 and TNF-α, compared with CD8+ cells (61.5%+/-5.8 vs. 25%+/-5.6 and 26.9%+/-11 vs. 7.5%+/-3.3 respectively, p<0.0001 for both). Contrarily, IFN-γ was produced by a higher proportion of CD8+ cells (8.4%+/-3.9 vs. 6.8%+/-3.2, p=0.01). These results show that the employed ICS protocol elicits a satisfactory and consistent cytokine response from PBL of healthy subjects. The collected data may be used to outline a preliminary reference range for future studies on both healthy/pathological subjects. Keywords: peripheral blood lymphocytes, ex-vivo T-cells stimulation, cytokines, flow cytometry, reference intervals
Received: 2.10.2019 |
|||||
|
Download full text PDF (898 KB) |
||||
|
Manescu IB, Manu DR, Serban GM, Dobreanu M. Variability of ex-vivo stimulated T-cells secretory profile in healthy subjects. Rev Romana Med Lab. 2020;28(1):75-89. DOI:10.2478/rrlm-2020-0004
|
|||||