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Comparison of mitogen-induced proliferation in child and adult healthy groups by flow cytometry revealed similarities

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Abstract

The proliferation of antigen-specific lymphocyte clones, the initial step in acquired immunity, is vital for effector functions. Proliferation tests both in immunology research and diagnosis are gaining attendance gradually, while the use of adult healthy individuals as controls of pediatric patients is a question. This study aimed to investigate and compare mitogen-stimulated proliferation responses of total lymphocytes and T- and B-lymphocyte subsets in adult and children healthy donors. Nineteen children and 20 adult healthy donors were enrolled in this study. Peripheral blood mononuclear cells (PBMCs) purified from peripheral blood samples of the donors, by Ficoll gradient centrifugation, were stained with CFSE and were cultured in a 37 ℃ CO2 incubator for 120 h with the absence or existence of polyclonal activators: PHA and CD-Mix. After cell culture, PBMCs were stained with monoclonal antibodies against CD4 and CD19, and proliferation percentages of CD4+ T and CD19+ B cells, together with total lymphocytes were determined by flow cytometry. This study revealed similarities between children and adult age groups, concerning mitogenic stimulation of the lymphocytes. The only difference was a significantly high proliferation of pediatric CD4+ T cells in response to PHA. CD4+ T cell responses against PHA were inversely correlated with altering age. When pediatric individuals were distributed into age groups of 0–2 years, 3–5 years, and 6–18 years, PHA responses of CD4+ cells were found to be diminished with advancing age. These findings propose the possibility of enrollment of adult healthy individuals as controls for pediatric patients.

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Data availability

The data related with this study is available upon request from the corresponding author.

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Acknowledgements

The authors like to thank Mr. Abdullah Yilmaz for his important assistance during the laboratory works.

Funding

This present work was supported by the Research Fund of Istanbul University. Project No. 29901.

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Authors and Affiliations

  1. Graduate School of Health Sciences, Istanbul University, Vakif Gureba St. 34093, Fatih, Istanbul, Turkey

    Zakya Shoub Elshari

  2. Department of Immunology, Aziz Sancar Institute of Experimental Medicine, Istanbul University, Vakif Gureba St. 34093, Fatih, Istanbul, Turkey

    Zakya Shoub Elshari, Ilhan Tahrali, Gunnur Deniz & Umut Can Kucuksezer

  3. Department of Pediatrics, Division of Pediatric Allergy and Immunology, Cerrahpasa Faculty of Medicine, Istanbul University-Cerrahpasa, Istanbul, Turkey

    Serdar Nepesov, Ayca Kiykim & Yildiz Camcioglu

  4. Department of Pediatrics, Division of Pediatric Allergy and Immunology, Medical Park Goztepe Hospital, Istanbul, Turkey

    Serdar Nepesov

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  1. Zakya Shoub Elshari
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Contributions

All authors contributed to the study conception and design. Zakya Shoub Elshari carried out the experiments, collected the data, and contributed in the initial draft of the manuscript. Serdar Nepesov, Ayca Kiykim, and Yildiz Camcioglu coordinated the establishment of donor groups and obtained the blood samples and also contributed in finalization of the manuscript. Ilhan Tahrali contributed in experiments, data analysis, and construction of graphics. Gunnur Deniz contributed in experiment design, discussion of data, and also proofreading. Umut Can Kucuksezer contributed in acquisition of adult samples, design and conduction of experiments, statistical analysis, graphics design, finalization of the manuscript, and proofreading.

Corresponding author

Correspondence to Umut Can Kucuksezer.

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Ethics approval

This study was performed in line with the principles of the Declaration of Helsinki. Approval was granted by the local ethical committee of Istanbul University, Istanbul Faculty of Medicine (date: 23–06-2017/ no:817). The written informed consents for participation in this study were obtained from all of the participants or from their parents.

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Elshari, Z.S., Nepesov, S., Tahrali, I. et al. Comparison of mitogen-induced proliferation in child and adult healthy groups by flow cytometry revealed similarities. Immunol Res 71, 51–59 (2023). https://doi.org/10.1007/s12026-022-09328-2

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  • DOI: https://doi.org/10.1007/s12026-022-09328-2

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