Evaluation of T Cell Proliferation Using CFSE Dilution Assay: A Comparison between Stimulation with PHA and Anti-CD3/Anti-CD28 Coated Beads

Sara Kashef; Mozhgan Moghtaderi; Hamid Reza Hatami; Mehdi Kalani; Soheila Alyasin; Hesamodin Nabavizadeh; Shirin Farjadian

doi:10.18502/ijaai.v21i4.10293

Sara Kashef Department of Allergy and Clinical Immunology, Shiraz University of Medical Sciences, Shiraz, Iran
Mozhgan Moghtaderi Allergy Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
Hamid Reza Hatami Department of Allergy and Clinical Immunology, Shiraz University of Medical Sciences, Shiraz, Iran
Mehdi Kalani Prof. Alborzi Clinical Microbiology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
Soheila Alyasin Department of Allergy and Clinical Immunology, Shiraz University of Medical Sciences, Shiraz, Iran
Hesamodin Nabavizadeh Department of Allergy and Clinical Immunology, Shiraz University of Medical Sciences, Shiraz, Iran
Shirin Farjadian Department of Immunology, Shiraz University of Medical Sciences, Shiraz, Iran

DOI: https://doi.org/10.18502/ijaai.v21i4.10293

Keywords: Anti-CD3; Anti-CD28; Carboxyfluorescein diacetate succinimidyl ester; Lymphocyte transformation; Phytohaemagglutinin

Abstract

A decrease in T cell count or reduced T cell function can be indicative of T cell immunodeficiency. In the present study, T-cell function was assessed using Carboxyfluorescein diacetate succinimidyl ester (CFSE) dilution test after stimulation with commonly used Phytohaemagglutinin (PHA) or anti-CD3/anti-CD28 coated beads in pediatric patients with recurrent infections.

Seven infants with recurrent infections and seven sex/age-matched healthy infants were included in this study. A blood cell count, immunophenotyping, and serum immunoglobulin level were performed. The proliferation of T cells was also assessed with CFSE dilution after stimulation with PHA or anti-CD3/anti-CD28 coated beads.

This study showed increased IgA, IgG, and IgM levels in patients compared to the controls. In contrast to the controls, the immunophenotyping results showed a significant decline in the number of CD4⁺ T cells in patients. Although there was no difference in CD3⁺ T cell proliferation between patients and controls, the CD4⁺ and CD8⁺ T cell proliferation rates were significantly decreased in patients when stimulated with PHA.

As a mitogen with the potential for maximum proliferation of T cells, PHA is better able to distinguish between patients with recurrent infections and controls than anti-CD3/anti-CD28, which mimics only the TCR pathway for stimulation of T cells.