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An accurate and rapid single step protocol for enumeration of cytokine positive T lymphocytes



Deepa Rajagopal, Linhua Tian, Shiqiu Xiong, Lili Wang, Jonathan Campbell, Luisa Saraiva, S Vessillier


Activation of T lymphocytes leads to differentiation, characterized by the ability to induce production of cytokines. Accurate determination of cellular subsets that secrete particular cytokine(s) is therefore, a significant parameter for functional characterization of an immunological response. The present study was conducted to develop a method to simultaneously measure intracellular cytokine positive CD4 and CD8 positive T cell numbers in a single tube with a no-wash protocol. We report here the development of a simplified, rapid procedure for accurate enumeration cytokine positive cells using BD Trucount tubes. This single step protocol allows for accurate enumeration and characterization of immune cell phenotype.
Journal of Immunology and Regenerative Medicine


Cytokine positive T lymphocytes, enumeration, flow cytometry, single step protocol, two step protocol, Trucount bead standard, isotype control, INF-r


Rajagopal, D. , Tian, L. , Xiong, S. , Wang, L. , Campbell, J. , Saraiva, L. and Vessillier, S. (2020), An accurate and rapid single step protocol for enumeration of cytokine positive T lymphocytes, Journal of Immunology and Regenerative Medicine, [online], (Accessed May 17, 2024)


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Created September 2, 2020, Updated October 12, 2021