Cells 2017, 6(3), 29; https://doi.org/10.3390/cells6030029
Delayed Activation Kinetics of Th2- and Th17 Cells Compared to Th1 Cells
1
R & D Department CTL, Shaker Hts, OH 44122, USA
2
Department Anatomy, University Erlangen, 91054 Erlangen, Germany
*
Author to whom correspondence should be addressed.
Received: 5 August 2017 / Revised: 29 August 2017 / Accepted: 5 September 2017 / Published: 12 September 2017
(This article belongs to the Special Issue Recent Advances in ELISPOT Research)
Abstract
During immune responses, different classes of T cells arise: Th1, Th2, and Th17. Mobilizing the right class plays a critical role in successful host defense and therefore defining the ratios of Th1/Th2/Th17 cells within the antigen-specific T cell repertoire is critical for immune monitoring purposes. Antigen-specific Th1, Th2, and Th17 cells can be detected by challenging peripheral blood mononuclear cells (PBMC) with antigen, and establishing the numbers of T cells producing the respective lead cytokine, IFN-γ and IL-2 for Th1 cells, IL-4 and IL-5 for Th2, and IL-17 for Th-17 cells, respectively. Traditionally, these cytokines are measured within 6 h in flow cytometry. We show here that 6 h of stimulation is sufficient to detect peptide-induced production of IFN-γ, but 24 h are required to reveal the full frequency of protein antigen-specific Th1 cells. Also the detection of IL-2 producing Th1 cells requires 24 h stimulation cultures. Measurements of IL-4 producing Th2 cells requires 48-h cultures and 96 h are required for frequency measurements of IL-5 and IL-17 secreting T cells. Therefore, accounting for the differential secretion kinetics of these cytokines is critical for the accurate determination of the frequencies and ratios of antigen-specific Th1, Th2, and Th17 cells. View Full-TextKeywords:
cytokine kinetics; ELISPOT; CD4 cells; T cells; immune monitoring; multiplexing
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MDPI and ACS Style
Duechting, A.; Przybyla, A.; Kuerten, S.; Lehmann, P.V. Delayed Activation Kinetics of Th2- and Th17 Cells Compared to Th1 Cells. Cells 2017, 6, 29.
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