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. Author manuscript; available in PMC: 2017 Oct 1.
Published in final edited form as: J Immunol. 2017 Feb 17;198(7):2699–2711. doi: 10.4049/jimmunol.1601202

Figure 6.

Figure 6

ITK-deficient CTLs exhibit reduced TCR-triggered degranulation. Degranulation measured in a flow-based LAMP1 cycling assay in WT (black) or Itk−/− (grey) CTLs in response to (A) plate-bound anti-CD3, and 1μM OVA257-264-pulsed (B) LPS-activated B cell or (C) EL4 targets. Each line represents data from paired mice from an independent LAMP1 cycling experiment, where CD107a-PE positive cells in the CD8+ gate were determined to be positive for degranulation. *P<0.05, **P<0.01 calculated by paired sample t-tests. (D) Degranulation in response to 5μg/mL plate-bound anti-CD3 in day 6 WT OT-I CTLs (black solid bar), WT OT-I CTLs pre-treated with the ITK inhibitor, 10n, at 0.03 μM (horizontal striped bar), 0.1 μM (vertical striped bar), 0.2 μM (black and grey diagonal bar) or 0.3 μM (black and white inverse diagonal bar), or Itk−/− OT-I CTLs (grey solid bar). Graph representative of two independent experiments. (E) Degranulation in allo-reactive human CD8+ T cells from two healthy donors treated with increasing concentrations of 10n in response to 5μg/mL of plate-bound OKT3. Graph represents data using the same donors as in Figure 3E and is representative of one of two independent experiments.