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. 2023 Jun;51(6):3000605231180039.
doi: 10.1177/03000605231180039.

Effects of interleukin-23 on the activation of mucosal-associated invariant T cells from oral lichen planus

Fang Wang  1   2 , Xiu-Li Zhang  1 , Jing-Ya Yang  1   2 , Gang Zhou  1   2
Affiliations

Effects of interleukin-23 on the activation of mucosal-associated invariant T cells from oral lichen planus

Fang Wang et al. J Int Med Res. 2023 Jun.

Abstract

Objective: Oral lichen planus (OLP) is a T cell-mediated inflammatory condition in the oral cavity. Mucosal-associated invariant T (MAIT) cells are gaining more relevance in immune diseases because they can be activated by cytokines without T cell receptor stimulation. Herein, we tested the effect of interleukin-23 (IL-23) on the activation status of OLP MAIT cells.

Methods: Peripheral blood mononuclear cells (PBMCs) isolated from OLP patients were stimulated by IL-23 in the absence or presence of phorbol myristate acetate (PMA) and ionomycin. The activation status of MAIT cells was analyzed by flow cytometry after staining for CD3, CD4, CD8, CD161, TCR Vα7.2, and CD69.

Results: The fraction of MAIT cells in OLP peripheral blood was approximately 0.38% to 3.97%, and CD8+ subpopulations overwhelmed CD4+ cells. The mean percentages of OLP MAIT cells in PBMCs and CD8+MAIT cells in MAIT cells were approximately 40%. PMA and ionomycin significantly increased CD69 expression on OLP T cells, MAIT cells, and CD8+MAIT cells. Cells with enhanced activation had different responsiveness to exogenous IL-23, showing increased CD69 expression on OLP T cells, decreased CD69 on OLP CD8+MAIT cells, and no significant change on OLP MAIT cells.

Conclusions: IL-23 showed different effects on the activation status of OLP MAIT cells and CD8+MAIT cells.

Keywords: CD69; Mucosal-associated invariant T cell; interleukin-23; ionomycin; oral lichen planus; phorbol myristate acetate.

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Conflict of interest statement

The authors declare that there is no conflict of interest.

Figures

Figure 1.
Figure 1.
Effect of IL-23 on the proportion of MAIT cells among PBMCs from OLP patients in the absence or presence of PMA and ionomycin. Briefly, 1 × 106 PBMCs were cultured per well in 24-well plates and stimulated with or without 10 ng/mL IL-23 for 72 hours in the absence or presence of 50 ng/mL PMA and 1 μg/mL ionomycin (IM). (a–d) The gating strategy for T cells, MAIT cells, and MAIT subsets are shown by representative images in different groups. (e–g) Histograms of the proportions of T cells, MAIT cells, and MAIT subsets. Normally distributed variables are presented as mean ± SD, while non-normally distributed data are presented as mean ± IQR and labeled by “#”. P < 0.05 was defined as statistically significant. MAIT, mucosal-associated invariant T; PBMCs, peripheral blood mononuclear cells; OLP, oral lichen planus; PMA, phorbol myristate acetate; SD, standard deviation; IQR, interquartile range.
Figure 2.
Figure 2.
The effect of IL-23 on the expression of CD69 in MAIT cells from OLP patients in the absence or presence of PMA and ionomycin. (a) The gating strategy for CD69 staining in T cells, MAIT cells, and CD8+MAIT cells is shown by representative images in different groups and (b–d) Histograms of CD69 expression in T cells, MAIT cells, and CD8+MAIT cells. Normally distributed variables are presented as mean ± SD, while non-normally distributed data are presented as mean ± IQR and labeled by “#”. P < 0.05 was defined as statistically significant. MAIT, mucosal-associated invariant T; OLP, oral lichen planus; PMA, phorbol myristate acetate; FMO, fluorescence minus one; SD, standard deviation; IQR, interquartile range.
Figure 3.
Figure 3.
The effect of IL-23 on the production of IFN-γ and IL-17 by MAIT cells and CD8+MAIT cells from OLP patients in the absence or presence of PMA and ionomycin. (a) The gating strategy for IFN-γ and IL-17 staining in MAIT cells and CD8+MAIT cells is shown by representative images in different groups and (b–e) Histograms of IFN-γ and IL-17 expression in MAIT cells and CD8+MAIT cells. Normally distributed variables are presented as mean ± SD. P < 0.05 was defined as statistically significant. MAIT, mucosal-associated invariant T; OLP, oral lichen planus; PMA, phorbol myristate acetate; FMO, fluorescence minus one; SD, standard deviation.
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