CD138 expression is a molecular signature but not a developmental requirement for RORγt+ NKT17 cells
Shunqun Luo, … , Pyong Woo Park, Jung-Hyun Park
Shunqun Luo, … , Pyong Woo Park, Jung-Hyun Park
Published September 22, 2021
Citation Information: JCI Insight. 2021;6(18):e148038. https://doi.org/10.1172/jci.insight.148038.
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Research Article Immunology

CD138 expression is a molecular signature but not a developmental requirement for RORγt+ NKT17 cells

Abstract

Invariant NKT (iNKT) cells are potent immunomodulatory cells that acquire effector function during their development in the thymus. IL-17–producing iNKT cells are commonly referred to as NKT17 cells, and they are unique among iNKT cells to express the heparan sulfate proteoglycan CD138 and the transcription factor RORγt. Whether and how CD138 and RORγt contribute to NKT17 cell differentiation, and whether there is an interplay between RORγt and CD138 expression to control iNKT lineage fate, remain mostly unknown. Here, we showed that CD138 expression was only associated with and not required for the differentiation and IL-17 production of NKT17 cells. Consequently, CD138-deficient mice still generated robust numbers of IL-17–producing RORγt+ NKT17 cells. Moreover, forced expression of RORγt significantly promoted the generation of thymic NKT17 cells, but did not induce CD138 expression on non-NKT17 cells. These results indicated that NKT17 cell generation and IL-17 production were driven by RORγt, employing mechanisms that were independent of CD138. Therefore, our study effectively dissociated CD138 expression from the RORγt-driven molecular pathway of NKT17 cell differentiation.

Authors

Shunqun Luo, Juntae Kwon, Assiatu Crossman, Pyong Woo Park, Jung-Hyun Park

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Figure 3

Functional and phenotypical characterization of Sdc1–/– iNKT cells.

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Functional and phenotypical characterization of Sdc1–/– iNKT cells. (A) ...
(A) Identification of thymic NKT17 cells based on CD122 and CD4 expression. CD138+ and CD138– iNKT cells were assessed for surface CD122 and CD4 expression. Data are representative of 5 independent experiments with a total of 7 BALB/c mice. (B) iNKT subset classification based on CD122 and CD4 expression. Thymic iNKT cells of Sdc1–/– and WT littermate BALB/c thymocytes were assessed for CD122 and CD4 expression, visualizing the 3 subsets of NKT1 (CD122+), NKT2 (CD122–CD4+), and NKT17 (CD122– CD4–) cells (left). CD138 expression was assessed in the indicated iNKT subsets of Sdc1–/– and WT littermate BALB/c thymocytes. Data are representative of 2 independent experiments. (C) iNKT subsets were identified in fixed and permeabilized thymocytes of Sdc1–/– and WT littermate BALB/c mice based on CD122 and CD4 expression (left). Intracellular perforin and granzyme A expression were then assessed in each of the indicated iNKT subsets. Contour plots and histograms are representative of 3 independent experiments. (D) Surface CD69 and CD25 expression was assessed on NKT17 cells upon overnight in vitro stimulation of Sdc1–/– and WT littermate BALB/c thymocytes with the indicated amounts of α-GalCer. Contour plots are representative and bar graph shows the summary of 3 independent experiments. All data are presented as mean ± SEM. P values were determined by unpaired 2-tailed Student’s t test. NS, not significant.