ICOS regulates IL-10 production in group 2 innate lymphoid cells via cholesterol and cortisol biosynthesis
Yoshihiro Sakano, Kei Sakano, Benjamin P. Hurrell, Mohammad H. Kazemi, Xin Li, Stephen Shen, Omid Akbari
Yoshihiro Sakano, Kei Sakano, Benjamin P. Hurrell, Mohammad H. Kazemi, Xin Li, Stephen Shen, Omid Akbari
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Research Article Immunology Pulmonology

ICOS regulates IL-10 production in group 2 innate lymphoid cells via cholesterol and cortisol biosynthesis

Abstract

Group 2 innate lymphoid cells (ILC2s) play a crucial role in inducing type 2 inflammation in the lungs in response to allergens. Our study investigated the regulatory mechanism of IL-10 production by ILC2s and its impact on airway hyperreactivity (AHR), focusing on the role of ICOS. We found that inhibiting ICOS in pulmonary ILC2s significantly enhanced IL-10 production. The absence of ICOS reprogrammed ILC2 steroid metabolism, leading to increased cholesterol and cortisol biosynthesis and subsequent glucocorticoid receptor (GR) activation. This reprogramming regulated MAF and NFIL3 activation, promoting IL-10 production. Notably, in vivo GR inhibition or ILC2-specific GR deficiency exacerbated AHR development in multiple mouse models. We extended these findings to human ILC2s, demonstrating concordant results between murine models and human cells. Our results indicate that ICOS negatively regulates IL-10 production in ILC2s by controlling cholesterol and cortisol biosynthesis. This mechanism provides new insights into the complex interplay between ILC2s, ICOS, and glucocorticoid signaling in the context of allergic airway inflammation.

Authors

Yoshihiro Sakano, Kei Sakano, Benjamin P. Hurrell, Mohammad H. Kazemi, Xin Li, Stephen Shen, Omid Akbari

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Figure 6

Cortisol is a key regulator of ICOS-mediated IL-10 production in ILC2s.

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Cortisol is a key regulator of ICOS-mediated IL-10 production in ILC2s. ...
(A and B) Total RNA from WT and ICOS-KO mice was extracted to perform a bulk transcriptomic analysis. (A) Volcano plots represent differentially expressed genes involved in steroid biosynthesis. (B) Biosynthetic pathway of cortisol from cholesterol showing intermediate metabolites and key enzymes involved in each step. Changes in enzyme expression are indicated as log2(fold change) values in colored boxes. (C) The bar graph represents the normalized counts of Cyp11a1. n = 3. (D) Representative histogram of protein expression of CYP11A1. Corresponding quantitation is presented as MFI. n = 4. (E) WT and ICOS-KO aILC2s were isolated and cultured with or without U18666A or statin. (F) Levels of cortisol in the culture supernatant were measured by ELISA. n = 3. (G) Representative histogram of protein expression of GR. Corresponding quantitation is presented as MFI. n = 4. (H–K) ICOS-KO aILC2s were cultured with or without cortisol. (I) Levels of IL-10 in the culture supernatant were measured. n = 4. (J and K) Representative plots of MAF (J) and NFIL3 (K) expression levels in each group are shown. Corresponding quantitation is presented as MFI. n = 4. (L–O) Sorted aILC2s from NMUR1cre (ILC2WT mice) and NMUR1creGRfl/fl mice (ILC2ΔGR mice) were cultured with isotype control or anti-ICOS antibody. (M) Bar graph representing the frequency of IL-10+ ILC2s in each group. n = 4. (N and O) Representative plots of MAF (N) and NFIL3 (O) expression levels in each group are shown. Corresponding quantitation is presented as MFI. n = 4. Data are presented as mean ± SEM and are representative of 4 independent experiments. Two-tailed Student’s t test was employed for statistical analysis; *P < 0.05, **P < 0.01, and ***P < 0.001, and ****P < 0.0001. Schematic images were created in Adobe Illustrator. FMO, fluorescence minus one.