IFN-γ–producing self-reactive CD4+ T cells induce autoimmune adrenalitis in a mouse model of Addison’s disease
Arina Andreyeva, Juraj Michalik, Veronika Niederlova, Veronika Cimermanova, Ales Drobek, Radislav Sedlacek, Jan Prochazka, Juraj Labaj, Olha Fedosieieva, Waldemar Kanczkowski, Peter Draber, André Sulen, Ondrej Stepanek, Aleš Neuwirth
Arina Andreyeva, Juraj Michalik, Veronika Niederlova, Veronika Cimermanova, Ales Drobek, Radislav Sedlacek, Jan Prochazka, Juraj Labaj, Olha Fedosieieva, Waldemar Kanczkowski, Peter Draber, André Sulen, Ondrej Stepanek, Aleš Neuwirth
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Research Article Clinical Research Endocrinology

IFN-γ–producing self-reactive CD4+ T cells induce autoimmune adrenalitis in a mouse model of Addison’s disease

Abstract

Autoimmune Addison’s disease (AD) is a rare but life-threatening disorder caused by immune-mediated destruction of the adrenal cortex, and progress in therapy has been limited by insufficient mechanistic insight. Here, we establish a model of experimental autoimmune adrenalitis that recapitulates key features of AD and reveals sex-dependent differences in disease manifestation within the model. Immunization with peptides derived from the adrenal self-antigen CYP11A1 induces corticosterone insufficiency. We show that autoimmune adrenalitis is driven by IFN-γ produced by self-reactive CD4+ T cells, promoting granulomatous inflammation in the adrenal cortex. Together, these findings identify IFN-γ as a central effector of autoimmune adrenalitis and suggest that targeting the IFN-γ pathway may represent a potential therapeutic strategy for AD.

Authors

Arina Andreyeva, Juraj Michalik, Veronika Niederlova, Veronika Cimermanova, Ales Drobek, Radislav Sedlacek, Jan Prochazka, Juraj Labaj, Olha Fedosieieva, Waldemar Kanczkowski, Peter Draber, André Sulen, Ondrej Stepanek, Aleš Neuwirth

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Figure 2

Adrenal CD4+ T cells are clonally expanded and include CYP11A1-reactive clones.

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Adrenal CD4+ T cells are clonally expanded and include CYP11A1-reactive ...
(A) UMAP of CD4+ T cells from adrenal glands and spleens of pCYP11A1-immunized mice showing expanded T cell clones (≥2 cells sharing the same TCRαβ). (B) Distribution of expanded CD4+ T cell clones between adrenal glands and spleens. (C) Distribution of expanded CD4+ T cell clones across defined T cell subsets. Clone size was calculated separately for each subset. (A–C) Color indicates clone size (number of cells sharing the same TCRαβ). (D) Composition of the 20 largest expanded CD4+ T cell clones. Clones are arranged by effector phenotype diversity, from polarized (left) to multidirectional (right); cell numbers are indicated on the x axis. Colors indicate effector phenotype; dominant phenotypes (≥50%) are shown above. (E) UMAP highlighting 4 expanded CD4+ T cell clones (ADR1–ADR4) among effector populations. Colors indicate clone identity. (F) Representative histograms of NFAT-GFP reporter activation in A5 hybridomas expressing ADR1–ADR4 TCRs stimulated with serial dilutions of CYP11A1150-164 or CYP11A1233-247. B3K508 A5 hybridomas stimulated with 3K peptide served as a positive control, whereas peptide-free cultures and irrelevant peptides at the highest concentration served as negative controls. PMA/ionomycin served as a positive stimulation control. Colors indicate peptide concentration. Representative of 3 independent experiments. (G) Quantification of GFP+ cells corresponding to F. Data are mean ± SEM from 3 independent experiments. Colors indicate individual T cell clones. (H) Experimental design for testing antigen specificity of 2 retrogenic CD4+ T cell clones. CD69 and CD25 expression was measured after stimulation with CYP11A1150-164 or CYP11A1233-247; peptide-free cultures served as controls. Representative of 3 independent experiments. Created with BioRender.com.