Peripheral T Cell Populations are Differentially Affected in Familial Mediterranean Fever, Chronic Granulomatous Disease, and Gout

Abstract

Both innate errors of immunity, such as familial Mediterranean fever (FMF) and chronic granulomatous disease (CGD), and the common inflammatory disease gout are characterized by episodes of sterile inflammatory attacks in the absence of an infection. While these disorders encompass distinct pathologies due to differentially affected metabolic pathways and inflammasome activation mechanisms, their common features are the excessive production of interleukin (IL)-1ß and innate immune cell hyperreactivity. On the other hand, the role of T cells and innate-like lymphocytes such as gamma delta (γδ) T cells in these pathologies is ill-defined. In order to widen our understanding of T cell involvement in CGD, FMF and gout pathology, we developed multicolour immunophenotyping panels for flow cytometry to characterize γδ T cells as well as CD4 and CD8 T cell populations in terms of their cytokine production, activation status, memory or naive phenotypes, exhaustion status, homing receptor expression, and cytotoxic activity. Our study is the first deep immunophenotyping analysis of T cell populations in CGD, FMF, and gout patients. We found that CGD affects the frequencies and activation status of T cells, while gout impairs the cytokine production capacity of Vδ2 T cells. FMF was characterized by decreased percentages of regulatory T cells in circulation and attenuated IFN-γ production capacity by Vδ2 T cells. Autoinflammatory syndromes and congenital defects of phagocyte differentially affect T cell compartments. Future studies are warranted to assess whether these phenotypical changes are relevant for disease pathology.

Keywords: Familial Mediterranean fever; T cells; chronic granulomatous disease; flow cytometry; gamma delta T cells; gout; immunophenotyping; inborn errors of immunity.

Fig. 1

The distribution of circulating lymphocyte populations is significantly affected in CGD patients. a–h Percentages of different cell populations: CD3⁺ TCRVδ1⁺ cells (a), CD3⁺ TCRVδ2⁺ cells (b), CD3⁺ CD4⁺ cells (c), CD3⁺ CD8⁺ cells (d), CD3⁺ CD56⁺ NKT-like cells (e), CD3⁺ CD4⁺ CD25^hi CD127⁻ regulatory T cells (f), CD3⁻ CD19⁺ B cells (g), and CD3⁻ CD56⁺ NK cells (h) in freshly isolated PMBCs from healthy controls and patients determined by flow cytometry. CGD chronic granulomatous disease (n = 5), FMF familial Mediterranean fever (n = 7), gout (n = 6). Gray circle: FMF patient with Behcet disease. Non-parametric Mann–Whitney test was applied. *p ˂ 0.05, **p ˂ 0.01

Fig. 2

CGD results in increased expression of activation markers on Vδ2 and CD8 T cells. a–f Percentages of Vδ2 (a, d), CD4 (b, e), and CD8 T cells (c, f) expressing CD38 (a–c) and CD69 (d–f) assessed by flow cytometry on freshly isolated PMBCs from healthy controls and patients. CGD chronic granulomatous disease (n = 5), FMF familial Mediterranean fever (n = 7), gout (n = 6). Gray circle: FMF patient with Behcet disease. Non-parametric Mann–Whitney test was applied for statistical analysis. *p ˂ 0.05, **p ˂ 0.01

Fig. 3

CGD and FMF affect PD-1 and CD95 expression on T cells. a–i Percentages of Vδ2 (a, d, g), CD4 (b, e, h), and CD8 T cells (c, f, i) expressing CD95 (a–c), PD-1 (d–f), and CTLA-4 (g–i) assessed by flow cytometry in freshly isolated PMBCs from healthy controls and patients. CGD chronic granulomatous disease (n = 5), FMF familial Mediterranean fever (n = 7), gout (n = 6). Gray circle: FMF patient with Behcet disease. Non-parametric Mann–Whitney test was applied for statistical analysis. *p ˂ 0.05

Fig. 4

FMF and gout patients show reduced numbers of cytokine-producing Vδ2 T cells. a–f Percentages of IFN-γ- (a–c) and TNF-α- (d–f) producing Vδ2 (a, d), CD4 (b, e), and CD8 T cells (c, f) assessed by flow cytometry in freshly isolated PMBCs from healthy controls and patients. CGD cChronic granulomatous disease (n = 5), FMF familial Mediterranean fever (n = 7), gout (n = 6). Gray circle: FMF patient with Behcet disease. Non-parametric Mann–Whitney test was applied for statistical analysis. *p ˂ 0.05, **p ˂ 0.01

Fig. 5

Homing receptors expression on peripheral T cells is not affected in CGD, FMF and gout. a–l Percentages of Vδ2 (a, d, g, j), CD4 (b, e, h, k), and CD8 T cells (c, f, i, l) expressing CCR2 (a, b, c), CCR5 (d, e, f), CCR7 (g, h, i), and CCR8 (j, k, l) assessed by flow cytometry in freshly isolated PMBCs from healthy controls and patients. CGD chronic granulomatous disease (n = 5), FMF familial Mediterranean fever (n = 7), gout (n = 6). Gray circle: FMF patient with Behcet disease. Non-parametric Mann–Whitney test was applied. *p ˂ 0.05

Fig. 6

T cells exhibit no change in expression of cytotoxicity markers in FMF, CGD, and gout. a–g Expression levels of CD54 (a–c), and CD56 (d, e) and NKG2D (f, g) on peripheral Vδ2 (a, d, f), CD4 (b), and CD8 T (c, e, g) assessed by flow cytometry on freshly isolated PMBCs from healthy controls and patients. Dashed lines in y-axis show the MFI of fluorescence minus one (FMO) control. CGD chronic granulomatous disease (n = 5), FMF familial Mediterranean fever (n = 7), gout (n = 6). Gray circle: FMF patient with Behcet disease. Non-parametric Mann–Whitney test was applied for statistical analysis. *p ˂ 0.05, **p ˂ 0.01, ***p < 0.001