Fig 2: The lung TNFR2+ cDC2 population maintains lung tolerance and prevents lung inflammation at steady state.a Numbers of lung CD4+ T cells at steady state in C57BL/6 J (n = 7), Batf3-/- (n = 6), CCR2-/- (n = 4), and IRF4fl/flCD11ccre (n = 7) mice. Data were compiled from two independent experiments. b Numbers of lung CD4+ T cells in WT (n = 12), IRF4fl/flCD11ccre (n = 10), RelBfl/flCD11ccre (n = 7), and TNFR2fl/flCD11ccre (n = 5) mice at steady state. Data were representative of two independent experiments. c Image of mediastinal lymph nodes (mLNs; top) and spleens (bottom) in of 7- to 8-week-old knockout strains at steady state; n = 3 mice/group. Data are representative of two independent experiments. d Numbers of CD45+ cells in the lungs (left), mLNs (center), and spleens (right) in the indicated strains at steady state; n = 3 mice/group. Data were representative of two independent experiments. e Numbers of CD4+ T cells in IRF4fl/flCD11ccre mice 14 days post cell adoptive transfer. TNFR2+ or TNFR2- cDC2 were sorted from WT mice lung and intranasally (i.n.) transferred into IRF4fl/flCD11ccre recipient mice; n = 4–6 mice/group. Data are representative of two independent experiments. f Numbers of lung Foxp3+CD4+ T-reg cells at steady state in C57BL/6 J, Batf3-/-, CCR2-/-, and IRF4fl/flCD11ccre mice; n = 4–5 mice/group. Data were representative of two independent experiments. Graphs represent the mean with error bars indication s.e.m. P values determined by one-way ANOVA Tukey’s multiple comparison test. *P < 0.05, **P < 0.001, and ***P < 0.0001.

Fig 3: Lung TNFR2+ cDC2 in healthy and lung diseases patients.a Flow cytometry analysis of pulmonary cDC2 subpopulation in healthy human lungs. Data are representative of eight independent experiments. b Flow cytometry analysis of TNFR2+cDC2 in donor, emphysema, and chronic-rejected human lungs. Data are representative of three independent experiments. c Frequency of TNFR2+ cDC2 in diseased human lungs. Data were compiled from multiple independent experiments. Graphs represent the mean with error bars indication s.e.m.

Fig 4: A pathogenic role of the TNFa-TNFRI axis during T cell-mediated autoinflammation. a Staining of MEFs co-cultured with T cells. Scale bars: 100 µm. Magnified images are shown on the right. b–d Percentage of surviving cells in a (b), Sharpin-/- MEFs co-cultured with the indicated T cells (c), and the indicated CRISPR/Cas9-targeted MEFs (d) n = 4 biologically independent experiments. e Treatment with a TNFa-neutralizing antibody or solubilized TNFRII. f Treatment with z-VAD-fmk and/or Nec1. The iCelligence system was used to measure cell viability. g, h Survival after 10 h of the treatment described in f (g) or after siRNA knockdown (h). n = 4 independent experiments. i Expression of the indicated receptors on the T cell surface after TCR stimulation. j Disease progression among the indicated strains. M, Male; F, Female. k Immunofluorescence staining of skin sections. Scale bars: 20 µm. l Ratio of CD3eintTCRß+:CD3ehiTCR?d+ cells. n = 6 biologically independent animals. m Number of TCRß+ T cells in the skin. n = 4 biologically independent animals. n Percentage of TNFa- and/or IFN?-expressing TCRß+ T cells in the skin. n = 4 biologically independent animals. o Dot plots representing TRM cells. p, q MFI or percentage of cells expressing the indicated surface proteins. n = 8 biologically independent animals. Small horizontal lines indicate the mean (±s.e.m.). *p < 0.05, **p < 0.01, ***p < 0.001. Kruskal–Wallis test with Bonferroni correction (a value = 0.05) was used for b–d, g, and h, while two-tailed Mann–Whitney U-test was used for l–n, p, and q. Data are representative of at least two independent experiments (a, f, i, k, o), or pooled from three to six (b–e, g, h, l–n, p, q) independent experiments. Source data are provided in a Source Data file

Fig 5: Decreased cell surface expression of TNFR1 in the TRAPS mutant cells. The expression of TNFR1 and TNFR2 were determined in primary murine bone marrow-derived macrophages and peritoneal macrophages. (A) mRNA expression levels of Tnfrsf1a and Tnfrsf1b in bone marrow-derived macrophages. (B) Immunoblot analysis of TNFR1 and TNFR2 expression in the bone marrow-derived macrophages. Two different antibodies against TNFR1 were used: Ab#1 (13377, Cell Signaling Technology) recognizes aa 29–43 (extracellular domain), and Ab#2 (AF-425-PB, R&D Systems) recognizes the C-terminal intracellular region. (C–E) Flow cytometric analysis of TNFR1 and TNFR2 expression. Peritoneal exudate cells were collected from the indicated mice 3 days after the intraperitoneal administration of thioglycolate. (C) Representative histograms of the cell surface expression of TNFR1 and TNFR2. The expression levels of TNFR1 (D) and TNFR2 (E) on the surface of CD11b-positive cells were determined by flow cytometry. The cells within the horizontal line on each histogram were recognized as TNFR1- or TNFR2-positive cells. The proportions of the positive cells were calculated relative to those of the WT in each strain. The following antibodies were used: TNFR1 (113005, BioLegend) and TNFR2 (113405, BioLegend). Both antibodies recognize the extracellular domains. (F, G) ELISA for soluble TNFR1 (sTNFR1) in the culture supernatant of bone marrow-derived macrophages. Culture supernatant was collected before (F) and 24 h after (G) LPS stimulation. Concentrations of sTNFR1 in the culture supernatant were measured by ELISA, and the levels were calculated relative to that of the WT in each strain. TRAPS, TNF receptor-associated periodic syndrome; LPS, lipopolysaccharide; ELISA, enzyme-linked immunosorbent assay; WT, wild-type; Het, heterozygote; Hom, homozygote.