Fig 2: Dectin-1 does not instruct IFN-ß-mediated non-pathogenic TH17 polarization via IL-1ß, IL-6, IL-23, IL-12 or IL-27.a,b, ELISA for quantification of IL-1ß, IL-6, IL-23, IL-12 and IL-27 in the supernatant of unstimulated DCs or after stimulation of DCs with curdlan or C. albicans, in the presence of blocking IFN-a/ßR antibodies (a,b) or rhIFN-ß (b) at 28 h (a, n = 5; b, IFN-a/ßR antibody n = 5, rhIFN-ß n = 3). Data in a,b represent mean ± s.d. of independent donors. **P < 0.01, *P < 0.05 (paired, two-tailed Student’s t-test), calculated between untreated and treated samples that were likewise stimulated. c–e, Flow cytometry analyses of TH polarization by staining for intracellular IL-17 and IFN-? expression (FI) in restimulated T cells, outgrown in vitro by co-culture of memory CD4+ T cells with iDC or DCs primed for 48 h with curdlan or C. albicans, in the presence of neutralizing IL-27 antibodies or rhIL-27 (both during DC-T co-culture), at day 11–14 (IL-27 antibody n = 7, rhIL-27 n = 4). In c, representative dot plots for independent donors are shown, with percentage positive cells indicated in each quadrant. In d, the percentage IL-17+ cells per total amount of T cells are shown and in e, the percentage IFN-?- and percentage IFN-?+ cells per IL-17+ T cells are shown. Data in d,e represent mean ± s.d. of independent donors.Source data

Fig 3: CAF-induced CD39 expression can be inhibited by neutralizing IFN-? and TNF-a or by inhibition of TGF-ß signaling a) Expression of CD39, LAG-3, PD-1 and Tim3 on CD8 + T cells activated either alone (PBMC-only) or in co-culture with CAF (+CAF) in the presence of neutralizing antibody together IL-6 or IL-27. b) Expression of CD39, CD49b, Tim3, PD-1 LAG3 and CD103 on activated CD8+ T cells either cultured alone (media) or in co-culture with CAF (+CAF) and neutralizing antibodies to IFN-? and TNF-a, anti-PD-L1, anti-PD-1, SB431542, SB431542+ anti-PD-1 or isotype control antibody as indicated. Error bars show standard deviation of triplicate wells. Significance shown for each parameter when compared to +CAF. Results for SB431542 are representative of those from three repeat experiments using different PBMCs. For PD-1, NA where PD-1 blockage used with PD-1 surface levels. One way ANOVA was used for all statistical analysis with Tukey’s multiple comparisons posttest (ns = not significant,* P = <0.05, ** P = <0.01, *** P = <0.001, ****P = <0.0001)

Fig 4: Cytokines produced by activated T cells promote production of IL-6 and IL-27 by CAF. IL-6 production a) by five independently generated NSCLC CAF lines co-cultured with either PBMCs only (+PBMC), with PBMCs +anti-CD3/anti-CD28 antibodies (+PBMC+STIM) or with PBMC+anti-CD3/anti-CD28 antibodies and neutralizing cytokines to IFN-? and TNF-a. b) IL-6 production by CAF cultured in the presence of conditioned media from anti-CD3 stimulated tumor infiltrating T cells (TCM) pre-incubated with neutralizing antibodies to either IFN-?, TNF-a or both IFN-?/TNF-a. c) IL-6 production by CAF in response to rIFN-?, rTNF-a (each at 25 ng/ml) or a combination of rIFN-? and rTNF-a. d) IL-27 production in cultures of PBMC stimulated with anti-CD3/anti-CD28 alone or when in co-culture with CAF. E) IL-27 production by seven independently generated NSCLC CAF lines stimulated with rIFN-? or rTNF-a (both used at 25 ng/ml) or a combination of IFN-? and TNF-a. Results are pooled from three experiments. One-way ANOVA was used for all statistical analysis with Tukey’s multiple comparisons posttest (* P = <0.05, ** P = <0.01, *** P = <0.001, ****P = <0.0001)

Fig 5: HBV-specific as well as global IL-27 expressing TFH cells were augmented in PD-1+ fraction of cells rather than PD-1-. (A) HBV-specific as well as global cytokine producing TFH cells were assessed in PD-1+ and PD-1- TFH cell population in CHB and HC-vacc (n=14 subjects in each group). (B) To identify the functional status of PD-1+ TFH cells, expression of CD69, an activation marker, was analyzed on both PD-1+ and PD-1- cells after HBs, HBc and PMA/ionomycin stimulation. Statistics were calculated using 2-tailed non-parametric, Mann-Whitney U test. Bars graphs indicate median with range. * indicates p < 0.05 and ***p < 0.001.