Journal: Nature Communications

Article Title: T cell-intrinsic IL-1R signaling licenses effector cytokine production by memory CD4 T cells

doi: 10.1038/s41467-018-05489-7

Figure Lengend Snippet: Effector CD4 T Cells constitutively express IL-1R and require IL-1R signaling for cytokine production. a , b Splenic CD4 T cells were gated on naive (T naive ) and memory (T em ) T cell markers and stained for a IL-6Ra, IL-4Ra and IL-12Rb1 b IL-1R1, IL-18Rα and IL-33R (ST2). c Relative expression of Il1r1 transcripts in T naive and T em cells. Data is normalized to 18s. d - f T em cells were stimulated with splenic DCs and soluble αCD3 (30 ng ml −1 ) in the presence or absence of IL-1R antagonist (IL-1Ra) for 48 h d IL-17A, IL-17F, IL-22, e IL-13, IL-4, IL-5, and f IFNγ were measured. g CD4 T cells from SI-LP were stained for IL-1R1. h CD4 T cells from SI-LP were stimulated using CD11C + DCs and soluble αCD3 (30 ng ml −1 ) in the presence or absence of IL-1Ra for 48 h followed by cytokine ELISA. i IL-17A, IL-13 and IFNγ levels in supernatants obtained from T em cell co-cultures with WT or Il1r −/− CD11c + splenic DCs in the presence of αCD3 (30 ng ml −1 ) after 48 h of stimulation. Control in receptor staining refers to fluorescence minus one control. IL-17A, IL-13 and IFNγ in the culture supernatants were measured using paired-antibody ELISA. Error bars indicate SEM; paired t test, c – f , h , i 4–8 mice were pooled per experiment

Article Snippet: Mouse CD4 BV421 Biolegend 100443 (1:400), Mouse CD4 FITC Biolegend 100405 (1:400), CD44 AF700 Biolegend 103025 (1:400), CD44 pacific blue Biolegend 103020 (1:400), CD62L APC Tonbo Biosciences 20-0621-U100 (1:400), IL-6r Biotin Biolegend 115803 (1:100), IL-12rb1 PE RnD systems FAB1998P-025 (10ul/test), IL-4Ra PE Biolegend 144803 (1:100), IL-1R1 Biotin Biolegend 113503 (1:100), IL-18Ra APC Biolegend 132903 (5ul/test), IL-17A FITC Biolegend 506908 (1:300), IL-22 PE Biolegend 516404 (5ul/test), IFNg PE Biolegend 505807 (1:200), ICOS APC Biolegend 313509 (1:400), CD69 PE BD Biosciences 553237 (1:400), CD25 Biotin BD Biosiences (1:400), Zombie Yellow live dead stain Biolegend 423103 (1:500), Streptavidin BV421 Biolegend 405241 (1:400), CFSE Biolegend 423801 (1:1000), IL-17A purified Biolegend 505807 (1:1000), antiIL-17A biotin Biolegend 507002 (1:1000), Anti IL-22 purified Biolegend 516401 (1:250), Anti IL-22 biotin Biolegend 516407 (1:200), Anti IFNg purified Biolegend 505702 (1:1000), Anti IFNg biotin Biolegend 505804 (1:1000), Anti IL-17F Capture antibody eBioscience 14-7473-68 (1:200), Anti IL-17F Detection antibody eBioscience 13-7474-68 (1:250), Anti IL-13 purified eBioscience 14-7133-85 (1:125), Anti IL-13 biotin eBioscience 13-7135-85 (1:2000), Anti IL-4 purified Biolegend 504102 (1:200), Anti IL-4 biotin Biolegend 504202 (1:2000), Anti IL-5 purified Biolegend 504301 (1:500), Anti IL-5 biotin Biolegend 504402 (1:500)

Techniques: Staining, Expressing, Enzyme-linked Immunosorbent Assay, Fluorescence

Journal: Nature Communications

Article Title: T cell-intrinsic IL-1R signaling licenses effector cytokine production by memory CD4 T cells

doi: 10.1038/s41467-018-05489-7

Figure Lengend Snippet: IL-1β licenses IL-17A, IFNγ and IL-13 production by circulating effector CD4 T cells. a CD44 hi CD62L lo T em cells were stimulated with immobilized αCD3 (0.5 μg ml −1 ) and αCD28 (0.5 μg ml −1 ) in the presence of indicated concentrations of recombinant IL-1α (left) and IL-1β (right). b , c T em cells were co-cultured with splenic DCs and soluble αCD3 (30 ng ml −1 ) in the presence or absence of neutralizing antibodies against IL-1α or IL-1β (10 μg ml −1 ). d T em cells from WT mice were stimulated with soluble αCD3 (30 ng ml −1 ) in the presence of splenic DCs from indicated genotypes. e T em cells were stimulated with αCD3 (30 ng ml −1 ) using either splenic DCs or naive B cells as APCs in the presence or absence of recombinant IL-1β (10 ng ml −1 ). Cytokine concentrations in the culture supernatants were determined using paired-antibody ELISA. Error bars indicate SEM; paired t test

Article Snippet: Mouse CD4 BV421 Biolegend 100443 (1:400), Mouse CD4 FITC Biolegend 100405 (1:400), CD44 AF700 Biolegend 103025 (1:400), CD44 pacific blue Biolegend 103020 (1:400), CD62L APC Tonbo Biosciences 20-0621-U100 (1:400), IL-6r Biotin Biolegend 115803 (1:100), IL-12rb1 PE RnD systems FAB1998P-025 (10ul/test), IL-4Ra PE Biolegend 144803 (1:100), IL-1R1 Biotin Biolegend 113503 (1:100), IL-18Ra APC Biolegend 132903 (5ul/test), IL-17A FITC Biolegend 506908 (1:300), IL-22 PE Biolegend 516404 (5ul/test), IFNg PE Biolegend 505807 (1:200), ICOS APC Biolegend 313509 (1:400), CD69 PE BD Biosciences 553237 (1:400), CD25 Biotin BD Biosiences (1:400), Zombie Yellow live dead stain Biolegend 423103 (1:500), Streptavidin BV421 Biolegend 405241 (1:400), CFSE Biolegend 423801 (1:1000), IL-17A purified Biolegend 505807 (1:1000), antiIL-17A biotin Biolegend 507002 (1:1000), Anti IL-22 purified Biolegend 516401 (1:250), Anti IL-22 biotin Biolegend 516407 (1:200), Anti IFNg purified Biolegend 505702 (1:1000), Anti IFNg biotin Biolegend 505804 (1:1000), Anti IL-17F Capture antibody eBioscience 14-7473-68 (1:200), Anti IL-17F Detection antibody eBioscience 13-7474-68 (1:250), Anti IL-13 purified eBioscience 14-7133-85 (1:125), Anti IL-13 biotin eBioscience 13-7135-85 (1:2000), Anti IL-4 purified Biolegend 504102 (1:200), Anti IL-4 biotin Biolegend 504202 (1:2000), Anti IL-5 purified Biolegend 504301 (1:500), Anti IL-5 biotin Biolegend 504402 (1:500)

Techniques: Recombinant, Cell Culture, Enzyme-linked Immunosorbent Assay