ril 17a  (R&D Systems)

Journal: Science Advances

Article Title: Pregnancy enhances antiviral immunity independent of type I IFN but dependent on IL-17–producing γδ + T cells in the nasal mucosa

doi: 10.1126/sciadv.ado7087

Figure Lengend Snippet: ( A ) Model of murine intratracheal IAV infection (50 PFU in 50 μl of PBS). Created with BioRender.com . ( B ) Kinetics of pulmonary viral titers by MDCK plaque assay following intratracheal IAV infection ( n = 7 to 9). ( C ) Graphical representation of upper airway infection (50 PFU IAV in 10 μl of PBS). Created with BioRender.com . ( D ) Kinetics of the viral load measured by MDCK plaque assay on nasal tissues from WT nonpregnant and pregnant mice ( n = 7 to 10). ( E ) Graphical representation of nasal tissue regions and fluorescence microscopy of infected olfactory epithelial cells (zone III) with quantification 3 days postinfection with Ruby-NS1 PR8 (500 PFU in 10 μl of PBS) ( n = 3 to 4). Scale bars, 200 μm. ( F ) Kinetics of viral load measured by MDCK plaque assay on nasal tissues of nonpregnant and pregnant Ifnar1 −/− mice (50 PFU; n = 4 to 7). ( G ) Kinetics of nasal il-17A expression by RT-qPCR (50 PFU; n = 4 to 5). ( H ) Representative FACS plots and frequency of γδ + T cells, ( I ) absolute number of γδ + T cells, ( J ) proportion of IL-17A–producing cells and ( K ) total number of IL-17A + γδ + T cells in nasal tissues of nonpregnant and pregnant mice at 0 and 1 day post-IAV infection following 4 hours of stimulation with rIL-23 (10 ng/ml) and rIL-1β (10 ng/ml) (50 PFU; n = 7 to 10). ( L ) Ns1 and ( M ) M1 expression in nasal tissues of WT and TCR δ −/− pregnant mice by RT-qPCR 3 days post-IAV infection (50 PFU; n = 5). ( N ) Ns1 and ( O ) M1 expression in nasal cavities of TCR δ −/− nonpregnant and pregnant mice 3 days post-IAV infection ( n = 5 to 8). Data produced from . Data are presented as mean ± SEM. with * P < 0.05, ** P < 0.01, and *** P < 0.001. Data were analyzed using two-way ANOVA followed by Sidak’s multiple comparisons [(B), (D), and (F) to (K)], unpaired Student’s t test [(E), (L), (N), and (O)], or Mann-Whitney U test (M).

Article Snippet: Nonpregnant mice were administered rIL-17A (1 μg per mouse; R&D Systems, #7956-ML-025/CF) intranasally in 10 μl of PBS (5 μl per naris) 2 hours before IAV infection (50 PFU intranasally).

Techniques: Infection, Plaque Assay, Fluorescence, Microscopy, Expressing, Quantitative RT-PCR, Produced, MANN-WHITNEY

Journal: Science Advances

Article Title: Pregnancy enhances antiviral immunity independent of type I IFN but dependent on IL-17–producing γδ + T cells in the nasal mucosa

doi: 10.1126/sciadv.ado7087

Figure Lengend Snippet: ( A to E ) Muc5ac , Cramp , Mbd-3 , Mbd-4 , and Mbd-14 , expression in the nasal cavities of nonpregnant and pregnant IAV-infected mice measured by RT-qPCR at various time points postinfection (50 PFU; n = 4 to 5). ( F ) Loading plot and correlation coefficients of Ns1 and AMP expression by RT-qPCR 3 days post-IAV infection ( n = 4 to 5). ( G ) Intraperitoneal (i.p.) administration of anti-IgG1 (αIgG1) or anti–IL-17A (αIL-17A) (200 μg per mouse in 200 μl of PBS) to nonpregnant and pregnant mice 2 days before and the day of IAV infection (50 PFU in 10 μl). Created with BioRender.com . ( H to K ) Quantification of nasal Muc5ac , Cramp , Mbd-3 , and Mbd-4 expression by RT-qPCR ( n = 10 to 11) and ( L ) nasal viral titers quantified by MDCK plaque assay 1 day post-IAV infection following intraperitoneal administration of αIL-17A ( n = 7 to 8). ( M to O ) Neutrophil, monocyte, and macrophage absolute numbers in the nasal cavities of nonpregnant and pregnant mice at days 0 and 1 post-IAV infection measured by flow cytometry (50 PFU; n = 7 to 11). Viral ( P ) Ns1 and ( Q ) M1 gene expression measured by RT-qPCR in murine epithelial cells 24 hours post-IAV infection (MOI of 1) with/without recombinant mBD-3 (rmBD-3; 100 μg/ml) ( n = 3). ( R ) Intranasal administration of rmBD-3 (1 μg per mouse; in 10 μl PBS) 1 hour post-IAV infection (50 PFU in 10 μl of PBS) to nonpregnant mice. Created with BioRender.com . Viral ( S ) Ns1 and ( T ) M1 gene expression measured by RT-qPCR in nasal tissues of nonpregnant mice 1 day post-IAV infection with/without rmBD-3 ( n = 7 to 8). Data are presented as mean ± SEM. with * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001. Data were analyzed using two-way ANOVA followed by Sidak’s multiple comparisons [(A) to (E) and (M) to (O)], Pearson’s correlation (F), unpaired Student’s t test [(H), (K), (L), (P), and (Q)], or Mann-Whitney U test [(I), (J), (S), and (T)].

Article Snippet: Nonpregnant mice were administered rIL-17A (1 μg per mouse; R&D Systems, #7956-ML-025/CF) intranasally in 10 μl of PBS (5 μl per naris) 2 hours before IAV infection (50 PFU intranasally).

Techniques: Expressing, Infection, Quantitative RT-PCR, Plaque Assay, Flow Cytometry, Recombinant, MANN-WHITNEY

Journal: Nature Communications

Article Title: Antibody Fc-receptor FcεR1γ stabilizes cell surface receptors in group 3 innate lymphoid cells and promotes anti-infection immunity

doi: 10.1038/s41467-024-50266-4

Figure Lengend Snippet: A Experimental scheme of C. rodentium infection model ( Methods ). B Representative periodic acid–Schiff (PAS) staining images of colons of Rorc-cre – Fcer1g f/f and Rorc-cre + Fcer1g f/f mice on day 7 post the infection as in ( A ). C Quantification of the histological scores of images as in ( B ) ( n = 5 mice per group). D, E Representative images of bacterial load determined by bioluminescent imaging in the colon ( D ) and the quantification ( E ) ( n = 6 mice per group). F-I Representative flow plots showing the intracellular abundance of IL-22 ( F ) or IL-17A ( H ) in total, CCR6 + and CCR6 – ILC3s from siLPs. Quantification of the frequencies of IL-22 ( G, Rorc-cre – Fcer1g f/f , n = 11 mice; Rorc-cre + Fcer1g f/f , n = 8 mice) or IL-17A ( I, Rorc-cre – Fcer1g f/f , n = 11 mice; Rorc-cre + Fcer1g f/f , n = 8 mice) -expressing cells in each compartment. Data are representative of two ( B-E, F, H ) or are pooled from two ( G, I ) independent experiments shown as the mean ± SEM. Statistical significance was tested by two-tailed t test ( C, E, G, I ).

Article Snippet: Mice were injected intraperitoneally with a dose of 1 μg rIL-17A protein (R&D Systems) or 0.8 μg rIL-22 protein (R&D Systems) 6 h before C. albicans infection, followed by a booster dose of the same cytokine at 24 h after infection.

Techniques: Infection, Staining, Imaging, Expressing, Two Tailed Test

Journal: Nature Communications

Article Title: Antibody Fc-receptor FcεR1γ stabilizes cell surface receptors in group 3 innate lymphoid cells and promotes anti-infection immunity

doi: 10.1038/s41467-024-50266-4

Figure Lengend Snippet: A Heatmap showing the expression (color bar, Z score) of genes (rows) significantly induced in ILC3s by the fungal infection (Supplementary Fig. ) across different genotypes and conditions (columns) ( Rorc-cre – Fcer1g f/f PBS, n = 4 mice; Rorc-cre + Fcer1g f/f C.A ., n = 4 mice; Rorc-cre – Fcer1g f/f C.A ., n = 3 mice). C.A . represents Candida albicans . B Quantification of the overall expression ( Methods ) of the genes in ( A ). C Quantification of total, CCR6 + and CCR6 – ILC3s in the siLPs of Rorc-cre − Fcer1g f/f and Rorc-cre + Fcer1g f/f mice on day 5 post the infection ( n = 6 mice per group). D-G Representative flow plots showing the intracellular abundance of IL-22 ( D ) or IL-17A ( F ) in total, CCR6 + and CCR6 – ILC3s on day 5 post the infection. Quantification of the frequencies of IL-22 ( E, Rorc-cre – Fcer1g f/f , n = 7 mice; Rorc-cre + Fcer1g f/f , n = 6 mice) or IL-17A ( G, Rorc-cre – Fcer1g f/f , n = 7 mice; Rorc-cre + Fcer1g f/f , n = 6 mice) -expressing cells in each compartment. H, I Representative flow plots showing the phosphorylation of JAK1, JAK2 and JAK3 in total ILC3s from siLPs of indicated mice on day 5 post the C. albicans infection ( H ). Quantification of MFI ( I, Rorc-cre – Fcer1g f/f , n = 5 mice; Rorc-cre + Fcer1g f/f , n = 4 mice). Data are representative of two ( C – G ) or one ( H , I ) or from two ( A , B ) independent experiments shown as the mean ± SEM. Statistical significance was tested by two-tailed t test ( B, C, E, G, I ).

Article Snippet: Mice were injected intraperitoneally with a dose of 1 μg rIL-17A protein (R&D Systems) or 0.8 μg rIL-22 protein (R&D Systems) 6 h before C. albicans infection, followed by a booster dose of the same cytokine at 24 h after infection.

Techniques: Expressing, Infection, Two Tailed Test

Journal: Nature Communications

Article Title: Antibody Fc-receptor FcεR1γ stabilizes cell surface receptors in group 3 innate lymphoid cells and promotes anti-infection immunity

doi: 10.1038/s41467-024-50266-4

Figure Lengend Snippet: A Experimental scheme. B Survival curves of C. albicans- infected Rorc-cre + Fcer1g f/f mice treated with IL-17A, IL-22 or vehicle (PBS) ( Rorc-cre – Fcer1g f/f PBS, n = 18 mice; Rorc-cre + Fcer1g f/f PBS, n = 21 mice; Rorc-cre + Fcer1g f/f IL-22, n = 13 mice; Rorc-cre + Fcer1g f/f IL-17A, n = 11 mice). C Quantification of the fungi load in kidneys of mice as in ( A ) on day 5 post the infection ( Rorc-cre – Fcer1g f/f PBS, n = 18 mice; Rorc-cre + Fcer1g f/f PBS, n = 16 mice; Rorc-cre + Fcer1g f/f IL-22, n = 7 mice; Rorc-cre + Fcer1g f/f IL-17A, n = 11 mice). D, E Representative PAS staining images of kidneys of mice as in ( A ) on day 5 post the infection. Boxes: medulla area with mycelia, showing in higher magnification in ( E ). F Quantification of IL-17A in the serum of mice as in Fig. on day 5 post the infection. n = 6 mice per group. Data are representative of three ( D, E ) or one ( F ) or are pooled from four ( B ) or three ( C ) independent experiments shown as the mean ± SEM. Statistical significance was tested by two-tailed t test ( C, F ) or log-rank (Mantel-Cox) test ( B ).

Article Snippet: Mice were injected intraperitoneally with a dose of 1 μg rIL-17A protein (R&D Systems) or 0.8 μg rIL-22 protein (R&D Systems) 6 h before C. albicans infection, followed by a booster dose of the same cytokine at 24 h after infection.

Techniques: Infection, Staining, Two Tailed Test

Journal: Nature Communications

Article Title: Antibody Fc-receptor FcεR1γ stabilizes cell surface receptors in group 3 innate lymphoid cells and promotes anti-infection immunity

doi: 10.1038/s41467-024-50266-4

Figure Lengend Snippet: A, B Representative flow plots showing the intracellular abundance of IL-17A ( A ) in total, CCR6 + and CCR6 – ILC3s from siLPs of mice with the indicated genotypes on day 5 post C. albicans infection. Quantification of the frequencies of IL-17A-expressing cells in each compartment ( B , WT, n = 7 mice; Ncr1 –/– , n = 5 mice; Fcgr3 –/– , n = 4 mice). C Survival curves of C. albicans- infected Rag1 −/− Rorc-cre − Fcer1g f/f ( n = 8 mice) and Rag1 −/− Rorc-cre + Fcer1g f/f mice ( n = 7 mice). All mice were intravenously infected with C. albicans on day 0 ( Methods ). D Survival curves of C. albicans- infected Rag1 −/− Rorc-cre − Fcer1g f/f and Rag1 −/− Rorc-cre + Fcer1g f/f mice treated with mIgG ( Methods ) ( n = 8 mice per group). E, F Representative flow plots showing the intracellular abundance of IL-17A ( E ) in total, CCR6 + and CCR6 – ILC3s from siLPs of mice in ( D ) on day 5 post the infection. Quantification of the frequencies of IL-17A ( F )-expressing cells in each compartment ( n = 4 mice per group). Data are pooled from three ( A, B ) or representative of three ( C ) or two ( D-F ) independent experiments shown as the mean ± SEM. Statistical significance was tested by two-tailed t test ( B, F ) or log-rank (Mantel-Cox) test ( C, D ).

Article Snippet: Mice were injected intraperitoneally with a dose of 1 μg rIL-17A protein (R&D Systems) or 0.8 μg rIL-22 protein (R&D Systems) 6 h before C. albicans infection, followed by a booster dose of the same cytokine at 24 h after infection.

Techniques: Infection, Expressing, Two Tailed Test