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