Journal: bioRxiv
Article Title: IFNγ-Driven Inflammatory Responses in the Nasal Mucosa Drive Influenza Virus Shedding and Transmission
doi: 10.64898/2026.01.06.698034
Figure Lengend Snippet: ( A ) IFNγ supplementation in rPR8-H3N2-infected pups alters shedding dynamics . C57BL/6J (WT) or IFNγ⁻/⁻ index pups were intranasally infected with 300 PFU of rPR8-H3N2 and cohoused with uninfected contacts for 4 days. Index pups received intranasal IFNγ supplementation (1 ng/μL) or vehicle control beginning 4–6 hours post-infection and daily thereafter. Viral titers from day 4 URT lavage (left), cumulative shedding from days 1–4 (middle), and day 4 URT titers from contact pups (right) were quantified by plaque assay. ( B ) IFNγ-dependent transcriptional responses in the URT . Expression of selected antiviral and inflammatory genes in URT cellular fractions was quantified by qRT–PCR at 4 dpi. ( C ) IFNγ restores mucus production in IFNγ⁻/⁻ mice . URT secretions from IFNγ⁻/⁻ pups infected with rPR8-H3N2 and treated intranasally with IFNγ or vehicle control were collected at 2 dpi. URT lavages collected at 2 dpi were dotted on nitrocellulose and probed with anti-Muc5AC antibody. Signal intensity was normalized to positive control, and fold-change over mock is shown. Median values are indicated. ( D ) IFNγ induces mucus secretion in the absence of infection . URT secretions from uninfected pups treated intranasally with escalating doses of IFNγ (10 ng/μL, 1 ng/μL, 0.1 ng/μL) or vehicle, as well as from rPR8-H3N2-infected pups, were collected at 2 dpi. URT lavages collected at 2 dpi were dotted on nitrocellulose and probed with anti-Muc5AC antibody. Signal intensity was normalized to positive control, and fold-change over mock is shown. Median values are indicated. Statistical differences among groups were assessed using the Kruskal–Wallis test. ( E ) IFNγ enhances shedding during rPR8-H1 infection . Index pups infected with 300 PFU of rPR8-H1 received intranasal IFNγ (1 ng/μL/g), IFNβ (1 ng/μL/g) or vehicle beginning 4–6 hpi. Viral titers from day 4 URT lavage (left), cumulative shedding from days 1–4 (middle), and day 4 URT titers from contact pups (right) were quantified by plaque assay. Statistics . Differences between two groups were analyzed using the Mann–Whitney test. Differences among multi-group medians were analyzed by Kruskal–Wallis test. Experiments represent ≥2 independent biological replicates. Significance is denoted as *P < 0.05. Cartoons created with BioRender.com; graphs generated using GraphPad Prism 10.
Article Snippet: Pups were treated intranasally with IFNγ (R&D Systems, Lot #169049915, Minneapolis, MN, USA) or IFNβ (R&D Systems, #8234-MB-010, Minneapolis, MN, USA) at 10-0.1ng/μL diluted in 0.1% PBS/BSA and intranasally daily for the duration indicated.
Techniques: Infection, Control, Plaque Assay, Expressing, Quantitative RT-PCR, Positive Control, MANN-WHITNEY, Generated
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