Journal: Cell reports

Article Title: Tuft-cell-intrinsic and -extrinsic mediators of norovirus tropism regulate viral immunity

doi: 10.1016/j.celrep.2022.111593

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Mouse IFN-λ2 , Peprotech , Cat# 250-33.

Techniques: Virus, Recombinant, Saline, Reverse Transcription, SYBR Green Assay, Software, Membrane

Journal: mBio

Article Title: IFN-λ drives distinct lung immune landscape changes and antiviral responses in human metapneumovirus infection

doi: 10.1128/mbio.00550-24

Figure Lengend Snippet: IFN-λ is highly upregulated during HMPV infection. Mice were infected with 5 × 10 5 PFU of HMPV strains TN/94-49 or C2-202 or mock cell lysate and IFN-λ2/3, IFN-β, and IFN-γ were measured in lung homogenate day 1 post-infection by (A) Luminex; ( B ) ELISA; or (C) qPCR. Data in C were normalized to HPRT1 gene and mock-infected mice by the 2 −∆∆ Ct method. ( D ) Mice were infected with HMPV C2-202 and IFN-λ2/3, IFN-β and IFN-γ were measured in lung homogenate days 1, 5, and 7 post-infection by Luminex. ( E ) BEAS-2B cells were infected with TN/94-49 or C2-202 and IFN-λ1 and IFN-λ2 measured in supernatant days 0–3 post-infection by Luminex. ( F ) C10 cells were infected with TN/94-49 or C2-202 and IFN-λ1 and IFN-λ2 measured in supernatant days 0–3 post-infection by Luminex. Cells were infected at multiplicity of infection (MOI) of 0.05. Limit of detection noted by dashed line. Data are shown as mean ± standard deviation. Analyses by student’s one-way or two-way ANOVA. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: For treatment, recombinant human IFN-λ1 protein (Peprotech; catalog #300-02), recombinant mouse IFN-β protein (Sino Biologicals; catalog #50708), or recombinant mouse IFN-λ2 protein (Preprotech; catalog #250-33) were reconstituted in 0.1% bovine serum albumin (BSA).

Techniques: Infection, Luminex, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: mBio

Article Title: IFN-λ drives distinct lung immune landscape changes and antiviral responses in human metapneumovirus infection

doi: 10.1128/mbio.00550-24

Figure Lengend Snippet: Inflammatory cytokine expression is altered by loss of IFN-λ signaling. WT (shown in black), Ifnar1 −/− (labeled IFNAR −/− , shown in green), and Ifnlr1 −/− (labeled IFNLR −/− , shown in purple) mice were infected with 5 × 10 5 PFU C2-202 HMPV. ( A ) Protein expression levels by Luminex assay (ng/mL) of inflammatory cytokines in lung homogenates collected on day 1 post-infection from 5 × 10 5 PFU C2-202 HMPV-infected mice graphically represented by heat map. ( B ) Protein expression levels by Luminex of IFN-λ2/3, IFN-β, IFN-γ, IL-6, MIP-1α, and MCP-1 cytokines in lung homogenate from WT, Ifnlr1 −/− (labeled IFNLR −/− ), and Ifnar1 −/− (labeled IFNAR −/− ) mice infected with 5 × 10 5 PFU C2-202 HMPV and harvested day 1 post-infection. Data are shown in 7A and 7B from one representative Luminex plate ( n = 3 mice per group) with similar findings on repeat assay. ( C ) qPCR analysis of interferon-stimulated genes was done for lung homogenate of mice described in 7B. Data are shown as mean ± standard deviation. Analyses were done by one-way ANOVA. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: For treatment, recombinant human IFN-λ1 protein (Peprotech; catalog #300-02), recombinant mouse IFN-β protein (Sino Biologicals; catalog #50708), or recombinant mouse IFN-λ2 protein (Preprotech; catalog #250-33) were reconstituted in 0.1% bovine serum albumin (BSA).

Techniques: Expressing, Labeling, Infection, Luminex, Standard Deviation

Journal: mBio

Article Title: IFN-λ drives distinct lung immune landscape changes and antiviral responses in human metapneumovirus infection

doi: 10.1128/mbio.00550-24

Figure Lengend Snippet: IFN-λ prophylaxis and treatment reduce HMPV burden without inflammation. C57BL/6 ( B6 ) mice were infected with 5 × 10 5 PFU C2-202 HMPV. ( A ) In a prophylaxis model (schematic diagram shown), recombinant IFN-λ (1 µg) or IFN-β (5 µg) was administered intranasally 1 days prior to infection. Mock prophylaxis included intranasal delivery of an equal volume of 0.1% bovine serum albumin (BSA). Mice receiving mock prophylaxis (shown in black), IFN-β prophylaxis (shown in green), and IFN-λ prophylaxis (shown in purple) were characterized ( B–F ). Disease was assessed by measuring body weight ( B ) and clinical severity scores ( C ) to day 7 post-infection. Weight represented as % of day 0. Clinical severity scores were measured by assigning 1 point out of 5 for each of the following criteria: hunching, huddling, fur ruffling, rapid breathing, and lethargy. Analyses were done by two-way ANOVA. ### P < 0.001, #### P < 0.0001 for mock vs both IFN-β and IFN-λ. ** P < 0.01, *** P < 0.001, **** P < 0.0001 for IFN-λ vs IFN-β. ( D, E ) HMPV titer (PFU/g) was measured in lung homogenates ( D ) or nasal turbinates ( E ) of mice receiving IFN prophylaxis. ( F ) Frequency of lung innate immune and epithelial cell populations in mice receiving IFN prophylaxis euthanized day 1 post-infection by flow cytometry. ( G ) In a treatment model (schematic diagram shown), recombinant IFN-λ (1 µg) or IFN-β (5 µg) was administered intranasally 2 days post-infection. Mice receiving mock treatment (shown in black), IFN-β treatment (shown in green), and IFN-λ treatment (shown in purple) were characterized ( G–K ). Disease was assessed by measuring body weight ( H ) and clinical severity scores ( I ) to day 7 post-infection. Mock treatment included intranasal delivery of an equal volume of 0.1% BSA. ( J, K ) HMPV titer (PFU/g) was measured in lung homogenates ( J ) or nasal turbinates ( K ) of mice receiving treatment. Limit of detection noted by dashed line. Analyses were done by one-way ANOVA. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: For treatment, recombinant human IFN-λ1 protein (Peprotech; catalog #300-02), recombinant mouse IFN-β protein (Sino Biologicals; catalog #50708), or recombinant mouse IFN-λ2 protein (Preprotech; catalog #250-33) were reconstituted in 0.1% bovine serum albumin (BSA).

Techniques: Infection, Recombinant, Flow Cytometry