h1n1 pr8  (ATCC)

Journal: iScience

Article Title: Ambient temperature regulates CD4 + T cell tonic T cell receptor signaling and responsiveness

doi: 10.1016/j.isci.2026.115801

Figure Lengend Snippet: Thermoneutrality-driven exacerbation of inflammatory disease severity is linked with increased CD4 + T cell TNF production (A–D) 8-to-10-week-old WT C57BL/6J male mice were fed a high-fat diet (HFD) and housed in Tn- or Ts-conditions for 20 weeks (n = 9–22 biological replicates per group). (A) Schematic overview. (B-D) The following parameters were quantified at the time of terminal harvest: Total body weight (B); serum alanine transaminase (ALT) levels via calorimetric assay (C); and frequency of TNF + hepatic CD4 + T cells (TCRβ + TCRγδ − NK1.1 − CD4 + ) upon ex vivo stimulation with phorbol 12-myristate 13-acetate (PMA; 50 ng/mL) and ionomycin (1 mg/mL) via flow cytometry (D). (E–H) 8-to-10-week-old WT C57BL/6J male mice were housed in Tn- or Ts-conditions. After 2 weeks, mice were intranasally infected with influenza virus (Charles River Influenza A/PR/8/34 [H1N1], Batch: 4XP160913; 30 Hemagglutinin units/mice; n = 5–7 biological replicates per group). (E) Schematic overview. (F) Body weight change post-infection. (G, H) The following parameters were quantified 5 days post-infection: bronchioalveolar lavage fluid (BALF) IL-6 levels via ELISA (G); and frequency of TNF + pulmonary CD4 + T cells (CD45 + TCRβ + TCRγδ − NK1.1 − CD8 − ) upon ex vivo stimulation with PMA (50 ng/mL) and ionomycin (1 mg/mL) via flow cytometry (H). (I-L) 12-week-old WT C57BL/6J male mice were housed in Tn- or Ts-conditions. After 2 weeks, mice were intratracheally treated with house dust mite (HDM; 10 μg/mouse) on days 0, 14, and 21 (n = 8–11 biological replicates per group). (I) Schematic overview. (J–L) The following parameters were quantified 3 days after final HDM exposure: BALF eosinophil count via morphological analysis (J); IL-5 production of single lung cell suspensions after 72-h ex vivo cell culture (K); and frequency of TNF + pulmonary CD4 + T cells (CD45 + TCRβ + TCRγδ − NK1.1 − CD8 − ) upon ex vivo stimulation with PMA (50 ng/mL) and ionomycin (1 mg/mL) via flow cytometry (L). (B-D, F-H, J-L) Mann-Whitney U test. ∗: p < 0.05, ∗∗∗: p < 0.001, and ∗∗∗∗: p < 0.0001. For bar and line graphs, data represent mean ± SEM.

Article Snippet: Mice were sedated with 2.5% isoflurane (Akorn, NDC 59399-106-01) and treated intranasally with 30 HA Units/7.16 log 10 EID/5.7 log 10 TCID50 H1N1 PR8 influenza (Charles River; Influenza A/PR/8/34, Batch: 4XP160913; provided by Dr. William Zacharias, CCHMC) resuspended in 0.9% sodium chloride solution (Hospira, NDC 0409-4888-02).

Techniques: Ex Vivo, Flow Cytometry, Infection, Virus, Enzyme-linked Immunosorbent Assay, Cell Culture, MANN-WHITNEY

Journal: bioRxiv

Article Title: S1PR1 signaling biases neutrophils toward long-lived low-inflammatory functional states

doi: 10.64898/2026.02.05.703783

Figure Lengend Snippet: (A) Schematic of high-dose H1N1 (LD 50 ) model over 21 days. (B) Survival curve of LysM -S1pr1 TG (n = 20) and control (n = 18) mice in response to H1N1 infection. Survival curves were compared using the Gehan–Breslow–Wilcoxon test. (C) Body weight loss of LysM -S1pr1 TG (n = 8) and control (n = 8) mice. (D) Blood oxygen saturation (SpO 2 ) in LysM -S1pr1 TG (n = 9) and control (n = 11) mice post-infection. Both (C) and (D) were tested by two-way mixed-effects analysis with Geisser–Greenhouse correction and Šídák’s multiple comparisons test; * p < 0.05, ** p < 0.01. (E) Schematic of the low-dose (0.16 LD 50 ) H1N1 model at 3 and 7 dpi. (F) Total cell count in BALF at 3 and 7 dpi (n = 7 mice per group). (G and H) BALF neutrophil and macrophage count at 3 and 7 dpi (n = 7 mice per group). (I-K) Expression of viral genes, including nucleoprotein (NP), polymerase acidic protein (PA), and hemagglutinin (HA), in the infected lung in control (n = 3) and LysM -S1pr1 TG (n = 4) mice. (L) Total protein in BALF in control (n = 3) and LysM -S1pr1 TG (n = 6) mice at 7 dpi. (M) IL-6 (ng/mL) secretion in BALF in control (n ≥ 4) and LysM- S1pr1 TG (n ≥ 5) mice at 3 and 7 dpi. (N) IL-10 (ng/mL) secretion in BALF in control (n ≥ 4) and LysM -S1pr1 TG (n ≥ 3) mice at 3 and 7 dpi. Data are mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001 (two-tailed unpaired Student’s t test).

Article Snippet: Influenza A/Puerto Rico 8/1934 (PR8) H1N1 strain was obtained from Charles River (Cat# 10100374, Wilmington, MA), aliquoted, and stored in liquid nitrogen.

Techniques: Control, Infection, Cell Characterization, Expressing, Two Tailed Test

Journal: bioRxiv

Article Title: S1PR1 signaling biases neutrophils toward long-lived low-inflammatory functional states

doi: 10.64898/2026.02.05.703783

Figure Lengend Snippet: (A) Kaplan–Meier survival curves of Mrp8- S1pr1 transgenic (TG) mice ( n = 25) and Cre-negative littermate controls ( n = 21) following intranasal infection with high-dose H1N1 (LD 50 ). Survival was monitored for up to 21 days post-infection. Statistical significance was assessed using the log-rank (Mantel–Cox) and Gehan–Breslow–Wilcoxon tests. (B) Body weight change in control (n = 13) and Mrp8- S1pr1 TG (n = 21) mice following influenza infection, expressed as a percentage of the initial body weight. Data represent mean ± SD. (C) Arterial oxygen saturation (SpO₂) measured longitudinally after infection using pulse oximetry in control (n = 13) and Mrp8- S1pr1 TG (n = 21) mice. Data represent mean ± SD. Statistical significance is denoted as * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Influenza A/Puerto Rico 8/1934 (PR8) H1N1 strain was obtained from Charles River (Cat# 10100374, Wilmington, MA), aliquoted, and stored in liquid nitrogen.

Techniques: Transgenic Assay, Infection, Control