Journal: medRxiv

Article Title: Puumala orthohantavirus dysregulates hyaluronan metabolism in lung cells and correlates with disease severity and lung impairment

doi: 10.64898/2026.03.10.26348053

Figure Lengend Snippet: (A) Schematic overview of the human lung cell lines used in the study. (B–F) PUUV infection kinetics over five days, quantified as the percentage of infected cells by immunofluorescence staining for PUUV nucleocapsid protein with nuclear counterstaining in (B) A549 alveolar epithelial cells, (C) BEAS-2B bronchial epithelial cells, (D) HULEC-5a microvascular lung endothelial cells, (E) MRC-5 lung fibroblasts, and (F) A549/PIV5-V cells with impaired STAT1 signaling. Statistical significance was calculated by unpaired t-test (*P < 0.05, **P < 0.01; ns, not significant). (G–K) HA levels in culture media supernatants collected at the indicated time points from PUUV-infected (G) A549, (H) BEAS-2B, (I) HULEC-5a, (J) MRC-5, and (K) A549/PIV5-V cells. HA concentrations were measured by ELISA and are shown as percentage of mock-infected controls. Data represents three independent experiments with duplicate technical replicates per experiment. Bars show mean ± SEM. Statistical significance was assessed using multiple unpaired t-tests with Holm–Šídák correction for multiple comparisons (*P < 0.05, **P < 0.01, ***P < 0.001; ns, not significant).

Article Snippet: Human lung microvascular endothelial cells HULEC-5a (CRL-3244TM, American Type Culture Collection, Manassas, VA, USA) were cultured in MCDB 131 medium (Sigma-Aldrich, St. Louis, MO, USA) supplemented with 10 ng/mL human epidermal growth factor (Sigma-Aldrich, St. Louis, MO, USA), 1 μg/mL hydrocortisone (Sigma-Aldrich, St. Louis, MO, USA), 100 U/mL Penicillin-Streptomycin and 10% FBS.

Techniques: Infection, Immunofluorescence, Staining, Enzyme-linked Immunosorbent Assay

Journal: medRxiv

Article Title: Puumala orthohantavirus dysregulates hyaluronan metabolism in lung cells and correlates with disease severity and lung impairment

doi: 10.64898/2026.03.10.26348053

Figure Lengend Snippet: (A) Schematic overview of genes involved in HA synthesis ( HAS1–3 ), HA degradation ( HYAL1–2 ), and HA signaling ( CD44 ). (B–F) Expression of HA-regulating genes in (B) A549, (C) BEAS-2B, (D) HULEC-5a, (E) MRC-5, and (F) A549/PIV5-V cells, determined by quantitative PCR at the indicated time points. Expression levels were normalized to β-actin and are shown as fold change relative to uninfected controls. Data represents three independent experiments with duplicate technical replicates per experiment. Results are displayed as box-and-whisker plots, where the box indicates the 25th to 75th percentiles (interquartile range), the line within the box represents the median, and the whiskers denote the minimum and maximum values. Statistical significance was assessed using paired t-tests, comparing the expression values to the respective time-matched uninfected controls (*P < 0.05, **P < 0.01, ***P < 0.001).

Article Snippet: Human lung microvascular endothelial cells HULEC-5a (CRL-3244TM, American Type Culture Collection, Manassas, VA, USA) were cultured in MCDB 131 medium (Sigma-Aldrich, St. Louis, MO, USA) supplemented with 10 ng/mL human epidermal growth factor (Sigma-Aldrich, St. Louis, MO, USA), 1 μg/mL hydrocortisone (Sigma-Aldrich, St. Louis, MO, USA), 100 U/mL Penicillin-Streptomycin and 10% FBS.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Whisker Assay