Journal: Frontiers in Immunology

Article Title: Comparison of air-liquid interface transwell and airway organoid models for human respiratory virus infection studies

doi: 10.3389/fimmu.2025.1532144

Figure Lengend Snippet: Comparison of influenza A/H1N1 virus infection susceptibility and infection kinetics of airway air-liquid interface (ALI) transwell and organoid models. (A) At 24 hpi, infected ALI transwell cultures were immunostained to detect viral nucleoprotein (NP, orange), ciliated cells (α-tubulin, green), and actin filaments (phalloidin, purple). Scale bar equals 50 μm. (B, C) At 24 hpi, infected organoids cultured in Matrigel (B) or BME2 (C) were visualized by immunostaining of formalin-fixed and paraffin-embedded sections (for Matrigel) and non-paraffin-embedded organoids (for BME2) to detect viral nucleoprotein (NP, red), actin filaments were stained with pan-cytokeratin (PAN-CK, green) or phalloidin (green), and nuclei were stained by DAPI (blue). Scale bars equal 100 μm. (D–F) Virus quantification was performed by TCID50 assay on Madin-Darby Canine Kidney (MDCK) cells for all 3 models. TCID50/ml for the time points 2, 24, 48, and 72 hpi are shown. (D) At the indicated time points, apical washes were collected from infected ALI transwell cultures (MOI=0.01) from 3 donors (A/B/C). The data shown are from 3 technical replicates (inserts) per donor. (E) Human airway organoids derived from 3 donors (D/E/F) were cultured in Matrigel, infected following enzymatic dissociation (MOI=0.1), and sampled at the indicated hpi. The data shown are from 2 technical replicates (wells) per donor. (F) Organoids derived from 3 donors (G/H/I) were cultured in BME2, infected with influenza virus following mechanical disruption (MOI=0.01), and sampled at the indicated hpi. The data shown are from 1 (I) or 2 (G/H) technical replicates (wells) per donor. Replicates were obtained in 2 independent experiments. The limit of detection was 2.89 TCID50/mL for panel (D) and 3.16 TCID50/mL for panels (E, F) . The geometric mean with geometric standard deviation (SD) is shown.

Article Snippet: Samples were stained with the primary Influenza A H1N1 Nucleoprotein Antibody (1:2000; Novus Biologicals, #NBP2-16965, USA).

Techniques: Comparison, Virus, Infection, Cell Culture, Immunostaining, Staining, TCID50 Assay, Derivative Assay, Disruption, Standard Deviation

Journal: Frontiers in Immunology

Article Title: Comparison of air-liquid interface transwell and airway organoid models for human respiratory virus infection studies

doi: 10.3389/fimmu.2025.1532144

Figure Lengend Snippet: Dynamics of host responses to A/H1N1 influenza virus infection in air-liquid interface (ALI) transwell and airway organoid models, measured by cytokine production. Concentrations of the cytokines CXCL10, IL-6, IL-8, IFN-λ1, IFN-λ2/3, and IFN-β were measured using LEGENDplex assay. (A) Basolateral medium samples from (mock-)infected ALI transwell cultures of 3 donors (A/B/C) were collected at the indicated time points and assessed for cytokine production. The data shown are from 3 technical replicates (inserts) per donor. (B) Cytokine concentrations were measured in the supernatant of (mock-)infected dissociated airway organoids cultured in Matrigel, derived from 3 donors (D/E/F). The data shown are from 2 technical replicates (wells) per donor. (C) Cytokine concentrations were measured in the supernatant of (mock-)infected dissociated airway organoids cultured in BME2 from 3 donors (G/H/I). The data shown are from 1 (I) or 2 (G/H) technical replicates (wells) per donor. Replicates were obtained in 2 independent experiments. The geometric mean with geometric SD is shown. For visualization purposes, cytokine concentrations below the limit of quantification (LOQ) were assigned values of 0.5 times the LOQ. Dotted lines and grey shading indicate the LOQ for each cytokine.

Article Snippet: Samples were stained with the primary Influenza A H1N1 Nucleoprotein Antibody (1:2000; Novus Biologicals, #NBP2-16965, USA).

Techniques: Virus, Infection, Cell Culture, Derivative Assay

Journal: RSC Medicinal Chemistry

Article Title: De novo design of a stapled peptide targeting SARS-CoV-2 spike protein receptor-binding domain †

doi: 10.1039/d3md00222e

Figure Lengend Snippet: Binding kinetics study of pep39 and the spike protein RBD. (A) The deflection curve for the peptide pep39 immobilized on the cantilever in response to different concentrations of SARS-CoV-2 spike protein. Anti-S1 antibody served as a positive control and H1N1 protein, as a negative control. (B) The association and dissociation between immobilized spike protein RBD and different concentrations of pep39. Recombinant human ACE2 protein used as a positive control and PBS buffer as a reference blank.

Article Snippet: We purchased silicon cantilevers from Nanoworld Inc., and the SARS-CoV-2 spike protein RBD, the antibody for the spike protein RBD and influenza H1N1 hemagglutinin protein were procured from Sino Biological Inc; and 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide and sulfo-NHS from ThermoFisher Scientific.

Techniques: Binding Assay, Positive Control, Negative Control, Recombinant

Journal: Nature Communications

Article Title: Human antibody 3E1 targets the HA stem region of H1N1 and H5N6 influenza A viruses

doi: 10.1038/ncomms13577

Figure Lengend Snippet: ( a , b ) Prophylactic efficacy of 3E1 mAb against a lethal challenge with the A/Sichuan/09/2009(H1N1) virus ( a ) or A/CK/NX/S6/2014(H5N6) virus ( b ). The weight loss (left) and survival curves (right) of mice treated with 30, 10, 3 or 1 mg kg −1 of 3E1 mAb or PBS buffer 24 h before lethal challenge by an intranasal inoculation with the H1-SC09 or H5-NX14 virus (at day 0) are shown. ( c , d ) Therapeutic efficacy of 3E1 mAb against a lethal challenge with the A/Sichuan/09/2009(H1N1) ( c ) or A/CK/NX/S6/2014(H5N6) virus ( d ). The weight loss (left) and survival curves (right) of mice treated with PBS buffer (at day 1) or 20 mg kg −1 of 3E1 mAb right after or 1, 2, 3 days after lethal challenge by an intranasal inoculation with the H1-SC09 or H5-NX14 virus (at day 0) are shown. Error bars represent mean±s.d. Data represent a representative experiment from three independent experiments.

Article Snippet: The HAs from the A/Brevig Mission/1/1918(H1N1), A/WSN/1933(H1N1), A/Brisbane/59/2007(H1N1), A/California/07/2009(H1N1), A/Canada/720/2005(H2N2), A/Hong Kong/483/1997(H5N1), A/Vietnam/1203/2004(H5N1), A/Anhui/1/2005(H5N1), A/Sichuan/26221/2014(H5N6), A/Hong Kong/1073/1999(H9N2), A/black-headed gull/Netherlands/1/00(H13N8), A/black-headed gull/Sweden/5/99(H16N3), A/Hong Kong/1/1968(H3N2), A/Aichi/2/1968(H3N2), A/Brisbane/10/2007(H3N2), A/Taiwan/70113/2007(H3N2), A/Netherlands/219/2003(H7N7) and A/Zhejiang/DTID-ZJU01/2013(H7N9) viruses were purchased from Sino Biological Inc. HAs of the A/Washington/05/2011(H1N1) and A/California/04/2009(H1N1) viruses were prepared as described above.

Techniques: