Journal: Transboundary and Emerging Diseases

Article Title: Influenza A Virus Utilizes the Nasolacrimal System to Establish Respiratory Infection after Ocular Exposure in the Swine Model

doi: 10.1155/2024/8192499

Figure Lengend Snippet: Ex vivo porcine conjunctiva culture supports the productive IAV infection. Ex vivo cultures of conjunctiva were incubated with H1N1 or H9N2 at 5 × 10 3 TCID 50 . The tissues were collected for IHC staining or western blot analysis and the supernatants were used to perform plaque assay at different time points. (a) Viral antigens were detected in the ex vivo culture using IHC. Positive cells were stained brown. Bars: 20 μ m. (b) Viral titer in culture supernatant of ex vivo culture was determined by plaque assay on MDCK. Each error bar represents the mean and SD of three independent experiments. (c) Protein expression of H1N1 HA-protein in ex vivo culture at different time points detected by western blotting. The intensity (HA/ β -actin) of H1N1-HA (left) and H9N2-HA (right) was measured using ImageJ. All data shown are the mean results ± SD from three independent experiments. Statistical significance was using one-way ANOVA. NS, no significance. ∗ P < 0.05, ∗∗ P < 0.01.

Article Snippet: After blocking with 5% skim milk, the membranes were explored with mouse anti-H1N1-HA monoclonal antibody (homemade) or rabbit anti-H9N2-HA antibody (Sino Biological, 11229-V08H, China). β -actin-HRP conjugated (Abways, AB2001, China) was used as the internal control to evaluate the protein amounts by using ImageJ.

Techniques: Ex Vivo, Infection, Incubation, Immunohistochemistry, Western Blot, Plaque Assay, Staining, Expressing

Journal: Nature Communications

Article Title: Characterization of two non-competing antibodies to influenza H3N2 hemagglutinin stem reveals its evolving antigenicity

doi: 10.1038/s41467-025-65595-1

Figure Lengend Snippet: A The binding activities of AG2-G02 and 2F02 against recombinant HA proteins from the specified influenza strains were measured by ELISA. The absorbance values at 450 nm are shown as a heatmap. B , C The neutralization activity of AG2-G02 and 2F02 against recombinant H3N8 and different recombinant H3N2 viruses was measured by a microneutralization assay, with antibodies ( B ) continuously present in the medium throughout the experiment, or ( C ) removed at one-hour post-infection (hpi). MIC: minimal inhibitory concentration. Each bar represents the mean of two independent biological replicates. Each data point represents one replicate. A–C Strain names are abbreviated as follows: H3N2 A/Hong Kong/1/1968 (HK/1968), H3N2 A/Philippines/2/1982 (Philippines/1982), H3N2 A/Beijing/109/1992 (Beijing/1992), H3N2 A/Wuhan/359/1995 (Wuhan/1995), H3N2 A/Moscow/10/1999 (Moscow/1999), H3N2 A/New York/55/2004 (NY/2004), H3N2 A/Wisconsin/67/2005 (Wisconsin/2005), H3N2 A/Uruguay/716/2007 (Uruguay/2007), H3N2 A/Switzerland/9715293/2013 (Switzerland/2013), H3N2 A/Darwin/6/2021 (Darwin/2021), H3N8 A/mallard/Alberta/362/2017 (Alberta/2017), H4N6 A/mallard/Alberta/455/2015 (Alberta/2015), H7N3 A/Canada/rv444/2004 (Canada/2004), H7N9 A/Hong Kong/125/2017 (HK/2017), H7N9 A/Anhui/1/2013 (Anhui/2013), H14N5 A/mallard/Astrakhan/263/1982 (Astra/1982), and H15N2 A/Australian shelduck/Western Australia/1756/1983 (WAus/1983).

Article Snippet: HA proteins of the following strains were purchased from SinoBiological: H3N2 A/HongKong/1/1968, H3N2 A/Beijing/109/1992, H3N2 A/Wuhan/359/1995, H3N2 A/Moscow/10/1999, H3N2 A/Switzerland/9715293/2013, H14N5 A/mallard/Astrakhan/263/1982, and H15N2 A/Australian shelduck/Western Australia/1756/1983.

Techniques: Binding Assay, Recombinant, Enzyme-linked Immunosorbent Assay, Neutralization, Activity Assay, Microneutralization Assay, Infection, Concentration Assay, Western Blot

Journal: Nature Communications

Article Title: Characterization of two non-competing antibodies to influenza H3N2 hemagglutinin stem reveals its evolving antigenicity

doi: 10.1038/s41467-025-65595-1

Figure Lengend Snippet: Female BALB/c mice at six weeks old were injected intraperitoneally with 5 mg/kg of the indicated antibody four hours prior to challenge with 5 × LD 50 of H3N2 Philippines/1982 (X-79). CR9114 was used as a positive control for protection. A Kaplan-Meier survival curves are presented ( n = 7 for control, n = 5 for experimental groups). B The mean percentage of body weight change post-infection is shown ( n = 7 for the control group, n = 5 for each experimental group). The humane endpoint was defined as a 25% weight loss from the initial weight on day 0. Data are presented as mean ± standard deviation. C Lung viral titers on day 3 post-infection are shown ( n = 3 per group). Each bar represents the mean. Each data point represents one mouse.

Article Snippet: HA proteins of the following strains were purchased from SinoBiological: H3N2 A/HongKong/1/1968, H3N2 A/Beijing/109/1992, H3N2 A/Wuhan/359/1995, H3N2 A/Moscow/10/1999, H3N2 A/Switzerland/9715293/2013, H14N5 A/mallard/Astrakhan/263/1982, and H15N2 A/Australian shelduck/Western Australia/1756/1983.

Techniques: Injection, Positive Control, Control, Infection, Standard Deviation

Journal: Nature Communications

Article Title: Characterization of two non-competing antibodies to influenza H3N2 hemagglutinin stem reveals its evolving antigenicity

doi: 10.1038/s41467-025-65595-1

Figure Lengend Snippet: A Occurrence of different amino acid variants at HA2 position 32 of human H3N2 strains from 1968 to the present was analyzed based on 33,000 human H3N2 HA sequences from the GSAID database . The x-axis represents the years, while the y-axis shows the percentage of strains containing a particular amino acid at HA2 position 32 in a given year. Thr, Ile, and Arg are represented by orange, brown, and green lines, respectively. B The structures of AG2-G02 binding to HA stem with I32 HA2 and R32 HA2 are modeled. The binding affinity ( K d ) of AG2-G02 Fab to different HA stem variants is indicated in parentheses. C–E The binding of plasma samples from ( C ) pre-2003 adults ( n = 20), ( D ) post-2011 adults ( n = 20), and ( E ) post-2011 infants ( n = 15) to H3 stem with either T32 HA2 , I32 HA2 or R32 HA2 is measured by ELISA. The y-axis represents the area under the curve (AUC) of four serial 10-fold dilutions of serum (1:100, 1:1,000, 1:10,000, and 1:100,000). P -values were determined using a paired two-tailed Student’s t -test.

Article Snippet: HA proteins of the following strains were purchased from SinoBiological: H3N2 A/HongKong/1/1968, H3N2 A/Beijing/109/1992, H3N2 A/Wuhan/359/1995, H3N2 A/Moscow/10/1999, H3N2 A/Switzerland/9715293/2013, H14N5 A/mallard/Astrakhan/263/1982, and H15N2 A/Australian shelduck/Western Australia/1756/1983.

Techniques: Binding Assay, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Cell reports

Article Title: Memory B Cell Activation, Broad Anti-influenza Antibodies, and Bystander Activation Revealed by Single-Cell Transcriptomics

doi: 10.1016/j.celrep.2019.12.063

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Influenza A H9N2 (A/Chicken/Hong Kong/G9/97) Hemagglutinin / HA Protein (His Tag) , Sino Biological , 40036-V08H.

Techniques: Recombinant, Multiplex Assay, Cell Isolation, Sequencing, Software