Journal: Poultry Science

Article Title: Co-immunization with two recombinant Newcastle disease viruses expressing ILTV gB and H9N2 AIV HA confers protective efficacy against three avian pathogens

doi: 10.1016/j.psj.2026.106661

Figure Lengend Snippet: Generation and coinfection of rTS-H9 and rTS-gB in vitro . (A) Schematic diagram depicting the rNDV cDNA clones rTS-H9 and rTS-gB. (B, C) Viral growth kinetics in BHK-21 (B) and CEF (C) cells. The cells were coinfected or individually infected with rTS-H9/rTS-gB at an MOI of 0.01. Viral titers in culture supernatants collected at 12-hour intervals were determined by IFA on the indicated cell lines. For rTS-H9 + rTS-gB: the proliferation of rTS-H9 was detected using an anti-HA primary antibody. For rTS-gB + rTS-H9: the proliferation of rTS-gB was assessed via an anti-gB primary antibody. (D, E) Foreign protein expression analysis by IFA in BHK-21 (D) and CEF (E) cells. The cells were coinfected or individually infected with rTS-H9 and/or the rTS-gB mixture at an MOI of 0.01. At 48 hpi, the cells were fixed and stained with chicken anti-HA (H9N2) and rabbit anti-gB (ILTV) antibodies, followed by anti-chicken IgG (FITC, green) and rabbit anti-IgG (Cy3, red) antibodies. The nuclei were counterstained with DAPI (blue). Scale bar = 20 μm. (F, G) BHK-21 or CEF cells were coinfected or individually infected with rTS-H9/rTS-gB at an MOI of 0.1. Western blot analysis of BHK-21 (F) and CEF (G) cell lysates harvested at 48 hpi. Proteins were detected using specific antibodies against HA (H9N2 AIV), gB (ILTV), and NDV nucleoprotein (NP). The mouse anti-actin polyclonal antibody served as a loading control.

Article Snippet: Subsequently, fluorescein isothiocyanate (FITC)-conjugated anti-chicken IgG (1:200. bs0310R-FITC, Bioss) and Cy3-labeled anti-rabbit IgG (1:500.

Techniques: In Vitro, Clone Assay, Infection, Expressing, Staining, Western Blot, Control