Co-immunization with two recombinant Newcastle disease viruses expressing ILTV glycoprotein B and H9N2 AIV HA confers cross-protection against three avian pathogens

File 1: Viral growth kinetics in BHK-21 and CEF 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. File 2: The original image of Western blot analysis.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. File 3: Protective efficacy in SPF chicks. Antibody titers against NDV, AIV and ILTV after immunization, and the date of survival rate, clinical symptoms, tissue viral load and virus shedding after challenging with NDV, AIV and ILTV in SPF chicks File 4: Protective efficacy in commercial chicks. Antibody titers against NDV, AIV and ILTV after immunization, and the date of survival rate, clinical symptoms, tissue viral load and virus shedding after challenging with NDV, AIV and ILTV in commercial chicksFile 1: BHK-21与CEF细胞的病毒生长动力学实验。将细胞以感染复数（Multiplicity of Infection, MOI）0.01分别感染或共感染rTS-H9/rTS-gB；每隔12小时收集细胞培养上清液，通过间接免疫荧光试验（Indirect Immunofluorescence Assay, IFA）在指定细胞系中测定病毒滴度。 File 2: 蛋白质印迹（Western Blot）分析原始图像。将BHK-21或CEF细胞以感染复数（Multiplicity of Infection, MOI）0.1分别感染或共感染rTS-H9/rTS-gB；于感染后48小时（hours post infection, hpi）收集细胞裂解物，分别对BHK-21细胞（泳道F）与CEF细胞（泳道G）进行蛋白质印迹分析。使用针对HA（H9N2亚型禽流感病毒，H9N2 AIV）、gB（传染性喉气管炎病毒，ILTV）以及新城疫病毒核蛋白（NDV NP）的特异性抗体进行蛋白检测，以小鼠抗肌动蛋白多克隆抗体作为上样对照。 File 3: 无特定病原体（Specific Pathogen Free, SPF）雏鸡的免疫保护效力实验。包含免疫接种后针对新城疫病毒（Newcastle Disease Virus, NDV）、禽流感病毒（Avian Influenza Virus, AIV）与传染性喉气管炎病毒（Infectious Laryngotracheitis Virus, ILTV）的抗体滴度，以及SPF雏鸡经NDV、AIV、ILTV攻毒后的存活率、临床症状、组织病毒载量与病毒排毒量相关数据。 File 4: 商品代雏鸡的免疫保护效力实验。包含免疫接种后针对新城疫病毒（Newcastle Disease Virus, NDV）、禽流感病毒（Avian Influenza Virus, AIV）与传染性喉气管炎病毒（Infectious Laryngotracheitis Virus, ILTV）的抗体滴度，以及商品代雏鸡经NDV、AIV、ILTV攻毒后的存活率、临床症状、组织病毒载量与病毒排毒量相关数据。