A Plant-Produced Recombinant Fusion Protein-Based Newcastle Disease Subunit Vaccine and Rapid Differential Diagnosis Platform

Fanshu Ma,Jiquan Ou,Heng Yin,Erqin Zhang,Xiangyue Zhao,Shenli Zhang,Yanan Wang,Ruiguang Deng,Kunpeng Li,Enmin Zhou,Xiangxiang Niu,Qingmei Li,Li Wang,Gaiping Zhang,Xueyang Li,Qianru Xu

doi:10.3390/vaccines8010122

Abstract

Newcastle disease (ND) is a highly contagious avian disease, causing considerable economic losses to the poultry industry. To obtain a safe, inexpensive, and effective ND vaccine to meet the international trade requirements of differentiating infected from vaccinated animals (DIVA), here we report the production of Oryza sativa recombinant fusion (F) protein in stably transformed transgenic rice seeds via agroinfiltration. The F protein expression level was enhanced 3.6-fold with a genetic background in low glutelin. Inoculation of plant-produced F antigen into Specific Pathogen Free (SPF) chickens markedly elicited neutralizing antibody responses against homologous and heterologous ND virus strains. Two doses of 4.5 μg fully protected chickens from a lethal ND challenge without any clinical symptoms. The mean weight gain of F protein-immunized chickens within 15 days after challenge was significantly higher than that of traditional whole virus vaccine-immunized chickens, thereby obtaining higher economic benefits. Moreover, the sera from the chickens vaccinated with the plant-produced F vaccine did not show reactivity in an immunochromatographic strip targeting the haemagglutinin-neuraminidase protein (HN) protein, and DIVA could be achieved within 10 minutes. Our results demonstrate that the plant-derived F vaccine along with immunochromatographic strips could be useful in the implementation of an NDV eradication program.

Highlights

Newcastle disease (ND), an acute and highly contagious avian disease caused by avian paramyxovirus, remains endemic throughout Africa, Asia, and Europe, with frequent epidemics occurring despite the availability of global commercial vaccinations since the 1930s [1,2]
To obtain high-level expression of the recombinant haemagglutinin-neuraminidase protein (HN)-dimer in rice seed, the F gene sequence was synthesized using rice-preferred codons followed by the introduction of a strong endosperm-specific promoter Gtl3a and its signal peptide to target the F protein into the protein storage vacuoles
To determine the expression of NDV-F in transgenic seeds, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot analysis revealed a predominant protein band of approximately 55 kDa in transgenic grains, which was not detected in the TP309 control rice seeds (Figure 1d)

Summary

Introduction

Newcastle disease (ND), an acute and highly contagious avian disease caused by avian paramyxovirus, remains endemic throughout Africa, Asia, and Europe, with frequent epidemics occurring despite the availability of global commercial vaccinations since the 1930s [1,2]. Vaccines 2020, 8, 122 virus is introduced into a susceptible flock, including domestic poultry, virtually all the birds will be infected within two to six days (World Organisation for Animal Health, OIE, 2019). According to the World Animal Health Information Database (WAHIS), an average of 60 countries reported. ND continues to be a threat and to cause considerable economic losses to the poultry industry worldwide. ND in its highly pathogenic form is listed in the World Organization for Animal Health (OIE) Terrestrial Animal. Due to the constant threat of introduction of the virus from wild birds, vaccination and the establishment of poultry biosecurity are essential to the prevention of ND. ND vaccines are available in live, inactivated forms and are licensed for use in many countries worldwide

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