Abstract
Since 2015, the outbreaks of hydropericardium-hepatitis syndrome (HHS) and inclusion body hepatitis (IBH) caused by the highly pathogenic serotype 4 fowl adenovirus (FAdV-4) and serotype 8 fowl adenovirus (FAdV-8), respectively, have caused huge economic losses to the poultry industry. Although several vaccines have been developed to control HHS or IBH, a recombinant genetic engineering vaccine against both FAdV-4 and FAdV-8 has not been reported. In this study, recombinant FAdV-4 expressing the fiber of FAdV-8b, designated as FA4-F8b, expressing fiber of FAdV-8b was generated by the CRISPR-Cas9 and homologous recombinant techniques. Infection studies in vitro and in vivo revealed that the FA4-F8b replicated efficiently in LMH cells and was also highly pathogenic to 2-week-old SPF chickens. Moreover, the inoculation of inactivated the FA4-F8b in chickens could not only induce highly neutralizing antibodies, but also provide efficient protection against both FAdV-4 and FAdV-8b. All these demonstrate that the inactivated recombinant FA4-F8b generated here can act as a vaccine candidate to control HHS and IBH, and FAdV-4 can be an efficient vaccine vector to deliver foreign antigens.
Highlights
Fowl adenovirus (FAdV) belongs to the family Adenoviridea and genus Aviadenovirus [1].Based on the profile of restriction enzyme digestion and sera cross-neutralization assay, FAdV is clustered into five species (FAdV-A~E) with 12 serotypes (FAdV-1~8a,8b~11) [2–4]
To generate a recombinant virus FAdV-4 carrying the fiber of FAdV-8b, two sgRNAs targeting C- and N-terminus of EGFP and fiber-2 of the FA4-EGFP, respectively, were first designed and cloned into lentiCRISPR v2, and the donor plasmid containing the fiber of FAdV-8b and fiber-2 of FAdV-4 was constructed
HPS and inclusion body hepatitis (IBH) caused by FAdV-4 and FAdV-8 have spread globally and resulted in huge economic losses to the poultry industry [8,11,24]
Summary
Fowl adenovirus (FAdV) belongs to the family Adenoviridea and genus Aviadenovirus [1]. Based on the profile of restriction enzyme digestion and sera cross-neutralization assay, FAdV is clustered into five species (FAdV-A~E) with 12 serotypes (FAdV-1~8a,8b~11) [2–4]. Epidemiological studies show that FAdV has spread worldwide and cause huge economic loss to the poultry industry. Chickens infected with FAdV generally show subclinical symptoms, while the acute infection of FAdV mainly causes inclusion body hepatitis (IBH), hepatitis-pericardial effusion syndrome (HHS) and gizzard erosion and ulceration (GEU) [3,5,6]. Clinical data indicate that different species or serotypes of FAdV can induce distinct pathogenic symptoms. HHS is mainly caused by the highly pathogenic
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