PMC:7115396 / 33527-38958 JSONTXT

{"target":"http://pubannotation.org/docs/sourcedb/PMC/sourceid/7115396","sourcedb":"PMC","sourceid":"7115396","source_url":"https://www.ncbi.nlm.nih.gov/pmc/7115396","text":"The spike protein is the coronavirus structural proteins, which plays an important role on pathogenicity in coronaviruses [6], [50]. However, replacing the S gene of Beaudette strain by the virulent strains does not make it virulent [51], [52]. The S protein can be cleaved into S1 and S2 subunits at the S1/S2 cleavage site [53], the S1 subunit is responsible for receptor binding to the host cells [54]. Whether the S1 gene has an effect on the virulence of the virus is unknown. In this study, the recombinant strain, rH120-S1/YZ, expressed the S1 gene of the QX-like strain, which retains the S1/S2 cleavage site of H120, maintained the ability to replicate and exhibited pathogenicity in embryos. However, providing IBYZ S1 sequences to H120 did not increase its virulence to that of rIBYZ. IBVs replicate in many epithelial cells, including respiratory tissues, the alimentary canal, kidney, gonads, and bursa [2], [6]. The QX-like rIBYZ strain is capable of considerable growth in the trachea, lung, and kidney, which leads to severe tissue damage. In addition, this strain caused cilia necrosis and loss, lung congestion and hemorrhage, nephritis, as well as inflammatory cell infiltration in these tissues in infected chickens at different time points post-infection. The ability of IBV to replicate within many respiratory, enteric, and other epithelial surfaces may be partially related to the fact that the attachment of IBV to host cells is dependent on sialic acid on the cell surface being recognized by the S1 subunit [6], [54], [55]. However, the binding of virus to the host cell is the first step in determining tropism and S2 is responsible for membrane fusion [56]. Thus, to further explore the different tissue tropism between rH120-S1/YZ and rIBYZ, we examined the viral loads and histopathology in the tissues of the rH120-S1/YZ-infected group. It was found that rH120-S1/YZ remains moderately pathogenic in the trachea (e.g., some cilia necrosis and loss and decreased secretory cells) but no lesions were observed in the lungs and kidneys of infected chickens, in contrast to chickens infected with the rIBYZ strain. This finding suggests that both the S2 and S1 genes might play an important role in viral tissue tropism as also suggested by others [57]. Another possibility is that the lack of viral replication and damage in kidney following rH120-S1/YZ is due to lower replication ability of backbone except S2 gene from its parental rH120 strain. The viral load of the respiratory-type strain, rH120 in the different tissues indicated that the viral levels in the trachea, early following infection were significantly higher than those of rIBYZ, which indicates that the ability of the virus to attached to trachea epithelial cells of the rH120 strain was stronger than that of the rIBYZ strain. However, during the later time points post-infection, the level of the rH120 strain was significantly lower than rIBYZ, which suggests that the virulence of IBV depends both on tissue invasion and viral replication ability, as well as other functions mediated by other viral proteins (e.g., non-structural, structural, and accessory proteins) [6], [29], [58], [59], [60]. The reduction of the viral load at 5 dpi in the group infected with the rIBYZ strain may be associated with the necrosis and shedding of the trachea epithelium. Moreover, the ability to replication in the different organs of chickens infected with rH120-S1/YZ as reflected by viral loads at multiple time points was lower than that exhibited in the chickens infected with the rIBYZ strain, which demonstrated that the S1 subunit was not the only determinant of viral tropism that was consistent with results by others [57], [61], [62], and the significant difference in viral load was related to differences in the viral backbone. However, when comparing the rH120-S1/YZ and rH120 strains which carry the same backbone, it was found that the viral RNA of rH120-S1/YZ in the lungs and kidneys was significantly lower than that of rH120, which may due to the replacement of the IBYZ S1 gene in the recombinant virus, whose structure on the surface contains some degree of changes, which we speculated that may affects the functionality of the S protein of coronaviruses. Although cryo-EM structure of IBV spike protein has been determined, the relation between structure and function of different IBV S1 genes is unknown. [63]. In addition to the growth in epithelial tissues observed during the early infection period, IBV can establish long-term persistent infections in chicken flocks via oral and cloacal shedding, which represents a substantial challenge to IBV control [1], [6]. In this study, we detected viral RNA in the oral and cloacal swabs isolated from infected chickens at 7, 14, 18, 22, and 26 dpi by real-time PCR. Compared with rH120 and rIBYZ, the viral load in both the oral and cloacal swabs of the rH120-S1/YZ group at different time points after infection was significantly decreased. However, while it has been reported that the persistent virus will be re-excreted at the point of lay [1], the persistence of rH120-S1/YZ strain in the egg-laying chickens needs to be further verified. In a word, due to its low virulence i presented in virus loading and lesion in different tissue, oral and cloacal shedding of host animals, rH120-S1/YZ can be considered a safe attenuated vaccine candidate for young 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