Additional Insertion of gC Gene Triggers Better Immune Efficacy of TK/gI/gE-Deleted Pseudorabies Virus in Mice.

In recent years, pseudorabies virus (PRV) variants have resulted in an epidemic in swine herds and huge economic losses in China. Therefore, it is essential to develop an efficacious vaccine against the spread of PRV variants. Here, the triple-gene-deletion virus and the triple-gene-deletion plus gC virus were constructed by homologous recombination (HR). And then, their growth capacity, proliferation ability, and immune efficacy were evaluated. The results showed that the growth kinetics of the recombinant viruses were similar to those of the parental strain PRV-AH. Compared with the triple-gene-deletion virus group, the more dominant level of neutralizing antibody (NA) can be induced in the triple-gene-deletion plus gC virus group with the same 106.0 TCID50 dose after 4 and 6 weeks post-initial immunization (PII) (p < 0.0001). In addition, the antibody titers in mice immunized with the triple-gene-deletion plus gC virus were significantly higher than those immunized with triple-gene deletion virus with the same 105.0 TCID50 dose after 6 weeks PII (p < 0.001). More importantly, in the triple-gene-deletion plus gC virus group with 105.0 TCID50, the level of NA was close to that in the triple-gene deletion virus group with 106.0 TCID50 at 6 weeks PII. Meanwhile, the cytokines IL-4 and IFN-γ in sera were tested by enzyme-linked immunosorbent assay (ELISA) in each group. The highest level of IL-4 or IFN-γ was also elicited in the triple-gene deletion plus gC virus group at a dose of 106.0 TCID50. After challenge with PRV-AH, the survival rates of the triple-gene deletion plus gC virus immunized groups were higher than those of other groups. In immunized groups with 105.0 TCID50, the survival rate shows a significant difference between the triple-gene deletion plus gC virus group (75%, 6/8) and the triple-gene deletion virus group (12.5%, 1/8). In general, the immune efficacy of the PRV TK/gI/gE-deleted virus can be increased with additional gC insertion in mice, which has potential for developing an attenuated vaccine candidate for PRV control.

The Role of Latency-Associated Transcripts in the Latent Infection of Pseudorabies Virus.

Pseudorabies virus (PRV) can cause neurological, respiratory, and reproductive diseases in pigs and establish lifelong latent infection in the peripheral nervous system (PNS). Latent infection is a typical feature of PRV, which brings great difficulties to the prevention, control, and eradication of pseudorabies. The integral mechanism of latent infection is still unclear. Latency-associated transcripts (LAT) gene is the only transcriptional region during latent infection of PRV which plays the key role in regulating viral latent infection and inhibiting apoptosis. Here, we review the characteristics of PRV latent infection and the transcriptional characteristics of the LAT gene. We also analyzed the function of non-coding RNA (ncRNA) produced by the LAT gene and its importance in latent infection. Furthermore, we provided possible strategies to solve the problem of latent infection of virulent PRV strains in the host. In short, the detailed mechanism of PRV latent infection needs to be further studied and elucidated.

Better immune efficacy triggered by the inactivated gI/gE-deleted pseudorabies virus with the additional insertion of gC gene in mice and weaned pigs.

A new variant of pseudorabies virus (PRV) with high pathogenicity has been prevalent in many swineherds vaccinated with Bartha-K61 in China since 2011. Several gene-deleted vaccine candidates have been developed based on new emerging PRV variants. PRV-AH, a new emerging PRV strain from Anhui Province, was isolated in our laboratory in 2013. In the present study, rPRV-AH-gI-/gE- and rPRV-AH-gI-/gE-/gC+ were generated based on PRV-AH by homologous recombination. The growth kinetics of rPRV-AH-gI-/gE- and rPRV-AH-gI-/gE-/gC+ were similar to their parental strains. Compared with the commercial inactivated vaccine of Ea strain, the immune efficacy of the inactivated vaccine based on recombinant viruses was evaluated in mice and weaned pigs. The result showed that the level of neutralizing antibody in mice immunized with rPRV-AH-gI-/gE-/gC+ was higher compared with those immunized with rPRV-AH-gI-/gE- at a dose of 106 TCID50 at 8 weeks post initial immunization (p < 0.0001). Among the groups immunized at a dose of 105 TCID50, the rPRV-AH-gI-/gE- group showed a survival rate of 37.5 %, while the rPRV-AH-gI-/gE-/gC+ group showed a protection rate of 87.5 % against the PRV-AH challenge. Besides, the rPRV-AH-gI-/gE- and rPRV-AH-gI-/gE-/gC+ group immunized at a dose of 106 TCID50 showed a survival rate of 100 %. Interestingly, compared with the commercial vaccine group, the group of 105 TCID50 rPRV-AH-gI-/gE-/gC+ showed a lower level of neutralizing antibodies (p < 0.0001) but the same protection rate in mice. Moreover, in the pig experiment, the level of neutralizing antibodies in the group vaccinated with inactivated rPRV-AH-gI-/gE-/gC+ was higher than any other groups at 8 weeks post initial immunization (p < 0.05). More importantly, the milder symptoms and pathological lesions occurred in pigs vaccinated with rPRV-AH-gI-/gE-/gC+ after challenge with 106 TCID50 PRV-AH, revealing that additional insertion of gC gene could enhance the protective efficacy in PRV gI/gE-deleted vaccine in pigs. Collectively, these above-mentioned findings suggested that the inactivated vaccine of rPRV-AH-gI-/gE-/gC+ had a better immune efficacy, which could be regarded as a promising inactivated vaccine candidate for PRV control.