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Despite the robust induction of humoral immune responses, a limitation of many adjuvants is their weak stimulation of cellular immunity. The development of synthetic gene-encoding adjuvants for simultaneous induction of both humoral and cell-mediated immune responses is under study. In this study, we examined the impact of toll/interleukin-1 receptor (TIR) domain of toll-like receptor 7 (TLR7) as molecular adjuvants on potency of inactivated infectious bursal disease (IBD) vaccines. A total of 60 specific pathogen-free week-old chicks were randomized grouped to receive either TIR-TLR7-adjuvanted IBD-inactivated vaccine or inactivated IBD antigen along with an unvaccinated control. Serum antibody titers were measured to estimate the humoral immunity, as well as lymphocyte proliferation activity for cellular immune responses. The protection was estimated after challenge with a very virulent IBD virus (IBDV) strain at 4 weeks postvaccination. The results indicated that one dose of IBD/TIR-TLR7 vaccine induced specific antibody responses, whereas a lower response after administration of inactivated IBD antigen was observed. The stimulation of splenocytes results indicated that the TIR-TLR7 adjuvanted IBD vaccine is capable of modulating cell-mediated immune response in treated chickens. A full protection against IBDV infection was achieved by injection of one dose IBD/TIR-TLR7 vaccine in the challenge trial. This study demonstrated that codelivery of TIR-TLR7 with inactivated IBD antigen resulted in simultaneous enhancing immune responses against IBD. [ABSTRACT FROM AUTHOR]

Infectious laryngotracheitis virus (ILTV) infection causes economic losses in the poultry industry. Outbreaks of the infection in commercial poultry are controlled by using attenuated live vaccines. The genomic and antigenic characteristics between virulent and vaccine strains of ILTV are very similar, thus the differential diagnosis of ILTV strains is targeted in controlling programs. Here, the chicken embryonic neural stem (cENS) cell serves as a host model to study virulence characteristics of ILTVs. The susceptibility of cENS cells to different ILTV strains infection was investigated by analyzing the impact of infection on cell death, virus latency, and determining the host cell responses. We found that cENS cells are highly susceptible to vaccine strain ILTV infection. Upon infection, the strain showed faster replication kinetics in cell culture and marked cytopathic effects with affecting the expression of cellular neuropeptide Y (NPY). Virulent strain was able to enter cENS cells, but no infectious virus was produced at third passage. The establishment of latency state was confirmed by molecular detection of viral ICP4 gene and virus reactivation assay. In contrast to vaccine strain, transcription level of cellular NPY was also upregulated significantly following virulent strain infection. The difference expression pattern of NPY in ILTV strains-infected cells is most closely with the presence of CPE in culture. The neural stem cell system may provide the most relevant tool for differential diagnosis of ILTV strains. [ABSTRACT FROM AUTHOR]

The infectious bursal disease virus (IBDV) causes an acute and highly contagious immunosuppressive response in young chickens by targeting B lymphocytes in immune organs. Changes in regulatory T-cell ratio and apoptosis have been demonstrated during IBDV infection in these cells. The possible change in CD19 expression as the precursor of B cells after IBDV replication was detected in this study. Raji cells were infected with an IBDV isolate at MOIs of 1.0 and 3.0. The viral kinetics were determined using the characteristic virus-induced CPE, cell viability, and infectious titer. Induction of apoptosis and also changes in the CD19 expression within the virus infection were assessed by flow cytometry. The Raji cells were found to be susceptible to IBDV infection by producing marked CPEs dependent on MOI. The infectivity titers were determined in intra- and extracellular samples at the defined hours. The kinetics of early IBDV replication in Raji cells were nearly identical for both MOIs, but a significant difference in the infectivity titer was observed at 48 hpi. The quick apoptotic events were observed to be significantly higher in MOI 3.0, which was correlated with the lower virus titer. A significant CD19 expression change in the IBDV-infected Raji cells was revealed. The results suggested that Raji cells mimic the IBDV replication in lymphoid organs and the virus replication is related to CD19 expression frequencies in the lymphoid cells. • IBDV targets the medullary bursa in the avian host with a preference for B cells. • Raji cell line has the potential for IBDV replication and support higher viral growth titers. • Changes in Bregs expression be suitable in monitoring IBDV replication in a B cell-originated cell line. • The change in the level of CD19 marker is closely associated with IBDV infection. [ABSTRACT FROM AUTHOR]

• The most IL-6 and TNF-α gene expression was observed in full flagellin adjuvanted group. • The recombinant full flagellin adjuvanted group showed the highest cellular activity. • The highest HI titer was observed in commercial vaccine and recombinant flagellin adjuvanted groups. The present study was designed to investigate the potential application of native (N) and recombinant (truncated modified [tmFliC] and full-length [flFliC]) flagellin proteins along with inactivated Newcastle disease virus (NDV). Fifty six SPF chickens were immunized twice with PBS (control), inactivated NDV (Ag), inactivated NDV/flFliC (AgF), inactivated NDV/tmFliC (AgT), inactivated NDV/N (AgN), commercial vaccine containing Montanide (Vac) and Vac/N (VacN), with a two-week interval. Blood was collected weekly and spleens were harvested after chickens were sacrificed. Interleukin-6 (IL-6) and tumor necrotic factor-α (TNF-α) gene expression in peripheral blood mononuclear cells were analyzed by Real-Time PCR. Antibody response was assessed by haemagglutination inhibition (HI). Cellular activity was quantified by MTT assay. Results showed that the most IL-6 and TNF-α gene expression was observed in AgF group (P < 0.01). The lowest gene expression among vaccinated groups was observed in Ag group for IL-6 and Ag and Vac group for TNF-α. The highest HI titer was observed in Vac, VacN, AgF and AgT groups. The AgF group showed the highest cellular activity (P < 0.01). In conclusion, flagellin-adjuvanted groups showed a pro-inflammatory effect and acted similarly to or better than the Vac group. Hence, flagellin can be proposed as a potential adjuvant for ND vaccine. [ABSTRACT FROM AUTHOR]