Porcine reproductive and respiratory syndrome virus (PRRSV) is a major viral threat to swine, causing significant economic loss in the global pig farming industry. This virus includes two major genotypes, PRRSV1 and PRRSV2, both characterized by high mutation rates and genetic variability, complicating the development of a universally effective vaccine and disease control. To address this challenge, this study utilizes immunoinformatics tools to identify conserved epitopes and design a multi-epitope vaccine candidate against PRRSV based on reverse vaccinology. The complete sequences of PRRSV-encoded proteins were retrieved worldwide, and the conserved fragments were identified through the alignment of polypeptide sequences. Subsequent screening was conducted to screen epitopes for their potential to be safe and to activate B cells, HTLs (helper T cells), and CTLs (cytotoxic T cells). By conjugating the selected epitopes with distinct adjuvant proteins, three vaccine candidates were designed and termed PRRSV-vaccine (PRRSV-V-1, PRRSV-V-2, and PRRSV-V-3, respectively). Furthermore, systematic evaluations of their physicochemical properties, structural stability, binding with pattern recognition receptors, and induction of the host immune system were performed. PRRSV-V-2 had the most promising physicochemical and structural characteristics, strong binding with toll-like receptors (TLR3 and TLR8), and the most vigorous reactions to host immune responses. As the most promising candidate, the recombinant PRRSV plasmid was in silico designed for expression in Escherichia coli. Our study proposed a novel approach to PRRSV vaccine development against PRRSV, offering a promising strategy for controlling the infection across diverse PRRSV strains in swine. Despite providing significant insights into vaccine design through computational methods, the results of this study remain predictive. So, it is open for the experimental validations of the scientific community to ensure its actual immunological properties, especially the safety and efficacy.

Background: Laryngeal cancer (LC) is characterized by high mortality and remains challenging in prognostic evaluation and treatment benefits. Ubiquitin-related genes (UbRGs) are widely involved in cancer initiation and progression, but their potential value in LC is unknown.Methods: RNA-seq and clinical data of LC were obtained from TCGA and GEO. UbRGs that independently influenced the overall survival (OS) of LC patients were screened with differential expression, COX and LASSO regression analyses. A prognostic signature was then established and assessed for its predictive value, stability and applicability using Kaplan-Meier analysis and receiver operating characteristic curves. The nomogram was further generated in combination with the signature and clinical characteristics. Characterization of immune properties and prediction of drug sensitivity were investigated on the signature-based subgroups using a panel of in silico platforms. Verification of gene expression was conducted with Western blot, qRT-PCR and ELISA, ultimately.Results: PPARG, LCK and LHX1 were identified and employed to construct the UbRGs-based prognostic signature, showing a strong ability to discriminate LC patients with distinct OS in TCGA-LC and GSE65858, and excellent applicability in most clinical conditions. The nomogram showed higher predictive value and net clinical benefit than traditional indicators. As evaluated, the low-risk group had a more activated immune function, higher infiltration of anti-cancer immune cells and stronger expression of immune-promoting cytokines than the high-risk group. Immune properties were also correlated with individual signature genes. PPARG and LHX1 were negatively correlated, whereas LCK positively correlated, with the immuno-promoting microenvironment. Additionally, chemotherapy would be more effective in high-risk patients, while immune checkpoint inhibitors would be more effective in low-risk patients. Finally, dysregulation of the signature genes was confirmed in LC cell lines by Western blot, and PPARG knockdown significantly reduced the expression of the immunosuppressive cytokines IL6, TGFB1, TGFB2 and VEGFC by qRT-PCR and ELISA.Conclusion: We have developed a UbRGs-based signature for LC prognostic evaluation that is valuable in clinical application, indicative of the immune microenvironment and beneficial for individualized treatment guidance.

Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) is a major viral threat to swine, causing a significant economic loss in the global pig farming industry. This virus includes two major genotypes, both characterized by high mutation rates and genetic variability, complicating the development of a universally effective vaccine and the control of disease consequently. To address this challenge, a novel multi-epitope-based vaccine was designed to elicit a robust and cross-protective immune response against multiple PRRSV strains. The complete sequences of PRRSV-encoded proteins were retrieved worldwide, and the conserved fragments were identified through the alignment of polypeptide sequences. Subsequent screening was conducted screened epitopes for their potential to be safe and to activate B cells, CTLs, and HTLs. By conjugating the selected epitopes with distinct adjuvant proteins, three vaccine candidates were designed and termed as PRRSV-Vaccine (PRRSV-V-1, PRRSV-V-2, and PRRSV-V-3 respectively). Furthermore, systematic evaluations of their physiochemistry properties, structural stability, binding with pattern recognition receptors, and induction of the host immune system were performed. PRRSV-V-2 was demonstrated with the most promising physicochemical and structural characteristics, strong binding with TLR3 and TLR8, and the most vigorous reactions of host immune response. As the most promising candidate, the recombinant PRRSV plasmid was in-silico designed for expression in Escherichia coli. Our study proposed a novel approach to PRRSV vaccine development against PRRSV, offering a promising strategy for controlling the infection across diverse PRRSV virus strains in swine.

Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) is a major viral threat to swine, causing significant economic loss in global pig farming industry. The virus exists in two major genotypes, both characterized by high mutation rates and genetic variability, complicating the development of a universally effective vaccine and the control of disease consequently. To address this challenge, a novel multi-epitope-based vaccine was designed to elicit a robust and cross-protective immune response against multiple PRRSV strains. The complete sequences of PRRSV encoded proteins were retrieved worldwide, and the conserved fragments were identified through alignment of polypeptide sequences. Subsequent screening was conducted screened epitopes for their potential to be safe and to activate B cells, CTLs and HTLs. By conjugating the selected epitopes with distinct adjuvant proteins, three vaccine candidates were constructed and termed as PRRSV-Vaccine (PRRSV-V-1, PRRSV-V-2, and PRRSV-V-3 respectively). Furthermore, systematic evaluations of their physiochemistry properties, structural stability, binding with pattern recognition receptors and induction of host immune system were performed. PRRSV-V-2 was demonstrated with most promising physicochemical and structural characteristics, strong binding with TLR3 and TLR8, and most vigorous reactions of host immune response. As the most promising candidate, the recombinant PRRSV plasmid was in-silico designed for expression in Escherichia coli. Our study proposed a novel approach to PRRSV vaccine development against PRRSV, offering a promising strategy for controlling the infection across diverse PRRSV virus strains in swine.