PEVB Viral Vector Delivery for Stable Dual-Tract Pig Vaccines
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Solution Overview
Problem
There is a lack of efficient viral vector delivery systems for vaccines targeting the respiratory and digestive tracts of pigs, particularly for diseases caused by pathogens like African swine fever virus (ASFV), which affects the economy and animal health.
Innovation Solution
A viral vector delivery system is developed using a backbone plasmid with a full-length cDNA of porcine enterovirus B (PEVB) and a helper plasmid with a green fluorescent protein-coding gene, constructed through PCR and co-transfection, achieving high efficiency and stability, with a viral titer of 10^7.75 TCID50/mL.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If viral vectors are used to deliver vaccines to respiratory and digestive tracts of pigs, then immunization effect is improved, but system complexity increases
Solution Approach 1:
The delivery system is segmented into two functional plasmids: a backbone plasmid containing the viral genome and a helper plasmid containing the T7 RNA polymerase gene. This segmentation allows each plasmid to perform a specific function, simplifying the overall system design while maintaining high immunization effectiveness through coordinated action of the segmented components.
Solution Approach 2:
The viral vector system is designed with multi-functionality to serve as both a delivery vehicle and an expression system. The plasmids are engineered to simultaneously achieve viral replication, antigen expression, and delivery to respiratory and digestive tract tissues, reducing the need for separate systems and lowering overall complexity.
2Productivity
If viral titer is increased to improve delivery efficiency, then gene delivery efficiency is improved, but system stability deteriorates
Solution Approach 1:
The system optimizes viral titer by adjusting key parameters including the promoter strength in the backbone plasmid, the ratio of backbone to helper plasmid during transfection, and the transfection conditions. These parameter changes enable achievement of high viral titer (10^7.75 TCID50/mL) while maintaining system stability through controlled optimization rather than extreme conditions.
3Reliability
If pathogenic epitopes are inserted to enhance vaccine effectiveness, then immunization effect is improved, but pathogenicity increases
Solution Approach 1:
The system converts potentially harmful pathogenic epitopes into beneficial vaccine antigens by inserting them into a recombinant viral vector that has been engineered to be replication-deficient or attenuated. The epitopes from pathogens like ASFV, PEDV, and PDCOV are expressed in a controlled manner that stimulates immune response without causing the harmful diseases associated with the original pathogens.
Data Source
AI summary
A viral vector delivery system for both respiratory and digestive tracts of pigs and an application thereof are provided. The viral vector delivery system includes a backbone plasmid and a helper plasmid. The backbone plasmid is produced by inserting a full-length cDNA of porcine enterovirus B (PEVB) into a pUC57 plasmid. The helper plasmid is produced by inserting a green fluorescent protein-coding gene into a plasmid pCAG-T7-polymerase. The viral vector delivery system can be constructed with high efficiency, can quickly cause a cytopathic effect (CPE) after infecting cells, has a viral titer up to 107.75 TCID50/mL, and can maintain high stability. A pathogenic epitope for the respiratory and digestive tracts of pigs is inserted into the backbone plasmid to produce a vaccine antigen, which is non-pathogenic, has high stability and a high viral titer, and allows a prominent immunization effect after being inoculated in pigs.