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[Construction, expression and immunogenicity of EV71 multiepitope-mGITRL eukaryotic plasmid]
Zhen-wei Mao1, Hai-bing Liu, Dong Zheng
1Department of Laboratory Medicine, Affiliated People's Hospital of Jiangsu University, Zhenjiang, China. maopen365@163.com
Summary
This study constructed a novel enterovirus 71 (EV71) multiepitope-mGITRL DNA vaccine. The vaccine successfully expressed fusion protein and induced a significant anti-EV71 immune response in mice.
Area of Science:
- Molecular Biology
- Immunology
- Vaccine Development
Background:
- Enterovirus 71 (EV71) is a significant pathogen causing hand, foot, and mouth disease.
- Developing effective EV71 vaccines is crucial for public health.
- Multiepitope strategies combined with immune enhancers show promise for vaccine design.
Purpose of the Study:
- To construct an EV71 multiepitope-mGITRL eukaryotic plasmid.
- To evaluate the immunogenicity of the constructed plasmid in BALB/c mice.
Main Methods:
- Designed and synthesized a VP1' epigene with B and T cell epitopes.
- Amplified and cloned the mGITRL gene into an expression vector.
- Constructed the pIRES-VP1'-mGITRL recombination plasmid and transfected it into COS7 cells.
- Detected protein expression via Western blotting and measured antibody titers using ELISA in immunized mice.
Main Results:
- Successfully constructed and sequenced the pIRES-VP1'-mGITRL plasmid.
- Confirmed high expression of VP1'-mGITRL fusion protein in COS7 and muscle cells.
- Demonstrated a high titer of anti-VP1 antibody in the serum of immunized BALB/c mice.
Conclusions:
- The pIRES-VP1'-mGITRL plasmid facilitates high expression of VP1'-mGITRL fusion protein.
- This construct effectively elicits a robust immune response in mice, indicating its potential as an EV71 vaccine candidate.
