Related Experiment Video
Updated: Jan 10, 2026
Optimization of Antigen Preparation for Hemagglutination Inhibition Assay of Newcastle Disease Virus Serology
Published on: January 24, 2025
[Construction of DNA vaccines containing C3d-P29 against Newcastle disease]
Xinxing Chu1, Jun Peng, Lixue Weng
1College of Animal Science and Veterinary Medicine, Shandong Agricultural University, Taian 271018, China. xxchu@sdau.edu.cn
Abstract:
After cloning the C3d cDNA of AA broilers using the liver mRNA source, a pair of primers were designed to subclone the P29 gene to the pUC19 plasmid. Several tandems of P29 were constructed in the pUC19 plasmid using a pair of isoschizomers-BamH I and Bgl II. The pUC- P29.n was igested to get the gene of P29.n that was then cloned to pCDNA3.1 (+) plasmid. After this, the F Gene of Newcastle Disease Virus was cloned through RT-PCR and inserted into the upstream of the P29.n that was in the pCDNA-P29.n, and the DNA vaccines containing F gene against NDV with C3d-P29 as molecular adjuvant were constructed. Several groups of Specefic Pathogen Free chickens were injected with these recombinant plasmids. The pCDNA-F-P29.4 and pCDNA-F-P29.6 group had higher HI antibody titers than the pCDNA-F group. The pCDNA-F-P29.4 and pCDNA-F-P29.6 group's HI antibody titers did not achieve titers as high as the inactive vaccine group. However, they all provided protection against the lethal F48E9 virus challenge.
Insights
New DNA vaccines for Newcastle Disease Virus (NDV) were created using C3d-P29 as a molecular adjuvant. These vaccines demonstrated protection against lethal NDV challenge in chickens, with enhanced antibody titers observed.
Area of Science:
- Molecular Biology
- Immunology
- Veterinary Science
Context:
- Newcastle disease virus (NDV) poses a significant threat to poultry health and global food security.
- Developing effective and safe vaccines is crucial for controlling NDV outbreaks.
- The C3d protein and P29 gene show potential as molecular adjuvants to enhance vaccine efficacy.
Purpose:
- To construct and evaluate DNA vaccines expressing the NDV F gene with C3d-P29 as a molecular adjuvant.
- To assess the immunogenicity and protective efficacy of these novel DNA vaccines in chickens.
Summary:
- The C3d cDNA of AA broilers was cloned, and the P29 gene was subcloned into plasmids.
- Tandems of P29 were constructed and cloned into pCDNA3.1(+).
- The NDV F gene was inserted upstream of P29.n, creating DNA vaccines (pCDNA-F-P29.n).
- Chickens vaccinated with pCDNA-F-P29.4 and pCDNA-F-P29.6 showed higher HI antibody titers than the control group (pCDNA-F).
- While antibody titers did not reach those of inactivated vaccines, all constructed vaccines provided protection against lethal NDV F48E9 virus challenge.
Impact:
- The study demonstrates the potential of C3d-P29 as a molecular adjuvant in DNA vaccines against NDV.
- These findings contribute to the development of improved vaccine strategies for poultry.
- The research offers a promising avenue for enhancing protection against economically significant viral diseases in chickens.
