Submitted to: Microbial Pathogenesis
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 5/10/2025
Publication Date: 5/16/2025
Citation: Dhiraj, C., Shailes, B., N N 3. Madhusudan, T., Mostafa, G., Lillehoj, H.S., Sun, Z., Li, C.Z. 2025. Immunoinformatic-guided design and refinement of multi-epitope vaccines targeting Clostridium perfringens collagen adhesin protein (CNA) through CD4+ Th cells, CD8+ CTL, and B cell epitopes. Microbial Pathogenesis. 13:1147. https://doi.org/10.3390/ microorganisms13051147.
DOI: https://doi.org/10.3390/ microorganisms13051147

Interpretive Summary: Necrotic enteritis (NE) is a serious gut disease in poultry caused by an infection from a bacterium called Clostridium perfringens. This disease leads to major financial losses of estimated 6 billion US dollars in the global poultry industry. One reason for the increase in NE cases is the recent trend of reducing or removing antibiotics from animal feed, which had previously helped control such infections. As a result, developing effective vaccines against NE has become a top priority. The pathogenic bacterium Clostridium perfringens is a significant health threat to both humans and animals. One key factor in its ability to cause NE is a protein of 647 amino acids in length called collagen adhesion (CNA). Targeting CNA for vaccine development could be a promising way to protect poultry from this disease. In our approach, we developed a multi-pieces vaccine (MEV) designed to target CNA. This vaccine includes pieces of CNA that can stimulate the immune system, specifically for the cell-mediated system and antibody production. Using advanced computer tools, we identified and selected the best parts of CNA to include in the vaccine, ensuring it would be effective while minimizing allergic reactions. We created a vaccine containing 115 amino acids from CNA (about 1/5 of the native protein in length), representing the most important functional parts of the protein. This new vaccine shows strong potential in protecting poultry against Clostridium perfringens infections, offering similar immunity as vaccines based on the entire CNA protein.

Technical Abstract: Necrotic enteritis (NE) is a multifactorial enteric infectious disease caused by pathogenic Clostridium perfringens, leading to significant economic losses estimated at around USD 6 billion annually for the global poultry industry. The rising incidence of NE has been attributed to the voluntary reduction or elimination of antibiotic growth promoters in animal feed in recent years. Consequently, developing effective NE-specific vaccines has become a top priority for the poultry industry. C. perfringens poses a serious threat to both human and animal health, with collagen adhesion (CNA) serving as a critical virulence factor in NE pathogenesis among poultry. Targeting CNA as a prime target for vaccination offers a promising approach to reduce the impact of C. perfringens. Here, we have developed a comprehensive strategy for creating and refining multi-epitope vaccines against CNA, incorporating CD4+ helper T lymphocyte (Th), CD8+ cytotoxic T lymphocyte (CTL), and B-cell epitopes. Using bioinformatics tools and algorithms, we identified potential epitopes to enhance their immunogenicity and antigenicity while minimizing allergenicity. A 115-amino-acid peptide representing most of these epitopes has been created. This multi-epitope vaccine candidate demonstrated strong potential for providing immunity against pathogenic C. perfringens and NE infections in the terms of induction of IgG and IgM antibody levels, plasma cells, Th memory cells and CTL cells, comparable to those induced by the entire CNA protein.