Development of Adjuvant-Free Bivalent Food Poisoning Vaccine by Augmenting the Antigenicity of Clostridium perfringens Enterotoxin

Abstract

Clostridium perfringens enterotoxin (CPE) is a common cause of food poisoning and hyperkalemia-associated death. Previously, we reported that fusion of pneumococcal surface protein A (PspA) to C-terminal fragment of CPE (C-CPE) efficiently bound mucosal epithelium so that PspA-specific immune responses could be provoked. In this study, we found that fusion of C-CPE with PspA augmented the antigenicity of C-CPE itself. These findings allowed us to hypothesize that fusion of C-CPE and another food poisoning vaccine act as a bivalent food poisoning vaccine. Therefore, we constructed an adjuvant-free bivalent vaccine against CPE and cholera toxin (CT), which is a major food poisoning in developing country, by genetically fusing CT B subunit to C-CPE. Because of the low antigenicity of C-CPE, immunization of mice with C-CPE alone did not induce C-CPE-specific immune responses. However, immunization with our vaccine induced both C-CPE- and CT-specific neutralizing antibody. The underlying mechanism of the augmented antigenicity of C-CPE included the activation of T cells by CTB. Moreover, neutralizing antibodies lasted for at least 48 weeks and the quality of the antibody was dependent on the binding activity of CTB–C-CPE to its receptors. These findings suggest that our fusion protein is a potential platform for the development of an adjuvant-free bivalent vaccine against CPE and CT.

Potential for Tight Junction Protein–Directed Drug Development Using Claudin Binders and Angubindin-1

“…TJ binders are classified as first-generation binders (toxins and their fragments), and second-generation binders (antibodies) [32,33,85]. The augmentation of the antigenicity of the first-generation binder, C-CPE, has been used to develop an adjuvant-free vaccine [97,99]. Second-generation binders are being used to develop cancer therapies (Table 2).…”

“…Nasal immunization with chimeric C-CPE-antigen did not induce mucosal injury [98]. The augmentation of the antigenicity of the first-generation binder C-CPE has been used to develop an adjuvant-free bivalent food poisoning vaccine [99]…”

Development, Structure, and Mechanism of Synthetic Antibodies that Target Claudin and Clostridium perfringens Enterotoxin Complexes

“…In humans CpE causes enterotoxemia; is the 3 rd most prevalent foodborne illness in the United States causing an estimated ~$400 million annual economic burden; and is the source of a further 4+ million food poisoning cases worldwide-some resulting in death (7,(9)(10)(11)(12). Unlike other human diseases caused by Clostridium perfringens toxins, CpE-associated ailments are not directly preventable nor treatable, and no vaccine exists against this food poisoning type (13). Because CpE is heat-resistant and produced by spore-formed Clostridium perfringens, cooking does little to reduce its pathogenicity (14).…”

Complete genome analysis and biological characterization of a novel Clostridium perfringens phage vB_CP_qdyz_P5

“…This bacterium is naturally present in soil, sewage, animals, and in the human intestinal tract [2]. It is the second most important foodborne pathogen in developed countries [3], with more than 4 million cases per year globally [4].However, due to the misuse of antibiotic, drug-resistant bacteria are becoming more…”