title: Immunization with recombinant bivalent chimera r-Cpae confers protection against alpha toxin and enterotoxin of Clostridium perfringens type A in murine model.
abstract: Clostridium perfringens type A, an anaerobic pathogen is the most potent cause of soft tissue infections like gas gangrene and enteric diseases like food poisoning and enteritis. The disease manifestations are mediated via two important exotoxins, viz. myonecrotic alpha toxin (αC) and enterotoxin (CPE). In the present study, we synthesized a bivalent chimeric protein r-Cpae comprising C-terminal binding regions of αC and CPE using structural vaccinology rationale and assessed its protective efficacy against both alpha toxin (αC) and enterotoxin (CPE) respectively, in murine model. Active immunization of mice with r-Cpae generated high circulating serum IgG (systemic), significantly increased intestinal mucosal s-IgA antibody titres and resulted in substantial protection to the immunized animals (100% and 75% survival) with reduced tissue morbidity when administered with 5×LD(100) doses of αC (intramuscular) and CPE (intra-gastric gavage) respectively. Mouse RBCs and Caco-2 cells incubated with a mixture of anti-r-Cpae antibodies and αC and CPE respectively, illustrated significantly higher protection against the respective toxins. Passive immunization of mice with a similar mixture resulted in 91-100% survival at the end of the 15 days observation period while mice immunized with a concoction of sham sera and respective toxins died within 2-3 days. This work demonstrates the efficacy of the rationally designed r-Cpae chimeric protein as a potential sub unit vaccine candidate against αC and CPE of C. perfringens type A toxemia.
authors: Division of Microbiology, Defence Food Research Laboratory, Siddartha Nagar, Mysore 570011, Karnataka, India.
authors: Das
authors: D
authors: Shreya
authors: Division of Microbiology, Defence Food Research Laboratory, Siddartha Nagar, Mysore 570011, Karnataka, India.
authors: Siva R
authors: SR
authors: Uppalapati
authors: Division of Microbiology, Defence Food Research Laboratory, Siddartha Nagar, Mysore 570011, Karnataka, India. Electronic address: joseph@dfrl.drdo.in.
authors: Joseph J
authors: JJ
authors: Kingston
authors: Division of Microbiology, Defence Food Research Laboratory, Siddartha Nagar, Mysore 570011, Karnataka, India.
authors: Murali H
authors: MH
authors: Sripathy
authors: Division of Microbiology, Defence Food Research Laboratory, Siddartha Nagar, Mysore 570011, Karnataka, India.
authors: Harsh V
authors: HV
authors: Batra
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood / Q000276:immunology; D001427:Bacterial Toxins / Q000235:genetics / Q000276:immunology*; D001428:Bacterial Vaccines / Q000276:immunology; D018938:Caco-2 Cells; D002135:Calcium-Binding Proteins / Q000235:genetics / Q000276:immunology*; D045744:Cell Line, Tumor; D003015:Clostridium Infections / Q000276:immunology* / Q000517:prevention & control; D003016:Clostridium perfringens / Q000235:genetics / Q000472:pathogenicity; D004195:Disease Models, Animal; D004768:Enterotoxins / Q000235:genetics / Q000276:immunology*; D005260:Female; D006801:Humans; D007114:Immunization; D007116:Immunization, Passive; D007070:Immunoglobulin A / Q000097:blood / Q000276:immunology; D007074:Immunoglobulin G / Q000097:blood / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins / Q000494:pharmacology*; D010738:Type C Phospholipases / Q000235:genetics / Q000276:immunology*; D022223:Vaccines, Subunit / Q000494:pharmacology*
chemical_list: D000907:Antibodies, Bacterial; D001427:Bacterial Toxins; D001428:Bacterial Vaccines; D002135:Calcium-Binding Proteins; D004768:Enterotoxins; D007070:Immunoglobulin A; D007074:Immunoglobulin G; D011993:Recombinant Fusion Proteins; D022223:Vaccines, Subunit; C035598:enterotoxin, Clostridium; D010738:Type C Phospholipases; C040649:alpha toxin, Clostridium perfringens
keywords: Active protection; Alpha toxin; Chimeric protein r-Cpae; Clostridium perfringens type A; Enterotoxin; Passive protection