Application of an Endothelial Cell Culture Assay for the Detection of Neutralizing Anti-Clostridium Perfringens Beta-Toxin Antibodies in a Porcine Vaccination Trial

Toxins (Basel). 2019 Apr 15;11(4):225. doi: 10.3390/toxins11040225.


Beta-toxin (CPB) is the major virulence factor of Clostridium perfringens type C, causing hemorrhagic enteritis in newborn pigs but also other animals and humans. Vaccines containing inactivated CPB are known to induce protective antibody titers in sow colostrum and neutralization of the CPB activity is thought to be essential for protective immunity in newborn piglets. However, no method is available to quantify the neutralizing effect of vaccine-induced antibody titers in pigs. (2) Methods: We developed a novel assay for the quantification of neutralizing anti-CPB antibodies. Sera and colostrum of sows immunized with a commercial C. perfringens type A and C vaccine was used to determine neutralizing effects on CPB induced cytotoxicity in endothelial cells. Antibody titers of sows and their piglets were determined and compared to results obtained by an ELISA. (3) Results: Vaccinated sows developed neutralizing antibodies against CPB in serum and colostrum. Multiparous sows developed higher serum and colostrum antibody titers after booster vaccinations than uniparous sows. The antibody titers of sows and those of their piglets correlated highly. Piglets from vaccinated sows were protected against intraperitoneal challenge with C. perfringens type C supernatant. (4) Conclusions: The test based on primary porcine endothelial cells quantifies neutralizing antibody activity in serum and colostrum of vaccinated sows and could be used to reduce and refine animal experimentation during vaccine development.

Keywords: C. perfringens type C; beta-toxin; cell culture assay; necrotizing enteritis; neutralizing antibodies; primary porcine endothelial cell.

Publication types

  • Randomized Controlled Trial, Veterinary
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Neutralizing / blood
  • Antibodies, Neutralizing / immunology*
  • Bacterial Toxins / genetics
  • Bacterial Toxins / immunology*
  • Bacterial Vaccines / administration & dosage*
  • Biological Assay
  • Cells, Cultured
  • Colostrum / immunology*
  • Endothelial Cells / drug effects
  • Endothelial Cells / immunology
  • Female
  • Recombinant Proteins / pharmacology
  • Swine
  • Vaccination


  • Antibodies, Neutralizing
  • Bacterial Toxins
  • Bacterial Vaccines
  • CPB protein, Clostridium perfringens
  • Recombinant Proteins