Development of a cell culture assay for the quantitative determination of vaccination-induced antibodies in rabbit sera against Clostridium perfringens epsilon toxin and Clostridium novyi alpha toxin.
Cell culture assays are possible alternatives to replace in vivo neutralization tests currently required for potency testing of clostridial vaccines. Cell culture assays based on the MDCK cell line and the Vero cell line which are sensitive to the Clostridium (C.) perfringens type D epsilon toxin and Clostridium novyi type B alpha toxin, respectively, were developed, and the test conditions were standardized. The antibody titres of vaccinated rabbits measured in vitro were compared with the results of current test procedures recommended by European Pharmacopoeia. The correlation coefficients calculated were significant for all sera tested. The cell culture assays proved to be sensitive, specific, reproducible and reliable. Therefore, these cell culture assays could be suitable in vitro alternatives to the in vivo mouse neutralization experiments required for potency tests of clostridial vaccines, but further validation studies are necessary.
Borrmann
E
E
Institute of Molecular Pathogenesis at the Federal Research Institute for Animal Health, Friedrich-Loeffler-Institut, Naumburger Str. 96 a, D-07743 Jena, Germany. erika.borrmann@fli.bund.de
Schulze
F
F
Cussler
K
K
Hänel
I
I
Diller
R
R
D000826:Animal Testing Alternatives / Q000379:methods* / Q000592:standards; D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood*; D001427:Bacterial Toxins / Q000276:immunology*; D001428:Bacterial Vaccines / Q000592:standards*; D002460:Cell Line; D002522:Chlorocebus aethiops; D003013:Clostridium / Q000276:immunology*; D004285:Dogs; D007106:Immune Sera / Q000276:immunology; D007118:Immunoassay / Q000379:methods; D007668:Kidney / Q000166:cytology; D011817:Rabbits; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013223:Statistics as Topic; D014709:Vero Cells
D000907:Antibodies, Bacterial; D001427:Bacterial Toxins; D001428:Bacterial Vaccines; C075216:Clostridium novyi alpha-toxin; C039811:Clostridium perfringens epsilon-toxin; D007106:Immune Sera