ABSTRACT
An international collaborative study (coded BSP083) was performed under the aegis of the Biological Standardisation Programme supported by the Council of Europe and the European Commission, with the aim of replacing the in vivo challenge assays for potency determination of combined acellular pertussis (aP) vaccines by a refined procedure also allowing reduction of animal use. This study investigates whether the immunogenicity of aP vaccine components could be assayed in a guinea pig (gp) serology model, using the same vaccine immunising doses as for D and T components potency testing, instead of using separate animals as is currently done. The BSP83 project is a follow up of 3 former collaborative studies (coded BSP019, BSP034 and BSP035) on serological methods for the potency testing of tetanus (T) and diphtheria (D) vaccines for human use. The use of gp instead of mice serology has the advantage of providing a larger volume of good quality antiserum for the assay of several vaccine components in the same sample, hence providing the opportunity for animal sparing. The results of Phase I of the study demonstrated that gp serology may be a useful method for the immunogenicity assay of acellular pertussis vaccines. This was confirmed in Phase II of the study, using 7 different combined aP vaccines in an international collaborative study involving 17 laboratories from both public and private sectors. Clear dose-response relationships were observed for different vaccines by ELISA, for antibodies against aP antigens, i.e. pertussis toxin (PT), filamentous haemagglutinin (FHA), fimbrial agglutinogens-2/3 (Fim 2/3) and pertactin (PRN). Intra- and inter-laboratory variations of aP ELISA results were found to be within an acceptable range. For some combined vaccines, however, the range of vaccine dilutions for immunisation confirmed to be optimal for D and T potency testing may not provide optimal dose-response for all aP components. Method adjustments may thus be required and suitability should therefore be demonstrated for each vaccine combination and product prior to the application of this assay. The results of this study support the use of the gp serological method for the determination of the immunogenicity of aP vaccines. The application of the method for batch release testing of combined D, T and aP vaccines could significantly contribute to the implementation of the 3R principles through reduction of animal use and improved animal welfare, whilst reducing costs. As an outcome of this study, the Group of Experts No. 15 on Sera and Vaccines of the European Pharmacopoeia (Ph. Eur.) decided in February 2009 to include the gp serological assay as an example in the Ph. Eur. General chapter 2.7.16. on acellular pertussis vaccine assay.
