Winsnes R, Sesardic D, Daas A, Terao E, Behr-Gross M-E
Norwegian Medicines Agency (NoMA), N-0950 Oslo, Norway.
Pharmeur Bio Sci Notes. 2009 Oct;2009(1):27-40.
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.
在欧洲委员会和欧盟委员会支持的生物标准化计划的支持下，开展了一项国际合作研究（编号为BSP083），目的是用一种改进的方法取代用于测定联合无细胞百日咳（aP）疫苗效力的体内攻毒试验，同时减少动物使用量。本研究调查了是否可以在豚鼠血清学模型中检测aP疫苗成分的免疫原性，使用与D和T成分效力测试相同的疫苗免疫剂量，而不是像目前这样使用单独的动物。BSP83项目是之前3项关于人用破伤风（T）和白喉（D）疫苗效力测试血清学方法的合作研究（编号为BSP019、BSP034和BSP035）的后续研究。使用豚鼠血清学而非小鼠血清学的优势在于，可为同一样本中几种疫苗成分的检测提供更大量的高质量抗血清，从而提供减少动物使用的机会。该研究第一阶段的结果表明，豚鼠血清学可能是检测无细胞百日咳疫苗免疫原性的一种有用方法。在该研究的第二阶段得到了证实，在一项涉及17个公共和私营部门实验室的国际合作研究中使用了7种不同的联合aP疫苗。通过酶联免疫吸附测定（ELISA）观察到不同疫苗针对aP抗原（即百日咳毒素（PT）、丝状血凝素（FHA）、菌毛凝集原-2/3（Fim 2/3）和百日咳杆菌黏附素（PRN））的抗体有明确的剂量反应关系。发现aP ELISA结果的实验室内和实验室间变异在可接受范围内。然而，对于一些联合疫苗，经证实对D和T效力测试最佳的免疫疫苗稀释范围可能无法为所有aP成分提供最佳剂量反应。因此可能需要调整方法，因此在应用该检测方法之前，应针对每种疫苗组合和产品证明其适用性。本研究结果支持使用豚鼠血清学方法测定aP疫苗的免疫原性。将该方法应用于联合D、T和aP疫苗的批签发检测，可通过减少动物使用量和改善动物福利，同时降低成本来显著促进3R原则的实施。作为本研究的成果，欧洲药典血清和疫苗专家第15组于2009年2月决定将豚鼠血清学检测作为欧洲药典通则2.7.16“无细胞百日咳疫苗检测”中的一个示例纳入。
