Ibsen P H, Petersen J W, Heron I
Bacterial Vaccine Department, Statens Seruminstitut, Copenhagen, Denmark.
Vaccine. 1993;11(3):318-22. doi: 10.1016/0264-410x(93)90193-2.
The amounts of pertussis toxin (PT), filamentous haemagglutinin (FHA), 69 kDa outer membrane protein (69 kDa OMP) and agglutinogens (AGG) 2 and 3 in extracts from the Danish whole-cell pertussis vaccine were studied in quantitative capture ELISA. With the exception of PT, the most effective extraction of these antigens was by heating the bacteria at 60 degrees C for 30 min in 2 M urea followed by sonication for 45 s. Extraction by 1 M sodium chloride prior to sonication resulted in higher levels of antigenic and biologically active PT. On average, a single human dose of pertussis vaccine (approximately 16 opacity units) was found to contain 5520 ng FHA, 63 ng PT, 1061 ng 69 kDa OMP, 397 ng AGG 2, 534 ng AGG 3 and 4840 ng lipopolysaccharide (LPS). The antigen content of one dose of the Danish pertussis vaccine appears to be low compared with the amounts found in the acellular vaccines currently in use. These findings may have important implications for the evaluation of the protective substances and the immunogenicity of whole-cell as opposed to acellular pertussis vaccines.
采用定量捕获酶联免疫吸附测定法（ELISA），对丹麦全细胞百日咳疫苗提取物中的百日咳毒素（PT）、丝状血凝素（FHA）、69 kDa外膜蛋白（69 kDa OMP）以及凝集原（AGG）2和3的含量进行了研究。除PT外，这些抗原最有效的提取方法是将细菌在2 M尿素中于60℃加热30分钟，随后超声处理45秒。在超声处理前用1 M氯化钠进行提取，可得到更高水平的具有抗原性和生物活性的PT。平均而言，一剂人用百日咳疫苗（约16个浊度单位）含有5520 ng FHA、63 ng PT、1061 ng 69 kDa OMP、397 ng AGG 2、534 ng AGG 3和4840 ng脂多糖（LPS）。与目前使用的无细胞疫苗中发现的含量相比，一剂丹麦百日咳疫苗的抗原含量似乎较低。这些发现可能对评估全细胞百日咳疫苗与无细胞百日咳疫苗的保护物质及免疫原性具有重要意义。
