戊二醛聚合猪血红蛋白免疫学特性分析

The analysis of immunological properties of glutaraldehyde-polymerized porcine hemoglobin

目的阐明加弗氏佐剂条件下,戊二醛聚合猪血红蛋白(glutaradehyde polymerized porcine he-moglobin,pPolyHb)对不同种类动物的免疫原性,初步探讨pPolyHb与人、牛血红蛋白的抗原决定簇的异同。方法在加弗氏佐剂的条件下,pPolyHb多次皮下免疫小鼠、大鼠和兔子,间接ELISA法检测抗血清IgG滴度;免疫印迹法分别检测小鼠、大鼠和兔子抗pPolyHb抗体与猪、牛、人、小鼠、大鼠、兔子、pPolyHb和戊二醛聚合牛血红蛋白之间的交叉反应。结果在加弗氏佐剂的条件下,pPolyHb能够刺激动物产生较高滴度的IgG,不同种类的动物产生IgG滴度不同;小鼠、大鼠和兔子抗pPolyHb抗体与不同来源的血红蛋白之间的交叉反应有明显的差异。结论在有弗氏佐剂的条件下,pPolyHb有较强的免疫原性。但是,对不同种类动物免疫刺激的强弱不同;对不同种类的动物,pPolyHb刺激抗体产生的抗原决定簇可能大部分相同,但也有部分不同,由此产生的抗体也不同;pPolyHb与人和牛血红蛋白均有一些相似的抗原决定簇,但也有一些相互独立的抗原决定簇。

Aim To demonstrate the immunogenicity of pPolyHb when it is injected into different species of animals,at the presence of frund′s adjuvant,and the differences of antigenic epitopes of pPolyHb,hHb and bHb.Methods pPolyHb was mixed with frund′s adjuvant and immunized mice,rats and rabbits by general methods respectively,antiserum were colleted and titers of anti-hemoglobin IgG in antisera were detected by indrect ELISA.The cross-reactions of IgG against pPolyHb from these animals with pPolyHb,bPolyHb,pHb,bHb,hHb,mHb,ratHb and rabHb were detected by Western blotting.Results At the presence of frund′s adjuvant,repeated immunization of pPolyHb resulted in increase titers and different IgG titers in different species of animals.Western blotting data showed there were significant differences among the cross-binding reactions of the specific IgG against pPolyHb from these three kinds of animals with pPolyHb,bPolyHb,pHb,bH,hHb,mHb,ratHb and rabHb.Conclusion At the presence of frund′s adjuvant,pPolyHb showed strong immunogenicity and elicited different IgG titers in different species of animals.It′s likely the antigenic determinants of pPolyHb were different or exhibit different immunogenicity when given to different species of animals,so the produced IgG in mice,rats or rabbites were not the same.Although pPolyHb,hHb and bHb share parts of antigen determinants,they have some different epitopes too.