戊二醛聚合对猪、牛和人血红蛋白免疫学特性的影响

The Influence of Glutaraldehyde Polymerization on Immunological Properties of Porcine Hemoglobin,Bovine Hemoglobin and Human Hemoglobin

猪、牛、人血红蛋白或其相应的戊二醛聚合体分别经皮下或腹腔多次免疫小鼠,ELISA检测抗血清IgG滴度;免疫印迹法分别检测小鼠抗猪、牛、人血红蛋白或其相应的戊二醛聚合体IgG与猪、牛、人血红蛋白或其相应的戊二醛聚合体之间的交叉反应,探讨戊二醛聚合对不同种类血红蛋白免疫学特性的影响。结果表明:猪、牛、人血红蛋白的免疫原性均很弱,其相应的戊二醛聚合体免疫原性明显增强;猪、牛、人血红蛋白具有明显的抗原性差异;戊二醛聚合对这三类血红蛋白免疫学性质的影响完全不同,戊二醛聚合猪血红蛋白没有产生新的抗原决定簇,戊二醛聚合牛血红蛋白某些抗原决定簇被封闭,戊二醛聚合人血红蛋白形成了某些新的抗原决定簇。

To reveal the effects of glutaraldehyde polymerization on the immunological properties of porcine hemoglobin, bovine hemoglobin and human hemoglobin, repeated ip. or sc. administration of an immunizing dose of pHb, bHb, hHb or their glutaraldehyde cross-linked derivatives were given to mice to stimulate IgG responses, which was measured by indirect ELISA. Western blotting analysis of the antibody-antigen cross- binding reations between pHb, bHb, hHb or its related glutaraldehyde polymerized hemoglobin derivatives and anti-hemoglobin or anti-glutaraldehyde polymerized hemoglobin antibodies raised in mice were used in order to investigate the effects of glutaraldehyde polymerization on the immunological properties of the proteins concerned. The results indicated that hHb, bHb and pHb are all weakly immunogenic, while polymerization significantly increases the immunogenicity of all their glutaraldehyde-cross-linked derivatives. There exist clear species- specific antigen-determinants in hHb, bHb and pHb. The effects of glutaraldehyde polymerization on immunological properties of porcine hemoglobin, bovine hemoglobin and human hemoglobin are significantly different, there were no neo-antigens formed in pPolyHb, some of the determinants of bHb were covered because of glutaraldehyde polymerization, however, neo-antigens produced in hPolyHb. Therefore, it is necessary to study the immunological characteristics of different HBOCs products separately and should not inferred the characteristics of the products from different kinds of Hb even when they were made by the same method.