Turner S, Bakker N P, t Hart B A, Holt P J, Morgan K
Department of Rheumatology, University of Manchester Medical School, UK.
Clin Exp Immunol. 1994 May;96(2):275-80. doi: 10.1111/j.1365-2249.1994.tb06553.x.
Sera from eight rhesus monkeys that had been immunized with native bovine type II collagen were tested for antibodies to cyanogen bromide peptides (CB peptides) of type II collagen by Western blotting. The monkeys produced IgG antibodies to a number of different CB peptides, with five out of eight animals producing antibodies to the CB-11 peptide (four arthritic, one non-arthritic). Antibody epitopes on the CB-11 peptide of bovine type II collagen recognized by these sera were investigated by epitope mapping. Peptides (8-mers overlapping by seven amino acids) representing the CB-11 region were synthesised and the sera screened for binding to these peptides to determine areas of high IgG antibody binding to this region of type II collagen. The profiles obtained were not identical, though there were some epitopes that were commonly recognized. Antibodies to one epitope, also present in human type II collagen, were found only in the sera of two animals with the severest arthritis. The technique of epitope mapping has successfully identified a number of epitopes within the CB-11 peptide of type II collagen recognized by antibodies from bovine type II collagen-immunized monkeys. Studies on the relevance of responses to the identified epitopes can now be undertaken.
通过蛋白质印迹法检测了八只经天然牛II型胶原蛋白免疫的恒河猴血清中针对II型胶原蛋白溴化氰肽(CB肽)的抗体。这些猴子产生了针对多种不同CB肽的IgG抗体,八只动物中有五只产生了针对CB - 11肽的抗体(四只患关节炎,一只未患关节炎)。通过表位作图研究了这些血清所识别的牛II型胶原蛋白CB - 11肽上的抗体表位。合成了代表CB - 11区域的肽(八肽,氨基酸重叠七个),并检测血清与这些肽的结合情况,以确定IgG抗体与II型胶原蛋白该区域高结合的区域。尽管存在一些共同识别的表位,但所获得的图谱并不相同。仅在两只患最严重关节炎动物的血清中发现了针对一种表位的抗体,该表位也存在于人类II型胶原蛋白中。表位作图技术已成功鉴定出II型胶原蛋白CB - 11肽内被牛II型胶原蛋白免疫猴子的抗体所识别的多个表位。现在可以对针对已鉴定表位的反应相关性进行研究。