CAJ | 학술논문

采用C3b受体花环实验探究了牙鲆红细胞对不同种类抗原的免疫黏附能力,确定了影响免疫黏附作用的理化因子.实验选取创伤弧菌、金黄色葡萄球菌和啤酒酵母3种不同类别的抗原分别与牙鲆红细胞在优化条件下反应.结果表明,牙鲆红细胞对3种抗原均有免疫黏附作用,在20℃、0.1mol/L PBS~(++)缓冲液的反应条件下,牙鲆红细胞黏附创伤弧菌、金黄色葡萄球菌和啤酒酵母的C3b受体花环率分别为(19.00±1.01)%、(6.09±1.36)%和(3.42±0.00)%,对3种抗原的免疫黏附活性差异显著(P<0.05).利用AKTA快速蛋白液相层析系统的Superdex 200 GL型高效柱分离纯化牙鲆红细胞膜蛋白Ⅰ型补体受体(CR1),通过dot-ELISA法检测发现红细胞膜蛋白组分中含有CR1成分,表明牙鲆红细胞膜上表达了与高等脊椎动物有较高同源性的CR1分子.此实验首次证明了牙鲆红细胞具有发挥免疫功能的重要物质基础.
채용C3b수체화배실험탐구료아평홍세포대불동충류항원적면역점부능력,학정료영향면역점부작용적이화인자.실험선취창상호균、금황색포도구균화비주효모3충불동유별적항원분별여아평홍세포재우화조건하반응.결과표명,아평홍세포대3충항원균유면역점부작용,재20℃、0.1mol/L PBS~(++)완충액적반응조건하,아평홍세포점부창상호균、금황색포도구균화비주효모적C3b수체화배솔분별위(19.00±1.01)%、(6.09±1.36)%화(3.42±0.00)%,대3충항원적면역점부활성차이현저(P<0.05).이용AKTA쾌속단백액상층석계통적Superdex 200 GL형고효주분리순화아평홍세포막단백Ⅰ형보체수체(CR1),통과dot-ELISA법검측발현홍세포막단백조분중함유CR1성분,표명아평홍세포막상표체료여고등척추동물유교고동원성적CR1분자.차실험수차증명료아평홍세포구유발휘면역공능적중요물질기출.
The C3b receptor rosette technique was employed to investigate the erythrocytes immune adherence activity of left-eyed flounder (Paralichthys olivaceus) to different antigens under the optimized immune adherence conditions. Three different kinds of antigens (gram-negative bacteria, gram-positive bacteria and eukaryote) were chosen to incubate with erythrocytes in the 0.1 mol/L PBS~(++) buffer at 20℃ for 30 min. The results indicated that the flounder erythrocytes were able to adhere to all the three kinds of antigens with the C3b receptor rosette rates for Vibrio vulnificus, Staphylococcus aureus and Saccharomyces cerevisiae being (19.00±1.01)%, (6.09±1.36)% and (3.42 ± 0.00)% respectively. The external structures of the antigens were inferred to be responsible for the significant difference of rosette rates. Furthermore, erythrocyte membrane proteins were extracted and purified by the Superdex 200 FPLC gel chromatography to prepare the complement receptorⅠ (CR1) component. The Dot-ELISA assay confirmed there was the CR1 component in erythrocyte membrane proteins and the CR1 molecule homologous to higher vertebrate was expressed on erythrocytes of left-eyed flounder. Erythrocytes have been proved to possess key immune molecules and play an important role in the left-eyed flounder immune system in the study.