应用流式细胞仪检测斑点叉尾鮰红细胞免疫粘附功能

制备荧光标记的致敏酵母及未致敏酵母,应用流式细胞仪检测斑点叉尾鮰全血体系与洗涤红细胞体系下红细胞C3b受体(erythrocyte C3breceptors,E-C3bR)与红细胞免疫复合物(erythrocyte immune complexes,E-IC)花环率,同时检测未灭活血清与灭活血清(58℃处理30min)对洗涤红细胞免疫粘附功能的调控作用.结果显示,斑点叉尾鮰红细胞能有效粘附致敏酵母及未致敏酵母,其灭活血清组E-C3bR与E-IC花环率(分别为16.78%和13.51%)明显高于未灭活血清组(分别为5.99%和6.99%)与生理盐水组(分别为8.59%和8.30%);未灭活血清组E-C3bR与E-IC花环率明显低于生理盐水组.试验结果表明斑点叉尾鮰红细胞具有免疫粘附功能,并且血清中存在促进红细胞免疫粘附功能的热稳定因子及抑制其免疫粘附功能的热不稳定因子.该试验不仅从技术上促进了红细胞免疫功能的研究,而且有助于全面客观地了解鱼类的红细胞免疫功能,为鱼类的病害监测及其防治奠定基础.

On Flow Cytometric Assessment of Channel Catfish (Ictalurus Punctatus)Erythrocytes in Immune Adherence Function

Using FITC-labeled yeasts and flow cytometry,the immune adherence rates of Channel Catfish(Ictalurus Punctatus) erythrocytes to non-sensitized yeasts(E-IC) and sensitized yeasts(E-C3bR) have been investigated.The Channel Catfish erythrocytes are in whole blood conditions and in washed conditions,respectively.The washed Channel Catfish erythrocytes are dealt with autologous serum,autologous inactivated serum(58 ℃ 30 min)and 0.68 % physiological saline,respectively.Our experiment indicates that the rates of E-C3bR and E-IC of the washed erythrocytes which are dealt with autologous inactivated serum(58 ℃ 30 min)are significantly higher than those of the washed erythrocytes dealt with autologous serum and 0.68 % physiological saline.Meanwhile,the rates of E-C3bR and E-IC of the washed erythrocytes which are dealt with autologous serum are significantly lower than those of the control group.Further study shows that the rates of E-C3bR and E-IC in whole blood conditions are both nearly as the same as those of the washed erythrocytes dealt with autologous serum.Our results show that the erythrocytes of Channel Catfish could effectively adhere to non-sensitized yeasts and sensitized yeasts,which is regulated by the thermo-stable enhancer and thermo-labile inhibitor in sera.By the way,the rates of E-C3bR and E-IC detected by flow cytometry are in agreement with those detected by the optical microscope on the whole,which suggests that it is feasible to investigate the immune adherence function of erythrocytes with flow cytometry.This study could not only technically boost the study of the immune adherence of erythrocytes,but also provide an insight into the immune defense system in fishes and build a basis for the detection and prevention of fish diseases.