摘要
目的探讨核心岩藻糖基转移酶(Fut8)对B细胞抗体产生的影响。方法小鼠经OVA免疫后,取其脾脏分离单个淋巴细胞,应用流式细胞术分析小鼠脾脏细胞中B细胞群的分布情况;ELISPOT实验检测小鼠B细胞产生抗体的能力;收集小鼠OVA免疫过程中第0、14、28天的血清,ELISA技术检测小鼠血清中抗体的效价。结果 Fut8基因缺失导致小鼠脾脏细胞中所有的B细胞和成熟B细胞的数量下降(P<0.05);Fut8^(-/-)小鼠和Fut8^(+/+)小鼠抗体产生细胞的平均数量分别为16个和27个,与Fut8^(+/+)小鼠相比,Fut8^(-/-)小鼠B细胞产生抗体的能力下降(P<0.05);第1次免疫之后Fut8^(-/-)和Fut8^(+/+)小鼠抗-OVA IgG抗体的效价分别为1∶12 800和1∶51 200,第2次免疫之后,抗体的效价分别为1∶102 400和1∶409 600,Fut8的缺失导致血清中抗体效价下降(P<0.05)。结论核心岩藻糖基化对B细胞的抗体产生具有重要的调节作用。
Objective To explore the role of core fucosyltransferase(Fut8)on Ab production.MethodsSPL lymphocytes were isolated from OVA-immunized mice.Population of B cells was analyzed by flow cytometry.The frequency of Ab secreting cells was detected by enzyme-linked immunospot assay(ELISPOT).Fut8^+/+and Fut8^-/-mice were immunized with OVA,and the immune sera were collected at 0,14 and28 d.The Ab titers were detected by enzyme linked immunosorbent assay(ELISA).Results We found that lack of Fut8 gene impaired B cell generation in the spleen after OVA immunization.The number of IgG-producing cells of Fut8^-/-and Fut8^+/+ SPLs was 16 and 27,respectively.Compared with that of Fut8^+/+ SPLs,the frequency of IgG-producing cells was significantly reduced in Fut8^-/- SPLs(P〈0.05).ELISA analysis showed that the titers of anti-OVA IgG in the immune sera of Fut8^-/- and Fut8^+/+ mice were 1∶12 800 and1∶51 200,respectively;after the first immunization(14 d)and the boost immunization(28 d),they reached 1∶102 400 and 1∶409 600,respectively.These results suggested that the disruption of Fut8 led to reduction in Ab production(P〈0.05).Conclusion These results show that core fucosylation modified by Fut8 plays an important role in the regulation of Ab production.
出处
《西安交通大学学报(医学版)》
CAS
CSCD
北大核心
2016年第5期627-630,共4页
Journal of Xi’an Jiaotong University(Medical Sciences)
基金
国家自然科学基金资助项目(No.31270864
30972675)~~