EB病毒对脐带血细胞产生免疫球蛋白的影响

Effect of Epstein Barr (EB) Virus on Cultured Umbilical Cord B and CD5^+B Cells in Production of Immunoglobin

探讨EB病毒是否使培养的脐带血B细胞、CD5+B细胞产生免疫球蛋白及具有天然自身抗体性质的免疫球蛋白。无菌采集新生儿脐带血,RosetteSepTM法分离全B细胞,免疫磁珠分离CD5+B细胞,将分离得到的全B,CD5+B,CD5-B三组细胞分别用EB病毒、紫外线照射灭活的EB病毒(UVEBV)、TPA刺激,于培养的第10～30d,每间隔4d分别留取上清,ELISA法检测培养上清中的IgG,IgM及抗角蛋白自身抗体(AKautoAb)IgG,IgM,阳性对照为健康成人血清,阴性对照为未加刺激的培养细胞上清。EB病毒感染的3组细胞于培养的第7d发生转化,14d以后的培养上清中IgM,AKautoAbIgMOD值较对照组显著升高(p<0.01);UVEBV刺激细胞存活15d,其第14d的CD5+B细胞培养上清中IgMOD值高于对照组(p<0.05);TPA及未刺激的细胞存活7d,培养上清中未检测到免疫球蛋白。可见,EB病毒能使培养的脐带血B细胞、CD5+B细胞产生可能具有天然自身抗体性质的免疫球蛋白。

In order to investigate whether immunoglobin produced by cultured umbilical cord B cells and CD5++B cells possesses immunoglobin of natural autoantibody feature, neonatal umbilical cord blood was collected in sterile conditions and B cells were isolated with RosetteSep+{TM} method to obtain CD5++B cells with mouse IgG magnetic beads. And the CD5++B cells were divided into three groups and stimulated with EB virus, inactivated EB virus with ultraviolet radiation, and TPA respectively. During the tenth to thirtieth days in culture, supernatant was taken in every four days to test IgG, IgM, and antikeratin autoantibody (AK auto Ab) in the supernatant.The positive control is serum of healthy adult, and the negative control is the supernatant of cultured cells without any stimulation. The results showed that the three group cells stimulated by EB virus infection were all transformed and happened at the seventh day of cultivation, and became larger, and grew in mass and proliferation unceasing. The OD value of supernatant IgM, AK auto Ab, and IgM after 14 days' cultivation were increased significantly as compared with the control (p<0.01); and the cells stimulated by ultraviolet radiation inactivated EB virus survived for 15 days. The IgM OD value of CD5++B cells cultural supernatant was higher than control (p<0.05). TPA and unstimulated cells survived for 7 days, and no immunoglobin was tested in the cultural supernatant. It is thus clear that EB virus could stimulate cultured umbilical cord B and CD5++B cells to produce immunoglobin that may possess characteristics of natural autoantibody.;