EB病毒膜蛋白gp350对培养CD5^+B细胞产生天然自身抗体的影响

Effects of Epstein-Barr virus glycoprotein-350 on cultured CD5^+B cells in producing natural autoantibody

目的 :探讨EB病毒膜表面糖蛋白gp35 0是否刺激培养的CD5 + B细胞产生具有天然自身抗体性质的免疫球蛋白 .方法 :无菌采集新生儿脐带血 ,RosetteSepTM法分离总B细胞 ,免疫磁珠分离CD5 + B细胞 ,将分离得到的总B ,CD5 + B和CD5 -B 3组细胞分别用EB病毒、紫外线照射灭活的EB病毒(UV EBV)、gp35 0及TPA刺激 ,于培养的 1 0～ 30d间 ,每 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 .结果 :EB病毒感染的3组细胞于培养的第 7日发生转化 ,表现为细胞成团生长 ,体积增大 ,不断增殖 ,1 4d以后的培养上清中IgM ,AKautoAbIgMA值较对照组显著升高 (P <0 .0 1 ) ;紫外线照射灭活的EB病毒、gp35 0刺激的细胞存活 1 5d ,1 4dCD5 + B细胞培养上清中IgMA值高于对照组 (P <0 .0 5 ) ;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 .结论 :EB病毒可能通过其膜表面糖蛋白gp35 0刺激培养的CD5 +

AIM: To explore the effects of Epstein Barr virus glycoprotein 350 on cultured CD5 +B cells in producing natural autoantibody. METHODS: Blood of umbilical cord was collected under sterile condition and B cells were isolated by the method of RosetteSep TM . CD5 +B cells and CD5 -B cells were obtained by mouse IgG magnetic beads. Three groups of B cells, CD5 +B cells and CD5 -B cells were stimulated by Epstein Barr virus, ultraviolet radiation inactivated Epstein Barr virus, gp350 and TPA respectively. ELISA was used to detect immunoglobulin G, immunoglobulin M and antikeratin autoantibody in the supernatant at the time of 10, 14, 18, 22, 26 and 30 days after stimulation. Healthy adult blood serum was used as positive control and unstimulated cells as negative control. RESULTS: The three groups of cells stimulated by Epstein Barr virus were all transformed. The cells became larger, grew in mass and kept on proliferating. The A value of IgM, AK auto Ab IgM in the supernatant since the 14th day increased significantly compared with that of the control ( P <0.01). The cells stimulated by ultraviolet radiation inactivated Epstein Barr virus and gp350 survived 15 days and on the 14th day, the A value of IgM in the supernatant of CD5 +B cells increased compared with that of the control ( P <0.05). The TPA stimulated and unstimulated cells survived 7 days, and no immunoglobulins was observed. CONCLUSION: Epstein Barr virus can stimulate cultured CD5 +B cells by its glycoprotein 350 in producing immunoglobulin with the characteristics of natural autoantibody.