紫外线灭活的EB病毒蛋白对抗角蛋白自身抗体产生的调节作用

目的研究紫外线(UV)灭活和热处理的EB病毒刺激培养的人脐带血B细胞产生抗角蛋白自身抗体(AKautoAb)的作用。方法用淋巴细胞分离液分离人脐带血单一核细胞,L-亮氨酸甲酯去除单核细胞、NK细胞和细胞毒性T细胞,2-氨乙基硫脲溴化物处理的绵羊红细胞去除T细胞,从而获得纯化B细胞。用UV灭活和热处理EBV处理培养的B细胞。流式细胞仪检测UV灭活EBV组细胞CD5、CD3、CD4和CD8的表达。ELISA法检测培养上清液中AKautoAbIgG和IgM的产生,并与EBV转化B细胞产生的AKautoAbIgG和IgM作对比。结果细胞培养14d时,未检测到T细胞,CD5 B细胞占43%;28d时,CD5 B细胞占47%。UV灭活EBV组AKautoAbIgG18d以后各时间点有显著变化(P<0.05),AKautoAbIgM26d与其他时间点差异有显著性(P<0.05)。热处理EBV组AKautoAbIgG和IgM无明显变化(P>0.05)。EBV转化B细胞40d,AKautoAbIgG和IgM的产生与同期的UV灭活EBV组差异有显著性(P<0.05)。结论UV灭活EBV有诱导AKautoAb产生的作用,而热处理EBV不能诱导抗体的产生,提示灭活EBV诱导AKautoAb产生可能是通过EBV的蛋白成分实现的。

UV-inactivated Epstein-Barr Virus Protein Regulating the Production of Antikeratin Autoanfibodies

Objective To investigate UV- or heat-inactivated Epstein-Barr virus(EBV)stimulating the production of anti-keratin autoantibody(AK auto Ab)in cultured human umbilical cord blood B cells. Methods Mononuclear cells were isolated routinely from umbilical cord blood, in which monocytes, NK cells and cytotoxicity T cells were eliminated by L-leucine methyl ester method, and T cells were removed by sheep red blood cells(SRBCs)treated with 2-amino ethyl-isothiouronium bromide (AET). The purified B cells were treated with UV- or heat-inactivated EBV respectively and then cultured in complete IMDM. CD5, CD3, CD4 and CD8 cells were detected by flow cytometry. IgG and IgM of AK auto Ab were measured by ELISA in the supernatant which came from the B cells treated by UV-inactivated EBV or EBV-transformed B cells respectively. Results In UV-inactivated EBV group CD5 B cells accounted for 43% and 47% of all cells detected on the 14th and 28th day, respectively. No CD3, CD4 and CD8 cells were detected during this period. In UV-inactivated EBV group the AK auto Ab of IgG and IgM increased significantly on the 18th and 26th day, respectively (P < 0.05, P < 0.05), but no significant difference in heat-inactivated EBV group (P > 0.05). On the 40th day the AK auto Ab of IgG and IgM were significantly higher in EBV-transformed B cell group than those in UV-inactivated EBV group. Conclusions UV-inactivated EBV is able to induce AK auto Ab production but heat-inactivated EBV does not, which suggests that EBV protein might be the effective agent in inducing the production of AK auto Ab.