PATHOGENETIC SIGNIFICANCE OF PROTEIN KINASE JNK-1 INDUCED BY ULTRAVIOLET IN PERIPHERAL BLOOD LYMPHOCYTES FROM SLE PATIENTS

对象和方法：采集10例SLE患者和5例健康志愿者的外周血淋巴细胞(PBL)。将5毫升LX10^6/mlPBL转移到培养皿中，并以紫外线照射20s(波长302 nm，8 mW/cm2)的未照射PBL作为对照。(1)培养7天后，用双抗体夹心法测定血清中的免疫球蛋白G、IL-2、IL-6水平，用载脂蛋白2.7或抗Annexin 5单抗检测细胞凋亡率。(3)用NP-40裂解PBL后，用Western印迹技术检测JNK-1的丝氨酸磷酸化水平。结果：(1)紫外线照射后，SLE患者外周血中Ig G、IL-2和IL-6的产生明显增加，而NHV中约75%的PBL呈apo 2.7或Annexin 5阳性，而SLE患者中95%的PBL呈凋亡状态。推测SLE-PBL免疫抑制功能亚群对紫外线照射等细胞凋亡刺激敏感，照射后SLE-PBL作为一个整体被激活，产生细胞因子和免疫球蛋白G。JNK-1丝氨酸磷酸化可能参与了紫外线刺激对PBL的激活作用。SLE-PBL免疫抑制功能亚群可能存在缺陷。

Objects and Methods : Peripheral blood lymphocytes (PBL) were obtained from lOSLE patients and 5 normal healthy volunteers (NHV). Five mililiter of lx 10^6/ml PBL was transfered to the Petri dish and UV-irradiated for 20 seconds (wave length 302 nm, 8 mW/cm^2) Non-irradiated PBL were used as the control. (1) The levels of IgG, IL-2 & IL-6 were measured by ELISA after 7 day-culture of PBL.(2) The ratio of apoptotic cells was determinied by using the monoclonal antibodies (apo.2.7 or anti- annexin 5 antibody). (3) The serine-phosphorylation of JNK-1 was investigated by Western blot technique after PBL were lysed with NP-40.Results : (1) The production of IgG, IL-2 and IL-6 was increased after UV-irradiation in SLE, but not in NHV.(2) After UV irradiation, about 75% of PBL were positive for apo 2.7 or annexin 5 in NHV whereas 95% of PBL were apoptotic in SLE.(3) The serine-phosphorylation of JNK-1 was more apparent after UV irradiation in SLE than in NHV.Discussion : According to my data, there can be a speculation that the immunosuppressive functional subpopulation of SLE-PBL was senseitive to apoptotic stimuli such as UV-irradiation and then SLE-PBL, as a whole, was activated to produce cytokines and IgG after irradiation. And the serine-phosphorylation of JNK-1 could be involved in PBL activation via UV stimulation. There might be a defect in immunosuppressive functional subpopulation of SLE-PBL.