B淋巴细胞的活化标志及其在慢性肾衰竭的临床意义

Activation marker for B lymphocyte and its clinical significance in chronic renal failure

目的 以CD2 3分子作为B淋巴细胞活化的标志 ,观察慢性肾衰 (CRF)维持性血液透析 (HD)患者B淋巴细胞激活状态。方法 以金黄色葡萄球菌CowanI株菌体 (SAC)与健康人外周血单个核细胞 (PBMC)数 5 0∶1的比例 ,以终浓度 5 0 μg/L的佛波醇酯 (PMA)分别培养PBMC 12h、2 4h和 5 0h ;用生物素链霉亲和素免疫细胞化学法 (BSA ICC)检测CD2 3 +细胞百分率 ;以速率散射比浊法检测SAC或PMA刺激 5 0h的上清液中IgG水平 ;以双抗体夹心ELASA检测血清可溶性CD2 3 (sCD2 3 )。结果 未经刺激的健康人和CRF HD患者PBMC中CD2 3 +细胞百分率分别为 ( 6.6± 0 .9) %和 ( 17.9± 3 .7) %差异有显著性 (P <0 .0 1)。健康人PBMC经SAC刺激12h、2 4h、5 0h后CD2 3 +细胞百分率分别为 ( 8.8± 1.7) %、( 13 .0± 1.6) %和 ( 16.2± 1.9) % ,与无药对照管比较差异有显著性 (P <0 .0 1) ;经PMA刺激 12h、2 4h后CD2 3 +细胞百分率分别为 ( 17.0± 2 .3 ) %、( 3 1.5± 2 .9) % ,与无药对照管比较差异有非常显著性 (P <0 .0 0 1) ;经SAC或PMA刺激 5 0h培养上清液IgG含量分别为 ( 9.1± 4.6) g/L和 ( 14.6± 5 .7)g/L ,后者与无药对照管 ( 7.3± 3 .7g/L)差异有显著性 (P <0 .0 1)。健康人和CRF HD患者血清sCD2 3含量分别为 ( 1.8± 0 .

Objective To observe the preactivation status of B cells from chronic renal failure hemodialysis (CRF-HD) patients, with CD23 molecule as an activation marker of B lymphocyte. Methods The peripheral blood mononuclear cells (PBMCs) from healthy volunteers (HVs) were stimulated with SAC (SAC to PBMC was in the ratio of 50 to 1) or PMA (final concentration 50 μg/L) and were harvested in 12 h, 24 h and 50 h respectively. The percentage of CD23 positive cells in PBMCs were detected with biotin-streptavidin immunocytochemistry (BSA-ICC) method, The IgG in supernantant of cultured PBMCs stimulated with SAC or PMA for 50 h was measured with rate immuno-nephelometry method. The soluble CD23 (sCD23) in serum from patients and heathy volunteers was determined with double antibody sandwich ELISA. Results The percentages of CD23 positive cells in unstimulated PBMCs from HVs and CRF-HD patients were 6.6±0.9% and 17.9±3.7% respectively, there was significat difference (P<0.01). The percentages of CD23 positive cells in PBMCs from HVs stimulated with SAC for 12 h, 24 h and 50 h were 8± 1.7%, 13.0±1.6% and 16.2±1.9% respectively, there was significant difference, compared with unstimulated control (P<0.01). The pecentages of CD23 positive cells in PBMCs from HVs stimulated with PMA for 12 h and 24 h were 17.0±2.3% and 31.5±2.9%, there was extremely significant difference, compared with unstimulated control (P<0.001). The IgG concentrations in supernatant of cultured PBMCs stimulated with SAC or PMA for 50 h were 9.1 g/L and 14.6±5.7 g/L respectively, the later was compared with unstimulated control (7.3±3.7 g/L), there was significat difference (P<0.01). The concentrations of sCD23 in serum from HVs and CRF-HD patients were 1.8±0.1 U/ml and 13.7±5.0 U/ml respectively, there was extremely significant difference (P<0.001). Conclusions SAC or PMA are possessed of the strog function activating B lymphocytes and promoting the expression of CD23 molecule and IgG synthesis. There is preactivation status of B lymphocytes of CRF-HD patients, it is one of causes leading to immune function disorder of CRF-HD patients.