Luan Jun-Jun, Xing Guang-Qun
Renal Division, Affiliated Hospital of Qingdao University, No. 1677 Wu Tai Shan Road, Qingdao, 266555, People's Republic of China.
J Nephrol. 2017 Feb;30(1):63-71. doi: 10.1007/s40620-016-0336-z. Epub 2016 Jul 30.
Anti-neutrophil cytoplasmic antibodies (ANCA)-associated vasculitides (AAV) are a group of systemic autoimmune disorders characterized by necrotizing inflammation of small- to medium-sized blood vessels. The pathogenesis of patients with AAV are still in investigation. In this study, we explored the involvement of LL-37 and nucleic acids in AAV.
15 patients with AAV diagnosed according to the Chapel Hill definition between October 2014 and July 2015 in the department of Nephrology of Huangdao, affiliated Hospital of Qingdao University were enrolled. 16 patients with chronic bronchitis (CB) were selected as disease control group. 15 cases of healthy people from Medical Healthy Center were as healthy control group. Peripheral blood mononuclear cells (PBMCs) were collected from these groups and stimulated by LL-37and (or) two types of CpG-ODN for 7 days. The IFN-α and ANCA in vitro were measured by ELISA. The serum IFN-α, LL-37 and ANCA were measured also.
The serum level of IFN-α in AAV group was much higher than that in CB group (692.13 ± 407.28 vs 397.07 ± 211.62 pg/ml, p = 0.019), and that in healthy control group (692.13 ± 407.28 vs 251.54 ± 190.46 pg/ml, p < 0.001). The serum level of LL-37 in AAV group was much higher than that in CB group (101.18 ± 66.59 vs 40.23 ± 13.51 ng/ml, p < 0.001, and that in healthy control group (101.18 ± 66.59 vs 27.80 ± 16.86 ng/ml, p < 0.001). Also the level of IFN-α showed a significant positive relationship with ANCA in AAV group both in serum and in supernatant of cultured PBMCs stimulated by LL-37 and (or) CpG-ODN (r = 0.783, p = 0.001; r = 0.575, p = 0.064; r = 0.649, p = 0.031; r = 0.806, p = 0.003). In patients with AAV, the supernatant levels of IFN-α in cultured PBMCs stimulated by LL-37 and (or) CpG-ODN were higher than that without stimulating factor (p < 0.05). The supernatant level of IFN-α in cultured PBMC stimulated by LL-37 alone was lower than that stimulated by CpGA alone (699.57 ± 476.26 vs 2342.63 ± 2025.11 pg/ml, p = 0.001). But the supernatant level of IFN-α in cultured PBMCs stimulated by LL-37 alone was higher than in that stimulated by CpGB alone (699.57 ± 476.26 vs 153.35 ± 78.08 pg/ml, p < 0.001). The supernatant level of IFN-α in cultured PBMCs stimulated by both LL-37 and CpG-ODN was higher than that stimulated by LL-37 or CpG-ODN alone (2550.57 ± 2217.41 vs 699.57 ± 476.26 pg/ml, p = 0.003; 2550.57 ± 2217.41 vs 153.35 ± 78.08 pg/ml, p = 0.001; 2660.95 ± 391.31 vs 699.57 ± 476.26 pg/ml, p < 0.001; 2660.95 ± 391.31 vs 153.35 ± 78.08 pg/ml, p < 0.001). Either it is stimulated by LL-37 or CpG-ODN or both, the supernatant level of IFN-α in cultured PBMCs in AAV patients was the highest, that in healthy controls was the lowest. Either stimulated by LL-37 or CPG-ODN, or both, the levels of ANCA production in vitro in AAV groups were statistically significantly higher than those in CB group and healthy control group.
There were higher serum levels of LL-37 and IFN-α in patients with AAV. IFN-α could reach a higher level stimulated by LL-37 and nucleic acids both of which are related to infection. Patients with AAV have ANCA-producing B lymphocytes in the circulation even in remission stage. Infections could induce the relapse of AAV.
抗中性粒细胞胞浆抗体（ANCA）相关血管炎（AAV）是一组以中小血管坏死性炎症为特征的全身性自身免疫性疾病。AAV患者的发病机制仍在研究中。在本研究中，我们探讨了LL-37和核酸在AAV中的作用。
选取2014年10月至2015年7月在青岛大学附属医院黄岛院区肾内科就诊、根据 Chapel Hill 定义确诊的15例AAV患者。选取16例慢性支气管炎（CB）患者作为疾病对照组。选取健康体检中心的15例健康人作为健康对照组。采集这些组的外周血单个核细胞（PBMC），用LL-37和（或）两种类型的CpG-ODN刺激7天。采用ELISA法检测体外IFN-α和ANCA。同时检测血清IFN-α、LL-37和ANCA。
AAV组血清IFN-α水平显著高于CB组（692.13±407.28 vs 397.07±211.62 pg/ml，p = 0.019），也高于健康对照组（692.13±407.28 vs 251.54±190.46 pg/ml，p < 0.001）。AAV组血清LL-37水平显著高于CB组（101.18±66.59 vs 40.23±13.51 ng/ml，p < 0.001），也高于健康对照组（101.18±66.59 vs 27.80±16.86 ng/ml，p < 0.001）。此外，在AAV组中，血清和经LL-37和（或）CpG-ODN刺激的培养PBMC上清液中的IFN-α水平与ANCA均呈显著正相关（r = 0.783，p = 0.001；r = 0.575，p = 0.064；r = 0.649，p = 0.031；r = 0.806，p = 0.003）。在AAV患者中，经LL-37和（或）CpG-ODN刺激的培养PBMC上清液中的IFN-α水平高于无刺激因子时（p < 0.05）。单独用LL-37刺激的培养PBMC上清液中的IFN-α水平低于单独用CpGA刺激时（699.57±476.26 vs 2342.63±2025.11 pg/ml，p = 0.001）。但单独用LL-37刺激的培养PBMC上清液中的IFN-α水平高于单独用CpGB刺激时（699.57±476.26 vs 153.35±78.08 pg/ml，p < 0.001）。同时用LL-37和CpG-ODN刺激的培养PBMC上清液中的IFN-α水平高于单独用LL-37或CpG-ODN刺激时（2550.57±2217.41 vs 699.57±476.26 pg/ml，p = 0.
