Circulating Endothelial Cells And S100A12 In Kawasaki Disease

【Objective】Kawasaki disease (KD) as known as mucocutaneous lymphode syndrome (MCLS) is an acute vasculitis syndrome primarily affecting small and medium-sized arteries, especially coronary artery. Even though intravenous immunoglobulin (IVIG) can shorten the course of KD and reduce the possibility of coronary artery lesions (CAL), there are about 10% of KD patients having no response to IVIG, who may have high incidence of CAL. In many countries, KD has become the commonest acquired heart disease among children and important reason for CAL in adults. The pathogenesis of CAL is still unknown and there is no laboratory index helping to know IVIG-non-responder (IVIG-NR) and CAL in early time.Recently, many researchers had considered circulating endothelial cells (CECs) and S100A12 as important evidences of CAL and vasculitis. In this study, a combination of monoclonal antibody and flow cytometry (FCM) was applied to evaluate the ratio of CECs to mononuclear cells (CECs/MNC), the positive rate of S100A12 on CECs (CECs-S100A12/CECs) and the fluorescence intensity of S100A12 (FI-S100A12-CECs) in order to investigate the probable pathogenesis of CAL and provide clinical evidence for simple way of predicting IVIG-NR-KD and CAL in early time.【Material and methods】55 KD patients in acute stage (A-KD): 29 male patients and 26 female patients were included with an average age of 34.00±25.80 months, varying from 3 to 128 months, all of whom were received IVIG (1g/kg/d for 2 days onset within 10 days) and aspirin. 5 cases had IVIG-NR and 9 cases complicated with CAL.29 KD patients in subacute stage (SA-KD): 18 male patients and 11 female patients were included with an average age of 35.55±31.99 months varying from 6 to 128 months, of whom 6 patients had IVIG-NR and 9 cases complicated with CAL.14 KD patients in convalescent stage (C-KD): 8 male patients and 6 female patients were included with an average age of 33.29±23.62 months varying from 7 to 72 months, of whom 5 cases had IVIG-NR and 6 cases complicated with CAL.26 healthy control cases (control): 13 male cases and 13 female cases were included, with an average age of 23.92±19.87 months varying from 4 to 85 months.FCM and monoclonal antibody were applied to evaluate the CECs/MNC, CECs-S100A12/CECs and FI-S100A12-CECs in both of KD patients and healthy control cases. 【Results】1.Compared with healthy children, the CECs/MNC, CECs-S100A12/CECs and FI-S100A12-CECs rose obviously among A-KD patients (P<0.05). In those patients, the CECs/MNC, CECs-S100A12/CECs and FI-S100A12-CECs were 0.39(0.04-11.10)%, (67.58±24.45)% and 214.00(9.99-2511.80) respectively, while in healthy controls, the data were 0.24(0.05-1.10)%, (39.35±19.58)% and 23.26(12.07-237.30) respectively.The CECs/MNC, CECs-S100A12/CECs and FI-S100A12-CECs were 0.16(0.01-1.00)%, (65.99±22.40)% and 246.00(17.40-2613.00) respectively in SA-KD patients after using IVIG Compared with A-KD before IVIG, the CECs/MNC in SA-KD decreased significantly (P=0.001), even to 0.24(0.06-1.70)% in C-KD patients, but the CECs-S100A12/CECs and FI-S100A12-CECs kept on high level.2. The CECs/MNC in A-KD patients with IVIG-NR was 0.38(0.13-6.25)% while the ratio was 0.40(0.04-11.48)% in patients with IVIG responder (P>0.05). The CECs-S100A12/CECsc and FI-S100A12-CECs were (69.34±26.81)%, 110.40(23.39-1622.00) in A-KD patients with IVIG-NR, while the data were (67.41±24.49)%, 228.15(9.88-2669.95) in A-KD patients with IVIG responder (P>0.05). The CECs/MNC was 0.15(0.01-1.00)% in SA-KD patients with IVIG responder, presenting obvious decrease compared with A-KD (P<0.01). The CECs/MNC was 0.29(0.11-0.65)% in SA-KD patients with IVIG-NR, presenting no obvious change compared with A-KD.3. The CECs/MNC was 0.36(0.07-0.70)% in A-KD patients with CAL, while ratio was 0.50(0.03-11.79)% in patients without CAL (P>0.05). The CECs-S100A12/ CECsc and FI-S100A12-CECs were (62.50±26.37)%, 119.00(9.25-3105.00) in A-KD patients with CAL, while the data were (68.58±24.24)%, 264.50(10.76-1604.68) in patients without CAL (P>0.05). The CECs/MNC was 0.15(0.01-1.00)% in SA-KD patients with normal coronary artery, presenting obvious decrease compared with acute stage (P<0.01). The CECs/MNC was 0.21(0.08-0.83)% in SA-KD patients with CAL, presenting no obvious decrease compared with acute stage.【Conclusions】1. The CECs/MNC rises in A-KD patients, manifesting vessel endothelium being attacked by inflammatory factor, CECs increasing. The high ratio of CECs/MNC might indicate high possibility of CAL and IVIG-NR in SA-KD patients.2. Compared with A-KD patients, the ratio of CECs/MNC dropped after using IVIG in SA-KD and C-KD, indicating IVIG may restrain the attack of vessel endothelium by inflammatory factor, decrease the damage of endothelia cell and prevent coronary artery lesion.3. The CECs-S100A12/CECsc and FI-S100A12-CECs can be an assistant index to diagnose KD.4. It indicates CAL that CECs/MNC increase in C-KD again.