抗α-1C微管蛋白抗体在系统性硬化症中的表达及临床意义

doi:10.19723/j.issn.1671-167X.2020.06.004

摘要/Abstract

摘要：

目的:检测抗α-1C微管蛋白(tubulin-α-1C)抗体在系统性硬化症(systemic sclerosis, SSc)患者血清中的表达,并探讨其潜在的临床意义。方法:入组SSc患者62例、系统性红斑狼疮(systemic lupus erythematosus, SLE)患者38例、原发性干燥综合征(primary Sj?gren’s syndrome, pSS)患者24例和健康对照组(health control, HC)30例,收集血清,采用酶联免疫吸附试验(enzyme linked immunosorbent assay, ELISA)分别检测各组血清中抗tubulin-α-1C抗体水平。同时,用标准实验室技术测定红细胞沉降率(erythrocyte sedimentation rate, ESR)、C反应蛋白(C-reactive protein, CRP)、免疫球蛋白A(Immunoglobulin A, IgA)、免疫球蛋白M(Immunoglobulin M, IgM)、免疫球蛋白G(Immunoglobulin G, IgG)、补体C3、补体C4、类风湿因子(rheumatoid factor, RF)、抗核抗体(antinuclear antibody, ANA)、抗着丝点抗体(anti-centromere antibodies, ACA)、抗心磷脂抗体(anticardiolipin, aCL)、抗双链DNA抗体(抗dsDNA抗体)、抗Sm抗体、抗RNP抗体、抗Scl-70抗体、抗Ro-52抗体、抗SSA抗体、抗SSB抗体、着丝点蛋白A(centromere protein A,CENP-A)、着丝点蛋白B(centromere protein B,CENP-B)等指标。记录雷诺现象和改良的Rodnan评分(modified Rodnan skin score, MRSS)等临床表现来评估SSc的疾病状态,分析抗tubulin-α-1C抗体与其他实验室指标和临床表现的关系。组间计量资料比较采用两独立样本t检验或Mann-Whitney U检验;组间计数资料比较采用卡方检验;采用绘制实验组工作曲线确定抗tubulin-α-1C抗体对诊断SSc最佳截断值并分析其诊断效能,相关性分析采用Spearman相关分析。结果:SSc患者、SLE患者、pSS患者和正常对照组血清中抗tubulin-α-1C抗体水平分别为81.24±34.38、87.84±38.52、59.79±25.24、39.37±18.7,SSc患者血清抗tubulin-α-1C抗体水平显著高于pSS患者和正常对照组,P均小于0.001。SSc患者血清抗tubulin-α-1C抗体水平与SLE组差异没有统计学意义。Spearman相关性分析示抗tubulin-α-1C抗体与SSc炎症和疾病活性标志物ESR正相关(r=0.313,P=0.019),与MRSS正相关(r=0.636, P<0.01)。根据正常对照组表达抗tubulin-α-1C抗体$\bar{x}+2s$定义阳性阈值为76.77,将SSc患者分为抗tubulin-α-1C抗体阳性组和阴性组,抗tubulin-α-1C抗体阳性组出现雷诺现象的比例明显高于阴性组(71.4% vs. 37.5%,P=0.039); 抗tubulin-α-1C抗体阳性组中抗Scl-70抗体、ACA抗体和aCL抗体的阳性率也显著高于阴性组(分别为37.9% vs. 15.2%,P=0.041; 34.5% vs. 12.1%, P=0.035; 13.8 vs. 0, P=0.027)。结论:抗tubulin-α-1C抗体在SSc患者血清中异常升高,可能成为一种新的生物标志物用于SSc的临床诊断和风险预测。

关键词: 系统性硬化症, 抗α-1C微管蛋白抗体, 自身抗体

Abstract:

Objective: To detect the serum level of a novel autoantibody, anti-tubulin-α-1C, in patients with systemic sclerosis (SSc) and to investigate its clinical significance. Methods: Anti-tubulin-α-1C antibody levels were determined by enzyme-linked immunosorbent assay (ELISA) in 62 patients with SSc, 38 systemic lupus erythematosus (SLE), 24 primary Sj?gren’s syndrome (pSS) patients, and 30 healthy controls (HCs). Erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), immunoglobulin A(IgA), immunoglobulin M (IgM), immunoglobulin G (IgG), C3, C4, rheumatoid factor (RF), antinuclear antibody(ANA), anti-centromere antibodies( ACA), anticardiolipin (aCL), anti-dsDNA antibody, anti-Sm antibody, anti-RNP antibody, anti-Scl-70 antibody, anti-Ro52 antibody, anti-SSA antibody, anti-SSB antibody, centromere protein A(CENP-A), centromere protein B (CENP-B) were measured by standard laboratory techniques. Raynaud’s phenomenon and modified Rodnan skin score(MRSS) were recorded to evaluate the disease status of SSc. Independent sample t test, Chi square test, Mann-Whitney U test, Spearman rank correlation were used for statistical analyses. Results: The serum anti-tubulin-α-1C antibody concentration in SSc group was 81.24±34.38, the serum anti-tubulin-α-1C antibody concentration in SLE group was 87.84±38.52, the serum anti-tubulin-α-1C antibody concentration in pSS group was 59.79±25.24, and the serum anti-tubulin-α-1C antibody concentration in healthy group was 39.37±18.7. Multivariate analysis revealed that anti-tubulin-α-1C antibody levels were significantly increased in the SSc and SLE patients. The expression level of anti-tubulin-α-1C antibody in SSc was higher compared with the pSS group and the health control group (P<0.01). Further analysis demonstrated that the elevated anti-tubulin-α-1C antibody were correlated with the SSc inflammation and disease activity markers ESR(r=0.313, P=0.019), The levels of anti-tubulin-α-1C antibody were also significantly correlated with MRSS(r=0.636, P<0.01). The best cut-off value for the diagnose of SSc was 76.77 as mean+2SD value. The proportion of Raynaud's phenomenon was higher in the group of anti-tubulin-α-1C autoantibody-postive SSc patients than that in anti-tubulin-α-1C autoantibody negative group(71.4% vs. 37.5%, P=0.039). The proportions of anti-Scl-70 antibody, anti-CENP antibody and anti-cardiolipin antibody were higher in the group of anti-tubulin-α-1C autoantibody-postive SSc patients than in the anti-tubulin-α-1C autoantibody negative group (37.9% vs. 15.2%, 34.5% vs. 12.1%, 13.8 vs. 0, respectively, all P<0.05). Conclusion: Based on this explorative stu-dy, the level of anti-tubulin-α-1C antibody increased in the serum of the patients with SSc. There were correlations between anti-tubulin-α-1C autoantibody and clinical and laboratory indicators of the SSc patients. It may become a novel biomarker indicative of active SSc and could be applied in future clinical practice.

Key words: Systemic sclerosis, Anti-tubulin-α-1C antibody, Autoantibodies

中图分类号:

R783.2

赵静,孙峰,李云,赵晓珍,徐丹,李英妮,李玉慧,孙晓麟. 抗α-1C微管蛋白抗体在系统性硬化症中的表达及临床意义[J]. 北京大学学报（医学版）, 2020, 52(6): 1009-1013.

Jing ZHAO,Feng SUN,Yun LI,Xiao-zhen ZHAO,Dan XU,Ying-ni LI,Yu-hui LI,Xiao-lin SUN. Significance of anti-tubulin-α-1C autoantibody in systemic sclerosis[J]. Journal of Peking University (Health Sciences), 2020, 52(6): 1009-1013.

图/表 2

表1

抗tubulin-α-1C抗体阳性组与阴性组间实验室指标的比较"

Items	Anti-tubulin-α-1C(+)(n=29)	Anti-tubulin-α-1C(-)(n=33)	P
Female, n(%)	26 (89.7)	31 (93.9)	0.537
Age/years, $x - ± s$	59.07±20.42	51.88±10.45	0.081
ESR /(mm/h), M(Min, Max)	21 (7-110)	12 (2-55)	0.033^*
CRP/(mg/L), M(Min, Max)	2.49 (0.54-73.90)	2.95 (0.38-44.21)	0.431
C3/(g/L), $x - ± s$	0.87±0.23	0.87±0.26	0.966
C4/(g/L), $x - ± s$	0.22±0.09	0.24±0.26	0.667
RF/(IU/mL), M(Min, Max)	20 (20-123)	20 (18-3 710)	0.781
Anti-Ro52 positive, n(%)	8 (27.6)	16 (48.5)	0.090
Anti-RNP positive, n(%)	7 (24.1)	9 (27.3)	0.780
Anti-Scl-70 positive, n(%)	11 (37.9)	5 (15.2)	0.041^*
ANA positive, n(%)	23 (79.3)	20 (60.6)	0.110
ACA positive, n(%)	10 (34.5)	4 (12.1)	0.035^*
CENP A or B positive, n(%)	6 (20.7)	6 (18.2)	0.803
aCL positive, n(%)	4 (13.8)	0	0.043^*
Anti-dsDNA positive, n(%)	4 (13.79)	1 (3.03)	0.120
IgA/(g/L), $x - ± s$	3.37±2.89	2.52±1.80	0.164
IgG/(g/L), $x - ± s$	15.30±6.61	14.60±6.82	0.681
IgM/(g/L), $x - ± s$	1.17±0.81	0.99±0.70	0.350
Raynaud’s phenomenon, n(%)	15 (71.4)	6 (37.5)	0.039^*

表1

表2

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Items	r	P
Age/years	0.023	0.860
Disease duration	0.118	0.455
ESR	0.313	0.019^*
CRP	-0.109	0.432
IgA	0.121	0.350
IgG	0.168	0.193
IgM	0.114	0.384
C3	-0.042	0.744
C4	0.086	0.513
RF	-0.079	0.541
aCL	0.330	0.106
Anti-dsDNA	0.146	0.477
MRSS	0.636	<0.001^*