川崎病的免疫遗传学初步研究

【作者】 杨军；

【导师】 李成荣；

【作者基本信息】 重庆医科大学 ， 儿科学， 2003， 硕士

【摘要】 目的： 川崎病（Kawasaki Disease, KD）主要病理改变为全身非特异性血管炎，其病因未明，免疫调节异常，免疫活性细胞过度激活可能是KD的免疫发病机制。免疫细胞过度激活产生的多种细胞因子介导的促炎与抗炎症反应失衡对川崎病的发生、发展和冠状动脉损伤起着重要作用。近年研究发现：细胞因子基因多态性可影响个体细胞因子产生水平和反应强度，系统性红斑狼疮、类风湿性关节炎等多种自身免疫性疾病与肿瘤坏死因子α（Tumor Necrosis Factor alpha，TNF-α）和白细胞介素10（Interleukin-10，IL-10）基因多态性密切相关，而二者与川崎病发病的关联，国内外尚无系统研究。本研究对TNF-α和IL-10基因启动子区域4个多态性位点进行分析，旨在探讨TNF-α和IL-10基因多态性与川崎病发病及临床表型的相关性。方法： 应用聚合酶链反应-限制性内切酶片段长度多态性分析（PCR-RFLP）方法检测TNF-α和IL-10基因启动子区4个多态性位点，对96例川崎病患儿及160例正常对照儿童进行相关分析。结果:（1）中国汉族儿童TNF-α基因-308（A/G）位点等位基因频率与日本人和美国白人相近；而IL-10基因-1082（G/A）、-819（C/T）、-592（A/C）3个位点等位基因频率与英国人有显著性差异（P均<0.01）；（2）TNF-α基因-308位点等位基因频率在川崎病组和健康对照组间无<WP=8>显著性差异；IL-10 -1082、-819、-592 3个位点等位基因频率和单体型在两组之间比较亦无显著性差异。（3）临床表型分析发现：TNF-α-308A基因型在静脉注射免疫球蛋白(IVIG)非敏感型川崎病患者中的发生频率高于TNF-α-308G基因型，其差异有显著性意义（67%vs5%，X=90.48，P<0.01），前者发生IVIG非敏感型川崎病的相对危险度(RR)是后者的42.25倍(RR 95%CI=15.81～112.88,P<0.01)。IL-10-1082A/-819T/-592A单体型在川崎病合并冠状动脉损伤患者中的发生频率高于明显高于Non-ATA单体型, 两组间有显著性差异(52%vs20%，X2=18.36, P<0.01), IL-10基因启动子区ATA单体型合并冠状动脉损伤的相对危险度(RR)是Non-ATA单体型的4.26倍(RR 95%CI=2.20～8.25,P<0.01)。结论：TNF-α基因启动子区-308位点多态性与川崎病IVIG疗效有关联，IL-10基因启动子区-1082/-819/-592单体型与川崎病冠状动脉损伤有关，IL-10基因启动子区-1082A/-819T/-592A单体型可作为川崎病冠状动脉损伤高危人群的基因标志。更多还原

【Abstract】 Objective：Kawasaki disease (KD) is an acute febrile vasculitic sydrome of unkown etiology. It has been suggested that cytokines like tumor necrosis factor alpha (TNF-α) and interleukin-10(IL-10) are key player during acute KD. Recently, the polymorphisms relative to major transcriptional start site of TNF-α and IL-10 gene were shown to influence the level of TNF-α and IL-10 production in vitro. This study was aimed to investigated the genetic association of TNF-α and IL-10 promoter polymorphisms in juvenile Hans nationality patients with KD, and to explore possible associations with clinical manifestations of the disease. Methods：Four polymorphisms sites of TNF-α and IL-10 gene promoter regions from 96 children with KD were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RELP). 160 age-matched normal children from the Han nationality were used as controls. All patients were performed Doppler echocardiogram -phy examination in order to differentiate coronary artery lesion. Results： (1)The allele frequencies of -308(A/G) site of the TNF-α gene have no significant difference among children from the Han nationality ,the Japanese and Caucasian in America(P>0.05); The allele frequencies of the sites -1082(G/A), -819(C/T) and -592(A/C) of IL-10 gene<WP=10>were significantly different between children from the Han nationality and their British Counterparts(P<0.01). (2) The allele frequencies of -308(A/G) site of TNF-α gene have no significant difference between children with KD and normal controls; The haplotypes and the allele frequencies of the three sites of IL-10 have no significant difference between the two groups. (3) When clinical features were examined, the genotype frequency of TNF-α-308A was significantly higher in IVIG-resistant KD patients than those IVIG-sensitive patients (56% vs 3%), the genotype of TNF-α-308A was closely associated with IVIG-resistant KD (P<0.01, odds ratio 42.25, 95% confidence interval 15.81～112.88). The haplotype frequency of IL-10 -1082A/-819T/-592A was also higher in patients with coronary artery lesion(CAL) caused by KD than those patients without CAL(82% vs 52%), the haplotypes of IL-10 -1082A/-819T/-592A was significantly associated with CAL caused by KD (P<0.01, odds ratio 4.26, 95%confidence interval 2.20～8.25) . Conclusion：The polymorphism at position -308A of TNF-α is the important factor that probably influence the therapy efficiency of KD. The haplotypes (-1082/-819/-592) of IL-10 gene promoter might be related to the coronary artery complication of KD and -1082A/-819T/-592A haplotypes might be regarded as a genetic marker of risk factor for coronary artery lesion in KD.更多还原