Toll样受体4和CD_(14)基因多态性与川崎病易感性的相关研究

Association of Toll-Like Receptor 4 and CD_(14) Gene Polymorphisms with Kawasaki Disease Susceptibility

目的探讨Toll样受体4(TLR4)及脂多糖受体CD14基因多态性与川崎病(KD)易感性的关系。方法应用三色荧光标记流式细胞术检测76例KD患儿和118例健康儿童外周血白细胞TLR4表达水平,采用聚合酶链反应限制性片断长度多态性(PCR-RFLP)法检测2组儿童TLR4基因(-896A/G),(-1196C/T)位点和CD14基因(-260C/T)位点的基因型频率和等位基因频率及其与KD的关系。结果1.KD组和健康对照组外周血淋巴细胞、中性粒细胞、单核细胞TLR4平均免疫荧光强度(MFI)分别为2.87±0.96,10.55±4.87,23.36±8.28和3.26±0.65,7.55±1.21,25.41±6.97;2.在KD组和健康对照组均未发现TLR4基因(-896A/G)和(-1196C/T)的多态位点;3.KD组和健康对照组CD14基因(-260C/T)位点均有突变,其CC,CT,TT基因型分布分别为35.5%、30.3%、34.2%和38.1%、47.5%、14.4%,差异有统计学意义(χ2=11.62P<0.05);其C,T等位基因频率则分别为50.7%、49.3%和61.9%、38.1%,差异有统计学意义(χ2=4.76P<0.05);其等位基因频率的相对风险分析发现T等位基因携带者发生KD的风险是C等位基因的1.58倍。结论TLR4基因(-896A/G)和(-1196C/T)位点多态性与KD发病无关,CD14基因(-260C/T)多态性T等位基因与KD发病密切相关,其可能是KD发病的遗传学易感因素。

Objective To explore the association of Toll - like receptor 4 TLR4 and lipopolysaecharide receptor CD14 gene polymorphisms with Kawasaki disease （KD） susceptibility. Methods Three - color fluorescent staining flow - eytometry was used to detect the expression of TLR4 in peripheral blood white blood cell of 76 KD children and 118 healthy control group. The gene of TLR4 （-896A/G） , （-1196C/T） and CD14 （-260C/T） polymorphisms was identified by polymerase chain reaction- restriction fragment length polymorphisms ; and the relationship between genotype and KD was analyzed. Results 1. The values of mean fluorescence intensity （MFI） of TLR4 in peripheral blood white blood cell of the KD groups and the healthy control groups were 2.87 ± 0.96, 10.55 ± 4.87, 23.36 ± 8.28 and 3.26 ± 0.65, 7.55 ±1.21, 25.41 ± 6.97, respectively ; There was a gradual increase of these values on lymphocyte, neutrophilic leukocyte and mononuclear cell in both groups. 2. （ - 896A/G） , （ - 1196C/T） polymorphisms of TLR4 gene were not found in both groups. 3. The frequency of each genotype of CD14 gene （-260C/T） was 35.5% CC, 30.3% CT, 34.2%TT in KD group and 38.1% CC, 47.5% CT, 14.4% TT in healthy control group. The frequency of each genotype was significantly different in 2 groups （Х^2 = 11.62 P 〈 0.05 ）. The genotype frequency of C and T alleles were 50.7% , 49.3% and 61.9% , 38.1% in KD group and healthy control group. The difference between the 2 groups had a statistical significance （Х^2 = 4.76 P 〈 0.05 ）. The relative risk for KD in T allele carriers was 1.58 times as high as that in C allele carriers. Conclusions The gene of TLR4 （ - 896A/G） , （ - 1196C/T） polymorphisms are not associated with KD. CD14 gene （-260C/T） T allele is associated with KD, and it might be a genetie predisposing predictor for KD.