高通量测序分析肝移植急性排斥反应相关基因单核苷酸多态性位点的突变

摘要

目的 利用高通量测序分析肝移植术后急性排斥反应相关度最高的基因单核苷酸多态性(SNP)位点的突变情况.方法 收集同种异体原位肝移植术的68例受体外周血样本,根据有否发生急性排斥反应分为急性排斥组(13例)和非排斥组(55例).通过查阅文献,最终确定与排斥反应发生相关的44个SNP突变位点.以44个SNP位点作为检测靶点,用高通量测序分析对两组受体外周血样本进行检测,经生物信息学分析出与急性排斥反应发生相关基因SNP位点的突变率.结果 急性排斥组中的白细胞介素(IL)-10 TT基因型、T等位基因、AA基因型、A等位基因的SNP位点突变率明显高于非排斥组;急性排斥组中的细胞趋化因子受体(CCR)5 AG基因型的SNP位点突变率明显低于非排斥组,CCR5 GG基因型的SNP位点突变率明显高于非排斥组;急性排斥组中的IL-4 CT 基因型的SNP 位点突变率明显高于非排斥组,IL-4 TT 基因型的SNP 位点突变率明显低于非排斥组;急性排斥组中核因子-κB 抑制因子α(NF-κBIA)C 等位基因的SNP 位点突变率明显高于非排斥组;急性排斥组中维生素D 受体(VDR)CC 基因型和C 等位基因的SNP 位点突变率明显低于非排斥组,差异均有统计学意义(均为P<0.05).结论 高通量测序分析发现肝移植术后急性排斥反应相关基因中,其SNP 位点突变率较高的包括IL-10 TT 基因型、T 等位基因、AA 基因型、A 等位基因,CCR5 GG 基因型、AG 基因型,IL-4 CT 基因型、TT 基因型,NF-κBIA C 等位基因,VDR CC 基因型和C 等位基因.%Objective To analyze the mutation of single nucleotide polymorphism (SNP) loci in genes most significantly associated with acute rejection after liver transplantation by high-throughput sequencing. Methods Peripheral blood samples were collected from 68 recipients undergoing allogeneic orthotopic liver transplantation. According to the incidence of acute rejection, all patients were divided into the acute rejection group (n=13) and non-rejection group (n=55). Through the literature review, 44 mutant SNP loci associated with acute rejection were finally identified. Using 44 SNP loci as the detection targets, high-throughput sequencing analysis was performed to detect peripheral blood samples in two groups of recipients. Bioinformatics analysis revealed the mutation rate of the SNP loci of genes related to acute rejection. Results The mutation rate of SNP loci of the interleukin(IL)-10 TT genotype,T allele,AA genotype and A allele in the acute rejection group was significantly higher than that in the non-rejection group. The mutation rate of the SNP loci of the cell chemokine receptor(CCR)5 AG genotype in the acute rejection group was significantly lower than that in the non-rejection group,the mutation rate of the SNP loci of the CCR5 GG genotype in the acute rejection group was significantly higher than that in the non-rejection group.The mutation rate of the SNP loci of IL-4 CT genotype in the acute rejection group was significantly higher than that in the non-rejection group,the mutation rate of the SNP loci of IL-4 TT genotype in the acute rejection group was significantly lower than that in the non-rejection group. The mutation rate of the SNP loci of nuclear factor-kappa B inhibitor alpha(NF-κBIA)C allele in the acute rejection group was significantly higher than that in the non-rejection group.The mutation rate of the SNP loci of vitamin D receptor(VDR)CC genotype and C allele in the acute rejection group was significantly lower than that in the non-rejection group. Differences were statistically significant (all P<0.05). Conclusions High-throughput sequencing analysis shows the genes associated with acute rejection after liver transplantation.Among them,the mutation rate of the SNP loci is relatively high in IL-10 TT genotype,T allele, AA genotype,A allele,CCR5 GG genotype,AG genotype,IL-4 CT genotype,TT genotype,NF-κBIA C allele,VDR CC genotype and C allele.