Division of Hepato-Gastroenterology, Department of Internal Medicine, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, Taiwan.
College of Medicine, Chang Gung University, Taoyuan, Taiwan.
Biomed Res Int. 2018 Mar 4;2018:1826140. doi: 10.1155/2018/1826140. eCollection 2018.
The aim of this study is to elucidate the biogenetic modification of donor and recipient interleukin-28B (IL-28B) genotypes in liver graft biopsies after living donor liver transplantation (LDLT) for chronic hepatitis C virus- (HCV-) related, end-stage liver disease. Fifty liver graft biopsies were collected from recipients during LDLT treatment for HCV-related, end-stage liver disease. DNA was extracted from all 50 liver tissues, and the IL-28B single-nucleotide polymorphisms (SNPs) rs8099917 and rs12979860 were studied for allelic discrimination by real-time PCR analysis. Blood samples were obtained from donors and recipients on postoperative day 0 (POD0), POD7, and POD30. We randomly selected five liver biopsies and isolated the hepatocytes by laser capture microdissection (LCM) to evaluate genotype modifications resulting from LDLT. After LDLT, the IL-28B SNP rs8099917 was identified not only in the liver graft biopsies and donors' sera (TT = 41 : 43; GT = 9 : 5; GG = 0 : 2), but also in liver graft biopsies and recipients' sera on POD0 (TT = 41 : 44; GT = 9 : 4; GG = 0 : 2), POD7 (TT = 41 : 30; GT = 9 : 18; GG = 0 : 2), and POD30 (TT = 41 : 29; GT = 9 : 19; GG = 0 : 2). A significant difference was observed between the rs8099917 allele frequencies of liver graft biopsies and recipients' sera on POD30 ( = 0.039). In addition, a significant difference was also noted between the rs12979860 allele frequencies of liver graft biopsies and donors' sera (CT = 49 : 39; TT = 1 : 10) ( = 0.012) and of liver graft biopsies and recipients' sera on POD0 (CT = 49 : 39; TT = 1 : 11) ( = 0.002), POD7 (CT = 49 : 42; TT = 1 : 8) ( = 0.016), and POD30 (CT = 49 : 41; TT = 1 : 9) ( = 0.008). This phenomenon was confirmed by pyrosequencing of hepatocytes isolated by LCM. Following LDLT, the TT-to-GT IL-28B genotype modification predominated in rs8099917, and the CC-to-CT modification predominated in rs12979860. In conclusion, these modified phenomena suggested that the selected donor with a predictable and favourable IL-28B genotype will not confer a benefit on the recipient in the living donor liver transplantation setting.
本研究旨在阐明在活体肝移植（LDLT）治疗丙型肝炎病毒（HCV）相关终末期肝病后，供体和受者白细胞介素 28B（IL-28B）基因型在肝移植物活检中的生物遗传修饰。在 LDLT 治疗 HCV 相关终末期肝病期间，从 50 名受者的肝组织中收集了 50 份肝移植物活检样本。从所有 50 个肝组织中提取 DNA，并通过实时 PCR 分析研究 IL-28B 单核苷酸多态性（SNP）rs8099917 和 rs12979860 的等位基因鉴别。在术后第 0 天（POD0）、第 7 天（POD7）和第 30 天（POD30）时从供者和受者获得血样。我们随机选择五个肝活检样本，并用激光捕获微切割（LCM）分离肝细胞，以评估 LDLT 引起的基因型修饰。在 LDLT 后，在肝移植物活检样本和供者的血清中（TT = 41:43；GT = 9:5；GG = 0:2）不仅发现了 IL-28B SNP rs8099917，而且在肝移植物活检样本和受者的血清中（TT = 41:44；GT = 9:4；GG = 0:2）在 POD0、POD7 和 POD30 时（TT = 41:30；GT = 9:18；GG = 0:2）也发现了这种情况。在 POD30 时，肝移植物活检样本和受者血清中的 rs8099917 等位基因频率存在显著差异（ = 0.039）。此外，在肝移植物活检样本和供者血清中的 rs12979860 等位基因频率（CT = 49:39；TT = 1:10）（ = 0.012）和肝移植物活检样本和受者血清中的 rs12979860 等位基因频率（CT = 49:39；TT = 1:11）（ = 0.002）、POD7（CT = 49:42；TT = 1:8）（ = 0.016）和 POD30（CT = 49:41；TT = 1:9）（ = 0.008）也存在显著差异。这一现象通过 LCM 分离的肝细胞的焦磷酸测序得到了证实。在 LDLT 后，rs8099917 中的 TT 到 GT 基因型修饰占主导地位，而 rs12979860 中的 CC 到 CT 修饰占主导地位。总之，这些修饰现象表明，选择具有可预测和有利的 IL-28B 基因型的供体并不能使受者在活体供肝移植环境中受益。
