Liver Unit, Hospital Clínic, CIBERehd, IDIBAPS, Barcelona, Spain.
J Hepatol. 2012 Feb;56(2):355-8. doi: 10.1016/j.jhep.2011.07.027. Epub 2011 Sep 1.
BACKGROUND & AIMS: Recipient and donor IL28B polymorphisms seem to play an important role in the response to hepatitis C treatment after liver transplantation (LT). Since donor peripheral blood mononuclear cells (PBMC) are not always available, the aim of our study was to assess whether follow-up biopsies obtained after LT could be used to determine donor IL28B genotype.
Genotyping of IL28B rs12979860 was performed by TaqMan real-time PCR and direct sequencing in 56 HCV-infected LT recipients and their donors. Liver biopsies were obtained at the moment of LT (reperfusion) and at any time when clinically indicated (follow-up). Direct sequencing always confirmed the real-time PCR results.
Genotyping of donor IL28B rs12979860 polymorphisms showed a 100% match both in PBMC and reperfusion biopsies. The frequency of IL28B rs12979860 polymorphisms differed significantly between donors and follow-up biopsies (p=0.024). We found an enrichment of the IL28B rs12979860 CT genotype (72%) in follow-up biopsies compared to donor samples (46%). Recipient alleles were clearly detected in 14 heterozygous follow-up samples: 10 CT/CC, 1 CT/TT, and 3 TT/CC (recipient/donor), thus reflecting a mixture of both donor and recipient genotypes.
Our results support that follow-up liver biopsies from LT recipients are not suitable for determining donor IL28B rs12979860 genotype by TaqMan real-time PCR or direct sequencing and that PBMC or reperfusion biopsies should be used instead. Thus, it is very important to obtain adequate samples in order to accurately determine the relative contributions of both donor and recipient.
受体和供体 IL28B 多态性似乎在肝移植（LT）后治疗丙型肝炎的反应中起着重要作用。由于供体外周血单核细胞（PBMC）并不总是可用，我们的研究目的是评估 LT 后获得的随访活检是否可用于确定供体 IL28B 基因型。
采用 TaqMan 实时 PCR 和直接测序法对 56 例 HCV 感染的 LT 受者及其供者的 IL28B rs12979860 进行基因分型。在 LT（再灌注）时以及临床需要时（随访）获得肝活检。直接测序总是确认实时 PCR 结果。
供体 IL28B rs12979860 多态性的基因分型在 PBMC 和再灌注活检中均显示出 100%的匹配。供体 IL28B rs12979860 多态性的频率在供体和随访活检之间存在显著差异（p=0.024）。与供体样本（46%）相比，我们发现随访活检中 IL28B rs12979860 CT 基因型（72%）明显富集。在 14 例杂合的随访样本中清楚地检测到受体等位基因：10 例 CT/CC、1 例 CT/TT 和 3 例 TT/CC（受体/供体），因此反映了供体和受体基因型的混合。
我们的结果支持 LT 受者的随访肝活检不适合通过 TaqMan 实时 PCR 或直接测序来确定供体 IL28B rs12979860 基因型，而应使用 PBMC 或再灌注活检。因此，获得足够的样本以准确确定供体和受体的相对贡献非常重要。
