Abstract
BackgroundVariants of the interferon-lambda3 (IFNL3) gene have been associated with both spontaneous and treatment induced clearance of HCV infection. Attempts to link polymorphisms of the IFNL3 gene with variation in the level of IFNL3 expression have been inconclusive. This is partially due to the difficulty to design assays distinguishing IFNL3 from IFNL2.MethodsIn this study an allele specific real-time PCR (RT-PCR) assay was developed which allows the relative quantification of the two IFNL3 transcripts in cells heterozygous for SNP IFNL3.rs4803217 in the 3'UTR of the IFNL3 gene. This SNP is in strong linkage disequilibrium (LD) with the predictive marker rs12979860.ResultsRaji cells showed two-fold increased levels of IFNL3.rs4803217 C-allele expression. In peripheral blood mononuclear cells (PBMCs) of eight uninfected donors, two donors showed increased IFNL3.rs4803217 C-allele expression.ConclusionThis indicates that allele specific differences in IFNL3 expression vary between individuals and might contribute to the variety of outcomes in HCV infected patients.Electronic supplementary materialThe online version of this article (doi:10.1186/s12881-014-0104-7) contains supplementary material, which is available to authorized users.
Highlights
Variants of the interferon-lambda3 (IFNL3) gene have been associated with both spontaneous and treatment induced clearance of Hepatitis C Virus (HCV) infection
The relative amount of allele specific transcript was measured after interferon stimulation of Huh7, Raji and Jurkat cells, and in peripheral blood mononuclear cells (PBMCs) of eight uninfected healthy donors, which were heterozygous for IFNL3.rs4803217
We report that the presented IFNL3 specific assay is able to accurately measure variation of allele specific expression between individuals
Summary
Variants of the interferon-lambda (IFNL3) gene have been associated with both spontaneous and treatment induced clearance of HCV infection. Attempts to link polymorphisms of the IFNL3 gene with variation in the level of IFNL3 expression have been inconclusive. This is partially due to the difficulty to design assays distinguishing IFNL3 from IFNL2. Genetic polymorphisms in the interferon-lambda gene (IFNL3), known as interleukin 28B (IL-28B) have been shown to be highly associated with the outcome and treatment response of Hepatitis C infection in ethnically diverse cohorts [1,2,3,4,5,6]. We report that the presented IFNL3 specific assay is able to accurately measure variation of allele specific expression between individuals
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