Correlation between IL28B/TLR4 genetic variants and HCC development with/without DAAs treatment in chronic HCV patients

In Egypt,Sofosbuvir(SOF)in combination with Dataclasvir(DCV)is the broadly used DAAs with excellent therapeutic profile.This study is designed to explore the relation between IL28B/TLR4 genetic variants and each of the followings;HCC development post SOF/DCV treatment,progression to HCC in naı¨ve patients and SOF/DCV therapy outcome.A total of 493 blood samples were collected(controls(n = 70);HCV patients treated with SOF/DCV(n = 252)of whom 65 patients developed HCC,187 patients didn’t develop HCC(125 responders,62 relapsers);naı¨ve HCV patients(n = 171)had early(n = 48),late liver fibrosis(n = 21)and HCC(n = 102)).Both SNPs were genotyped using a TaqMan 50 allelic discrimination assay.At IL28B rs12979860 SNP,the C allele was significantly correlating with the response rate more than T allele(OR 1.9,95%CI 1.29e2.9,p=0.004),while at TLR4 rs4986791 SNP,no association was found(OR 6.5,95%0.57e75.28,p = 0.09).Both SNPs couldn’t detect the probability for HCC emergence after treatment.In naı¨ve patients,the protective alleles were detected in their lowest frequency in HCC patients(p = 0.1,for rs12979860 and,p = 0.001 for rs4986791).SOF/DCV combination improved SVR rates in HCV genotype 4a infected patients regardless of IL28B genotype,with the best rates in those lacking the T allele.

HCV Therapy Follow-up Fractionation (CTF2) by Intra-PBMC Nested RNA PCR Recognizes Early Virologic Response and Relapse

Background and Aims:Sustained virologic response is evaluated by single-step reverse transcription(SRT)PCR as-say,which assesses hepatitis C virus(HCV)clearance from plasma but not from tissues such as peripheral blood mono-nuclear cells(PBMCs).Persistence of HCV RNA in PBMCs beyond end of treatment(EOT)is associated with nonres-ponse.Our goal was to measure intra-PBMC HCV RNA levels during oral antiviral therapy according to the HCV therapy follow-up fractionation(CTF2)protocol.Methods:Compen-sated chronic HCV patients(n=278 SRT-PCR positive)were scheduled to receive oral antiviral therapy.Subjects were followed-up by SRT and intra-PBMCs HCV RNA PCR at the end of the 2nd,6th,10th,14th,18th and 24th weeks to evaluate virus clearance from plasma and PBMCs,respectively.The CTF2 protocol evaluated SRT and PBMC PCR status at each follow-up point for determining therapy continuation or inter-ruption to address cost effectiveness.Results:All patients tested negative by SRT PCR after therapy for 2 weeks.Appli-cation of the CTF2 protocol revealed:a)increasing HCV clear-ance rate from 75.9%at the end of 10th week to 90.3%at the end of 24th week(p<0.00001);b)faster clearance of HCV from plasma compared to PBMCs at each point of follow-up until the 18th week(p<0.05);c)higher viral elimination rates diagnosed by PBMC HCV RNA PCR(?)compared to PBMC HCV RNA PCR(+)from the 6th to 24th week of treatment(p<0.0001);d)higher over-time increase curve of combined plasma and PBMC HCV RNA determined negativity compared to the decline in positivity curves by PBMC PCR at the 6th-18th week compared to the 24th week(p<0.01)—these results validated treatment continuation;and e)solitary evaluation of EOT sustained HCV infection and relapses by PBMC HCV RNA(p<0.001).Conclusions:Early elimination of serum and tissue(PBMC)HCV infection by oral antiviral therapy can be achieved and evaluated during a cost-effective CTF2 pro-tocol application.