| Part 1 Polymorphism in chemokine CCL3L1,CCL4L1 gene and acute rejection and recurrence of hepatitis B in Chinese liver transplant recipientsAim:Copy number variation (CNV) in gene was a novel type of genetic diversity in human that affect the pathogenesis, progress and prognosis of disease. The aim of this study was to determine the associations between CNV in CCL3L1 gene and the rejection risk in liver transplant recipients.Methods:The 266 Han-Chinese patients and 185 recipients with HBV related end stage liver disease were enrolled in this study, Genomic DNA and mRNA samples were obtained from the whole peripheral blood, a quantitative real time PCR based assay was done to determine the copy number of CCL3L1, real time PCR was done to calculate the CCL3L1/CCL3 mRNA ratio. SNap-Shot technology was used to detect the rs1801628, rs17850251(CCL3L1), rs3744595, rs4796195(CCL4L1) single nucleotide polymorphism. Results:The CNV distributions of both CCL3L1 and CCL4L1 in patients and health controls had no significant differences. Copy number of CCL3L1 and CCL4L1 in acute rejection group (AR) shifted to higher number compared to non-acute rejection group (4.74±1.87 vs 3.88±2.13, p=0.017). Moreover, patients with higher CCL3L1 copies (?3 copies) had a significantly higher rejection risk than patients lower copies CCL3L1 (OR=3.85,95% CI,1.14-13.04; p=0.021). No association was found between CNV and rejection grades. No association was found between CNV and recurrence of hepatitis B, No associations was found between the SNP in CCL3L1 and CCL4L1 with acute rejection and recurrence of hepatitis BConclusion:liver transplant recipients with high copy number of CCL3L1 gene had a significant higher risk of acute rejection. CNV might be a novel genetic diversity that correlated with allograft rejection.Part 2 polymorphism in CCR5 gene and acute rejection and recurrence of hepatitis B in Chinese liver transplant recipientsAim:The aim of this study was to investigate the associations between polymorphisms in CCR5 gene and acute rejection in liver transplantation.Method:266 patients, who underwent liver transplantation from January 2006 to march 2009, were enrolled in this study. Genomic DNA was extracted from the whole blood, CCR5?32 was detected bay a sizing PCR method, eight nucleotide polymorphism (rs2734648, rs1799863, rs1799987, rsl799988, rs1800023, rs1800024, rs1800452, rs746492) loci in CCR5 gene were detected by Applied Biosystems SNaP-Shot technology.Results:CCR5A32 mutation was not detected in all the individuals from China. There was no significant association between the SNP in CCR5 gene and acute rejection nor recurrence of hepatitis B after liver transplantation.Conclusion:SNP in a single gene of CCR5 might not play a role in acute rejection and recurrence of hepatitis B after liver transplantation.Part 3 Combination of copy number variation in CCL3L1 gene with nucleotide polymorphism in CCR5-2459 and recurrence of hepatitis B in Chinese liver transplant recipientsaim:genetic diversity of chemokine and chemokine receptor had been reported to be associated with the outcome of hepatitis B virus infection. The aim of this study was to evaluate whether the copy number variation (CNV) in CCL3L1 gene and SNP in CCR5-2459A>G(rs1799987) were associated with recurrent hepatitis B in liver transplantation for hepatitis B virus infection related end stage liver disease.Methods:A total of 266 transplant recipients among which 185 diagnosed as end stage liver disease were enrolled in this study, the genomic DNA was extracted from the whole blood, the copy number of CCL3L1 gene was detected by a quantitative real time PCR based assay, and the single nucleotide polymorphism in CCR5 rs1799987 was detected by SNaP-Shot technology.Results:neither copy number variation nor polymorphism in rs1799987 was associated with post transplant re-infection of HBV. However, lower copies (<4) CCL3L1 gene compared to population median in combination with CCR5 G allele had a significant higher risk for recurrent hepatitis B (odd ratio=1.93,95% CI:1.00-3.69; P=0.047). SNP in rs1799987 were not associated with post-transplantation acute rejection. However the higher copy number in CCL3L1 was associated with rejection risk, patients with compound high CCL3L3L1 copy number and rsl799987 A allele were associated with higher rejection risk after liver transplantation.Conclusion:copy number variation was a novel genetic diversity of human, which correlated with biological effect.Conclusion:patients possessed the compound decreased functional genotype of both CCL3L1 and CCR5 gene might be favor of recurrence of hepatitis B after transplantation. |
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