ABSTRACT
BACKGROUND: Accumulating evidence indicates that multiple genetic factors are involved in the pathogenesis of primary biliary cirrhosis (PBC). The aim of this study was to investigate whether polymorphisms of the integrin αV subunit gene (ITGAV), a component of integrin αVß6, which plays an important role in the process of fibrosis, are associated with susceptibility to the onset and/or progression of PBC. METHODS: In the primary study, eight tag single nucleotide polymorphisms (SNPs) in ITGAV were analyzed by polymerase chain reaction (PCR)-restriction fragment length polymorphism, direct DNA sequencing, or high-resolution melting curve analysis in 309 Japanese patients with PBC who were registered in the National Hospital Organization Study Group for Liver Disease in Japan (PBC cohort I) and 293 gender-matched healthy Japanese volunteers (control subjects). For the replication study, 35 PBC patients who progressed to end-stage hepatic failure and underwent liver transplantation (PBC cohort II) were also analyzed. RESULTS: Three tag SNPs (rs3911238, rs10174098, and rs1448427) in ITGAV were significantly associated with the severe progression of PBC, but not with susceptibility to the onset of PBC, in the primary study (PBC cohort I). Among these SNPs, rs1448427 was also significantly associated with the severe progression to end-stage hepatic failure in the replication study of PBC patients who underwent liver transplantation (PBC cohort II). CONCLUSIONS: ITGAV is a genetic determinant for the severe progression of PBC in Japanese patients. Genetic polymorphisms of ITGAV may be useful for identifying high-risk Japanese PBC patients, including those who will require liver transplantation, at the time of initial diagnosis.
Subject(s)
End Stage Liver Disease/etiology , Integrin alphaV/genetics , Liver Cirrhosis, Biliary/genetics , Adult , Aged , Asian People/genetics , Case-Control Studies , Disease Progression , End Stage Liver Disease/genetics , End Stage Liver Disease/therapy , Female , Genetic Predisposition to Disease , Humans , Japan , Liver Transplantation , Male , Middle Aged , Polymorphism, Restriction Fragment Length , Polymorphism, Single Nucleotide , Sequence Analysis, DNAABSTRACT
Green tea is an acknowledged cancer preventive in Japan, and the main constituent of green tea catechins is (-)-epigallocatechin gallate (EGCG). To investigate the bioavailability of EGCG in humans, we generated a monoclonal antibody against EGCG in BALB/c mice by immunizing thyroglobulin-conjugated EGCG. Out of 32 hybridoma cell lines, three hybridomas were selected by enzyme-linked immunosorbent assay (ELISA), and then determined by surface plasmon resonance assay: One hybridoma TG38 produced a specific monoclonal antibody against EGCG. The primary structure of TG38 light chain was then deduced from DNA sequence of the light chain gene. The NCBI-BLAST search showed the uniqueness of TG38 monoclonal antibody, and three amino acid residues specific for TG38 were aligned on two loops and one beta-sheet of the tertiary structure of the antibody. The TG38 antibody is the first monoclonal antibody against EGCG and catechins, since it bound to four green tea catechins with a galloyl group.
