Differential HFE allele expression in hemochromatosis heterozygotes

Background & Aims: Hereditary hemochromatosis is associated with C282Y homo zygosity. Some heterozygotes may also present with abnormal iron parameters . However, the precise role of H63D and C282Y mutations in iron overload is poorly understood, We investigated the level of expression of the mutated and unmutated HFE alleles in these heterozygous patients. Methods: We studi ed the expression of HFE messenger RNAs in peripheral blood mononuclear cel ls from 34 heterozygotes using reverse-transcription polymerase chain react ion (PCR) followed by enzymatic digestion or sequence analysis of the PCR p roducts, which allows relative quantification of mutated and unmutated tran scripts, HFE proteins were quantified by Western blotting in Epstein-Barr v irus-immortalized lymphocyte extracts from 2 C282Y and H63D homozygotes and a compound heterozygote, Results: (187C > G; H63D) mutated transcripts pre dominated in H63D and compound heterozygotes and the normal transcripts in C282Y heterozygotes. The amount of HFE protein was increased in the H63D ho mozygotes and the compound heterozygote compared with the C282Y homozygotes . In addition, we found a new mutation at codon 282 (C282S) associated with severe iron overload, Conclusions: We demonstrate the existence of differe ntial allelic expression of the HFE alleles, suggesting that the (187C > G; H63D) mutation plays a role in the disease expression in H63D heterozygote s, in particular when associated with environmental or host factors.