Abstract
Expression of the “protective genes” A20, heme-oxygenase-1, and Bcl-xl in rodent allografts and xenografts correlates with long term survival of transplanted hearts. To date, there has been no published reports of expression of these molecules following human cardiac transplantation. Here we have investigated expression of A20, HO-1 and BCL-2 in sequential biopsies from 9 cardiac transplant recipients using quantitative real time RT-PCR. The aim of the study was to investigate the temporal expression after transplantation and investigate whether mRNA levels correlate with acute rejection (AR). 5-16 biopsies were analysed from each of 9 patients, from 7 to 600 days after transplantation ( 98 biopsies in total). Biopsies were classified as showing AR if they showed grade 1A or above. In order to compare mRNA values from one biopsy to another, all values were normalised against a single value (obtained at the earliest point, day 7 in one patient without rejection) . The results were calculated for each patient from 0-2, 3-9 and 10-24 months after transplantation. All three genes were expressed at every time point compared to the endogenous control gene. HO-1 was significantly higher in the first two months (mean +/-SEM:, 1.3+/-0.4 at 2 months vs 0.5+/-0.1 at 10 months +, p < 0.05). In contrast expression of A20 and BCL-2 is low at 2 months but significantly increases with time (p < 0.05 2 months vs 10 months +). There was significantly more expression of HO-1 in AR (1.0+/-0.2) versus non-rejection (0.5+/-0.1, p = 0.037); in contrast, there was no significant association of A20 or BCL-2 with AR. Studies are currently underway to investigate immunocytochemical localisation of these proteins in the biopsies. In conclusion, HO-1 is highly expressed in the first two months : this may reflect a response to trauma or AR. Further studies are under way to investigate whether early expression correlates with protection from development of chronic graft vasculopathy.
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