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The study of pro-inflammatory cytokines produced in situ in heart allografts may help to understand the mechanisms of rejection and open new possibilities to control graft rejection.A total of 23 endomyocardial biopsies obtained from 16 transplanted patients treated with triple-drug therapy (azathioprine, prednisone, and cyclosporine) were studied. mRNA expression for tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, IL-10, IL-12, IL-15, transforming growth factor (TGF)-β, and β-actin was determined by reverse transcription polymerase chain reaction (RT-PCR) and Southern blotting. Semiquantitative analysis was done by establishing the ratio between densitometric integrated value of each cytokine with the β-actin and correlated with the histopathologic findings.Three groups of biopsies were determined according to the International Society for Heart and Lung Transplantation criteria: grade 0 (control group, n = 12), grade 1A (sub-clinical rejection, n = 6) and 'quilty effect' (n = 5). An increased expression of mRNA for TNF-α and IL-6 (p = 0.0091 and 0.0075, respectively) was found associated with rejection grade 1A episodes, mRNA for IL-1β was nonspecifically expressed in all the study groups, while IL-10 mRNA was not detected in any of the biopsies studied. mRNA for IL-12 and IL-15 was not associated with rejection. Interestingly, TGF-β was not detected in any of the biopsies with the 'quilty pattern'.The association of TNF-α and IL-6 mRNA in situ expression with mild histologically probed rejection episodes may be used in the monitoring of heart transplants.