At week 2 and month 6, surveillance biopsies were performed, and 4-μm formalin-fixed, paraffin-embedded sections were prepared. First, hematoxylin and eosin (H&E) staining was performed to characterize graft rejection according to the Banff criteria by double-blind scoring [16]. In addition, the sections were stained with cytokeratin 19 (CK19) to observe biliary formation, stained with C4d to indirectly assess the severity of AMR, and immunostained with antibodies against human CD4, CD8, and CD20 to assess immune cell infiltration. At least three fields (×200 magnification) from each patient were randomly selected to calculate the mean number of positive cells. Archival tonsil sections were used as positive controls. Sections treated with only the secondary antibody and diaminobenzidine (no primary antibody) served as the negative controls. All of the primary antibodies were purchased from Abcam (USA), and the secondary antibody and diaminobenzidine were purchased from DAKO (USA).

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