Pretreatment of renal transplants with anti-CD45 antibodies: optimization of perfusion technique

Pig kidneys were perfused ex vivo with a monoclonal anti-pig CD45 antibody to determine optimum conditions for human kidney anti-CD45 perfusion, as such pretreatment may prolong renal allograft survival. Efficacy of perfused antibody uptake was assessed by immunohistochemical double-labelling of multiple renal biopsies. Antibody uptake by CD45+ cells was rapid, and maximal within one hour of perfusion. An antibody concentration of 40 micrograms/ml more effectively saturated available CD45 binding sites than lower concentrations. Whilst cortical perfusion was homogeneous (median uptake > 99%, minimum 90.0%) after 25, 50 or 100 ml perfusion with 40 micrograms/ml monoclonal antibody (mAb), the medulla was less evenly perfused with 25 ml (median uptake 82.6%) than with 50 or 100 ml (median uptake 98.5% and 98.7%, respectively). Prolongation of renal cold ischaemia of up to 46 hours prior to mAb perfusion did not adversely affect antibody binding. Clamping of the renal vein during and after mAb perfusion was associated with better cortical and medullary mAb uptake than if the vein was left unclamped. These data show that the vast majority of the kidney's CD45+ antigen-presenting cells can be bound by anti-CD45 mAb perfused ex vivo, if an adequate volume and concentration of mAb perfusate is employed and the renal vein is clamped during and after the process.