In vitro reactivity of human lymphocytes in chronic uraemia: analysis and interpretation

Altered cellular immunity complicating chronic uraemia includes lymphocytopenia, thymic atrophy, impaired allograft rejection and delayed hypersensitivity in skin tests, diminished appearances of lymphocytes on skin windows, and shortened in vitro survival of uraemic lymphocytes. Studies were undertaken to further characterize these lymphocyte defects. Lymphocytes separated from peripheral blood of twenty-four uraemic patients were compared with those from fifty-one normal persons in their rates of RNA and DNA synthesis in both PHA-stimulated and unstimulated cultures, as determined by incorporation of radiolabelled precursors. Synthesis, expressed as absolute incorporation/10 6 viable lymphocytes, was accelerated in the majority of both stimulated and unstimulated uraemic cultures. Serial studies over several months of twenty-five uraemic patients (fifteen maintained by haemodialysis, ten by renal allotransplantation), compared with nineteen normal controls, showed these differences to be consistent and persistent. While normal lymphocytes exhibited stability, uraemic cells fluctuated widely in their serial synthesis rates. In 25% of such cultures, unstimulated synthesis exceeded that induced by PHA. Increased synthesis without PHA, reflecting enhanced spontaneous blastogenesis, is compatible with decreased survival, numbers, and functional capacity of uraemic lymphocytes. Reports by others of diminished PHA-responsiveness of uraemic lymphocytes are based upon whole-culture incorporation and/or expressed as ratios between stimulated and unstimulated cultures. Both shortened survival and accelerated spontaneous nucleic acid synthesis by uraemic lymphocytes cause such ratios to be misleadingly low. Such factors as cell numbers and viability at harvest, counting efficiency, and culture sterility are essential to avoid misinterpretations of such data based on incorporation of radiolabelled precursors to the nucleic acids.