Allogeneic bone marrow transplantation: the monitoring of granulocyte macrophage colonies following the collection of bone marrow mononuclear cells and after the subsequent in-vitro cytolysis of OKT3 positive lymphocytes

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Abstract
Marrow nucleated cells from 8 normal allogeneic donors was layered on Ficoll-Metrizoate to isolate the mononuclear cell fraction. The cells were then washed to remove Ficoll-Metrizoate and coagulation factors prior to resuspension in a balanced salt solution and the addition of the murine anti-human T-lympocyte monoclonal antibody OKT3 and rabbit complement. The procedures were assessed for their effect on mononuclear cell viability (mean recovery 84.4%); the ability of the cells to proliferate granulocyte-macrophage colonies (mean recovery 57.4%); the in-vitro T-lymphocytolysis (mean 75.7%) and the removal of rabbit complement (> 99%). Following marrow transplantation with this treated mononuclear fraction the mean day to recovery of .gtoreq. 1.0 .times. 109/I leukocytes was 20 days, with 3 patients developing .gtoreq. grade II acute graft vs. host disease (GvHD). Thus, treatment of donor marrow with OKT3 and complement in a large volume system was not detrimental to subsequent engraftment, nor effective in complete T-lymphocytolysis, nor in prevention of severe GvHD.