Comparative activities of rabbit complements of different ages using an in-vitro marrow purging model

In order to determine the best time to obtain complement for use in marrow purging in vitro, we have used 51Cr-release and limiting dilution assays (LDA) to evaluate the ability of serum from rabbits 20-30 (25d), 30-40 (35d) and 40-50 (45d) days old to lyse a series of neoplastic target cells in the presence of complement activating antibodies. Utilizing a limiting dilution assay (LDA) to measure log depletion of tumor cells in a 20-fold excess of normal bone marrow, treatment with monoclonal antibody and 25d complement depleted at least 4 logs of leukemia or lymphoma cells. 35d and 45d complements were approximately one log less effective. When normal bone marrow alone was treated with either J5 (CD10) antibody alone, each complement alone or a combination of J5 antibody with each complement, there was no significant depletion of hematopoietic progenitors in any subgroup. These results suggest that complement from 20-30 day old rabbits should be used for the purpose of ex-vivo purging due to its highly sensitive and specific activity.