Preparation and purification of horse antihuman lymphocyte globulin (ALG)

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IN recent years great interest has been aroused in the immunosuppressive activity of antilymphocyte serum (ALS) or antilymphocyte globulin (ALG). The immunosuppressive activity of ALG has been demonstrated in various experimental models, and it has been claimed that ALG may be the most potent immunosuppressive agent yet known. ALG has also proved to be helpful as an adjunct in the immunosuppressive regimen for human renal transplantation. Starzl and associates1 reported that with use of ALG prepared by the ammonium sulfate fractionation method there was 95 percent one-year survival among patients who underwent renal transplantation. The major complications of ALG therapy noted by these authors included severe pain at the site of ALG injection, and a low but significant incidence of anaphylactic reactions.

Recently we investigated the problems of the preparation, purification, and clinical use of horse antihuman lymphocyte globulin as related to the Cleveland Clinic renal transplantation program. By using a DEAE-Sephadex batch fractionation method for the purification of ALG we have been able to obtain the IgG component of horse serum with a more than 95 percent purity. We have used this material in the treatment of 41 patients during the last year; this report summarizes our experience with respect to the methods of preparation and purification of ALG.

Materials and Preparation of ALG

Spleen cell antigen. The antigenic preparation used for the immunization of horses consisted of a spleen cell suspension prepared in the following manner under sterile conditions. Excised human spleens, of patients with terminal renal failure . . .



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