Hugh O. O'Kane

The Distribution of Rabbit Anti-Mouse Thymus Globulin After Injection into Mice∗

The immunosuppressive action of heterologous antilymphoid sera (ALS) has been well established in the laboratory in various species of animal, and prolongation of survival of allografts of skin (1), kidney (2), and liver (3) and xenografts of tumor (4) has been achieved with their use. Such observations have led to the clinical trial of ALS as an adjunct to more conventional immunosuppressive therapy in the grafting of organs in man (5, 6). Although theories have been formulated as to the mechanism of the immunosuppressive action (7), no definitive explanation has been found. We considered that a study of the distribution of heterologous antilymphoid globulin in vivo after intravenous injection might be contributory in this regard and the distributions of radio labeled gamma globulin and IgG prepared from heterologous antithymus serum were investigated after their parenteral injection. Presented here is a report of the results of these experiments. Materials and Methods. Rabbit anti-mouse thymus sera (RAMTS) were prepared by injecting intravenously each of three New Zealand rabbits (average weight 2 kg) with suspensions of 1 × 109 viable thymus cells obtained from neonatal mice. Each rabbit was injected twice at a 2-weekly interval and bled 1 week after the second injection. The sera were pooled and heated at 56° for 30 min. The initial titer of lymphocytotoxicity in vitro for murine lymphocytes was 1:4096 when tested by the method described by Terasaki and associates (8) and titrated to less than 10% kill. After repeated absorptions with murine red cells (1 vol erythrocytes: 4 vol serum) to remove in vitro he-magglutinating and hemolyzing activity, and a single absorption with a homogenate of murine kidney (1 vol kidney homogenate: 4 vol serum), the gamma globulin (ATG) was precipitated with 40% ammonium sulfate. In preparation for labeling with radioactive iodine, one portion of the precipitate was reconstituted in sodium phosphate buffer, 0.05 M, pH 7.0, to the original volume of the serum and then dialyzed against the same buffer. At this stage the lymphocytotoxic titer of the globulin (ATG) was 1:4096 when tested in vitro against murine lymphocytes from the peripheral blood and titrated to less than 10% kill. After reconstitution in 0.1 M potassium phosphate buffer (pH 8.0) another portion of the precipitate was dialyzed against this same buffer prior to the separation of the IgG (ATIgG) by column chromatography using diethylaminoethanol (DEAE) cellulose (9). The immunoelectrophoretic pattern of this fraction showed the presence of a single band of precipitation located in the position normally ascribed to IgG.

Surgical Treatment of Lesions of the Outflow Tract of the Left Ventricle

Presented at the Nineteenth Annual Meeting of the American College of Angiology, San Juan, Puerto Rico, January 21-25, 1973. From the Department of Surgery, Washington University School of Medicine, and the Division of Thoracic and Cardiovascular Surgery, The Jewish Hospital of St. Louis, Missouri. Supported in part by Research Grant No. HL-13088 from the National Heart and Lung Institute, United States Public Health Service, Department of Health, Education and Welfare. Left ventricular outflow lesions have become amenable to accurate corrective surgery with the advent of diagnostic methods which permit differentiation of the various pathologic states that may be present, and with open-heart surgery which permits appraisal of the pathologic anatomy and precise correction under direct vision. Until recently, many of these conditions had been of academic interest only. Lesions of the outflow tract of the left ventricle may be congenital or acquired, and are more completely defined in connection with the following classification: 1. Obstructive lesions A. Valvular stenosis 1. Congenital valvular stenosis

Organization of a cardiac surgical unit.

From the Departments of Surgery, Washington University School of Medicine, and The Jewish Hospital of St. Louis, St. Louis, Missouri 63110. Supported by Army contract DADA-17-69-9165 and U.S. Public Health Service Grant 2 RO 1 HL 12278-04. Presented at the nineteenth annual meeting of the American College of Angiology, San Juan, Puerto Rico, January 21-25, 1973. * Edison Professor of Surgery, Washington University School of Medicine and Surgeon-inChief, Jewish Hospital of St. Louis, St. Louis, Missouri. t Assistant Professor of Surgery, Washington University School of Medicine and Director, Division of Thoracic & Cardiovascular Surgery, Jewish Hospital of St. Louis. ‡ Instructor in Surgery, Washington University School of Medicine and Assistant in Thoracic & Cardiovascular Surgery, Jewish Hospital of St. Louis, St. Louis, Missouri. The title of this presentation suggests a technical treatise on how to do it; how