of a given patient are composed exclusively of molecules having either type I or type II L chains (7,8).This report will present evidence that populations of isolated erythrocyte autoanfibodies exhibit a strikingly high frequency of L chain homogeneity. Some of these autoantibody populations bearing only one detectable L chain type may also display relative homogeneity in electrophoretic mobility.
Materials and MethodsHuman Antibodies.--For isolation of autoantibodies, bleedings from patients 2 were taken into 0.01 M Na2EDTA or ACD solution, a Eluates from 6 times washed RBC stromata were made by acidification (14) or by heating at 60°C for 30 minutes. 8 Anfi-Rho antibodies (anti-D) (anti-Rhl) (15) from individual donor sera 4 were reacted at 37°C with the 6 times washed RBC of a healthy donor whose phenotype was O R_ho (ccDee) (Rh: 1, --2, --3, 4, 5) (15). Eluates from the washed, sensitized RBC were made by acidification. These eluates displayed no reactivity with Orh (ccddee) (Rh: --1, --2, --3, 4, 5) RBC. None of the Rh antisera or elnates contained demonstrable saline agglutinating activity for O Rho RBC. Autoantibody and Rh antibody eluates were concentrated by negative pressure ultrafiltration through collodion membranes. The only immunoglobulin detectable by hemagglutination and immunodiffusion in the concentrated eluates was 7S 5'-globulin. 5Antigens.--Bence Jones (B J) proteins, the pathologic equivalent of normal L chains (16), were isolated from urines of myeloma patients, Sm. (type I) and S1. (type II), by precipitation in 55 per cent saturated ammonium sulfate followed by chromatography on DEAE cellulose (17). Highly purified 7S myeloma globulins were obtained by utilizing their properties as euglobulins or cryoglobulius. The original typing of BJ and myeloma proteins was established by Ouchterlony analyses employing reference BJ proteins kindly supplied by Dr. Leonhard Korngold (6).Antisera.--Anti-Bence Jones sera (anti-B J) were made in rabbits by immunization with the Sin. or S1. BJ protein in complete Freund's adjuvant, followed by intravenous administration of alum precipitates. The resulting antisera were absorbed in small increments with BJ protein and isolated 7S myeloma globulin of the opposite type, and finally with the fast fragment of normal human 7S "),-globulin prepared by published methods (18,19). In Ouchterlony analysis the absorbed antisera gave a single precipitin line with Cohn Fr. II, and this line fused completely with the lines given by several BJ proteins and myeloma globulins of the corresponding type and with the line given by L chains obtained from normal q/-giobulin (20). No precipifin lines were detected against a wide range of concentrations of the following antigens: BJ proteins and myeloma globulins of the opposite type, the fast fragment from normal "y-globulin, and the urinary "H chains" of Franklin's patient, Cr.
(21). s Anti-7S2 We are indebted to the following physicians for allowing us to study their patients: Dr. Scott N. Swisher, Dr. Stanley B. Troup, and Dr....