Orosomucoid content of pleural and peritoneal effusions.

22 nonneoplastic, noninflammatory effusions (cirrhosis and congestive heart failure), 12 non-neoplastic inflammatory effusions (tuberculosis, lupus erythematosus, rheumatoid arthritis, and idiopathic pleuropericarditis), and 58 neoplastic effusions (cancer of lung, breast, ovary, and pancreas, and lymphoma) were analyzed by radial immunodiffusion for orosomucoid concentration. The average concentration +/-SE was 35+/-4, 65+/-17, and 130+/-13 mg/100 ml in the three types of effusion, respectively. By gel filtration and ion exchange chromatography, orosomucoid was isolated from 12 nonmalignant and 14 malignant fluids. The orosomucoid preparations reacted as single components in acrylamide gel electrophoresis at pH 9.0, and in immunodiffusion and immunoelectrophoresis against antisera to human serum and to human plasma orosomucoid. In radial immunodiffusion, the slope of the line relating concentration to the square of the diameter of the precipitate area was identical for orosomucoid isolated from normal human plasma and from nonneoplastic effusions, but was subnormal for orosomucoid isolated from neoplastic fluids. All orosomucoid preparations had normal amino acid composition. Orosomucoid from the nonmalignant effusions had normal carbohydrate content. 11 of 14 samples of orosomucoid isolated from neoplastic fluids had abnormalities in carbohydrate composition, consisting of subnormal content of sialic acid (11 of 14), hexose (10 of 14), and hexosamine (3 of 14), and abnormally high content of hexosamine (4 of 14). Discriminant analysis showed that concentration of orosomucoid distinguished between neoplastic and nonneoplastic noninflammatory effusions more effectively than concentration of total protein, albumin, alpha(1), alpha(2), beta, or gamma-globulin.

Isolation of a novel glycoprotein from the urine of a patient with chronic myelocytic leukemia.

Patient B. J. with chronic myelocytic leukemia excreted 0.5-1.1 g protein per day in the urine. Gel filtration on Sephadex G-75 showed about one-third of this protein to be in molecular weight range 20,000-40,000 (fraction BJC). BJC, prepared from 9 liters of urine by gel filtration, was chromatographed on carboxymethylcellulose. Two proteins were eluted from the resin in pure form (as shown by zone and immunoelectrophoresis) in yields representing 8 and 3 mg/liter of urine: BJC1 and BJC2. Their amino acid compositions were identical. BJC1 contained 61% carbohydrate (33% hexose, 11% sialic acid, 13% glucosamine, 5% galactosamine). BJC2 contained one-fourth to one-half as much of each carbohydrate. Molecular weight of BJC1 was estimated at 29,000 by gel filtration. Neither glycoprotein reacted with rabbit antiserum to normal human serum.Antiserum to BJC1 was made in the rabbit. Immunoelectrophoresis with this antiserum showed a faint precipitin line, corresponding in mobility to BJC1, in normal human plasma, and a stronger line in most leukemic plasmas. By immunodiffusion, BJC1 was not detectable in normal human urine, but a positive reaction occurred in the following conditions: leukemia, 64-72%; other types of disseminated neoplastic disease, 36-78%; regional ileitis, 45%; ulcerative colitis, 38%; tuberculosis, 33%; during the 1st wk after major surgery, 33%.BJC2 was found in the urine by immunoelectrophoresis in 10% of patients with neoplastic disease and was not observed in urine of other patients or in human plasma. Amino acid composition, carbohydrate content, and antigenic specificity indicate BJC1 is a previously unrecognized member of the system of normal human plasma glycoproteins. Like certain other glycoproteins, its plasma concentration frequently increases in patients with neoplastic disease, chronic inflammatory disease, or tuberculosis and after surgery. Because molecular weight is 29,000, increased plasma concentration readily causes its appearance in the urine.