The Occurrence of Acid Mucopolysaccharides in Human Leukocytes and Urine 1

Acid mucopolysaccharides have been found in urine (1) but have not been consistently demon- strated in the blood of normal subjects.Bassiouni (2) recently isolated a substance from leukocytes which he noted to be similar to chondroitin sul- fate.In our laboratory leukocytes have been found to contain appreciable quantities of a sub- stance resembling chondroitin sulfate.A direct relationship between this substance and the simi- lar substance found in the urine has not been established. METHODSSubjects.Human subjects were used.These were hospital and laboratory personnel and patients who were unselected except for the omission of individuals with primary hematologic, metabolic, or rheumatic diseases.Preparation of leukocytes.Siliconed needles, syringes, and glassware were used throughout the preparation un- til the step in which the cells were deliberately disinte- grated.Venous blood (40 to 70 ml.)was drawn and distributed immediately in 10-ml.amounts to tubes con- taining 1.0 ml. of 5 per cent sodium citrate in a balanced salt solution.2The red blood cells were sedimented rapidly by mixing 2.5 ml. of 5 per cent dextran with the blood in each tube.3The supernatant leukocyte-platelet suspension in plasma was centrifuged at 85 g for 7 min- utes, and the sedimented leukocytes were washed gently one time in 11.0 ml. of physiologic saline.Following recentrifuging, they were then pooled in 20 ml. of saline, counted, made up to 25 ml.with saline in process of trans- ferring to a Raytheon Sonic Oscillator, and oscillated at 9 Kc. per second for 8 minutes.