Isolation and Characterization of Glycosaminoglycans from Pulmonary Secretions of Patients with Alveolar Proteinosis

Abstract

Alveolar proteinosis is a chronic pulmonary disease of unknown pathogenesis in which the alveoli and terminal bronchioles of the lung are filled with insoluble, periodic-Schiff positive amorphous material. This material can be removed from the lung by therapeutic bronchoalveolar lavage. This chapter describes an experiment to0 study the isolation and characterization of glycosaminoglycans from the pulmonary secretions of patients with alveolar proteinosis. As much as 10–50 g of dry material was obtained from the insoluble lavage sediment and 2–10 g from the soluble supernatant fraction from the lungs of one patient, depending on the state of the disease. During the isolation and purification of phospholipase A 2 from these soluble pulmonary secretions, it was observed that certain material remained bound to the DEAE-cellulose column, even after elution with 1 M NaCl solution but could be eluted with dil. HCl. Cellulose acetate electrophoresis in three different electrolyte systems, followed by staining with Alcian Blue revealed that hyaluronic acid was the only detectable glycosaminoglycan present. In all the three systems, the compound migrated with the standard of hyaluronic acid and after digestion with testicular hyaluronidase, no spots were detected.