CHEMICAL, ULTRASTRUCTURAL AND CLINICAL STUDIES ON A PYROGEN ISOLATED FROM LEUCOCYTES.
Final comprehensive rept., 1 Jun 63-31 Aug 66,
ALBANY MEDICAL COLL N Y
Pagination or Media Count:
Standardized methods of producing leucocytic pyrogen, using either rabbit or human leucocytes, were developed. Studies of the mode of production of pyrogen by the leucocytes using the inhibitors actinomycin D, chloromycetin, mitomycin C and puromycin confirmed involvement of some synthetic process since puromycin was found to inhibit pyrogen production, though the other inhibitors did not. Pyrogen production was also studied through measurement of incorporation of amino acids, both unlabeled and C14 labeled. Methods of isolating and purifying leucocytic pyrogen were refined. These procedures include chromatography using DEAE and phosphorylated cellulose columns, monitored by electrophoresis and bioassay. Standardization of assay procedures for rabbit leucocytic pyrogen in rabbits was achieved. A procedure whereby human leucocytic pyrogen can be assayed in rabbits was developed. Characterization of leucocytic pyrogen was approached by studying its structure, activity, possible relation to virus, and the chemical constitution of various structures related to the purified pyrogen. The active site of the pyrogen molecule was found to be related in some way to the osmiophilic area morphologically demonstrated by electron microscopy. Author