CHEMICAL MEDIATION OF DELAYED-HYPERSENSITIVITY SKIN REACTIONS .1. PURIFICATION OF A MACROPHAGE-CHEMOTACTIC FACTOR FROM BOVINE GAMMA-GLOBULIN-INDUCED SKIN REACTIONS IN GUINEA-PIGS

  • 1 January 1977
    • journal article
    • research article
    • Vol. 87  (2) , 359-374
Abstract
A macrophage-chemotactic factor (MCFS) was extracted in the pseudoglobulin fraction from delayed hypersensitivity skin lesions induced by bovine .gamma.-globulin in guinea pigs. Its chemotactic activity was estimated by a modification of Boyden''s method using Nuclepore filter. After chromatography of the protein fraction using Sephadex G-50 and DEAE-cellulose, in that order, 2 chemotactic fractions were obtained. The chemotactic factor with stronger activity (MCFS-1) was further highly purified (488-fold) by chromatography on CM-Sephadex. This factor migrated in a single band on acrylamide disc gel electrophoresis and was a nucleic acid-free protein. Gel filtration showed that its MW was similar to that of Ig[immunoglobulin]G. Its chemotactic activity was heat labile. Intradermal injection of this factor into normal guinea pigs induced a pronounced mononuclear cell emigration from venules. These findings are pertinent to understanding the macrophage reaction in the delayed hypersensitivity reactions.