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The chemical mediation of delayed hypersensitivity skin reaction. II. Characterization of a macrophage-chemotactic factor from bovine gamma-globulin-induced skin reaction in guinea pigs.

Macrophage-chemotactic factors were extracted from delayed hypersensitivity skin lesions induced by bovine gamma-globulin in guinea pigs. The most active factor, MCFS--1, was highly purified and found to be a heat-labile protein with a molecular weight of 150,000 and to possess in vivo as well as in vitro activity. This factor was homogeneous during polyacrylamide gel electrophoresis, and the chemotactic activity was associated exclusively with this band. Further characterization revealed that its isoelectric point was 6.7 to 6.9 and made a single arc in the beta-globulin region with rabbit antiserums against guinea pig serum on immunoelectrophoresis. This factor seemed to be antigenically different from immunoglobulin G (IgG) by immunodiffusion and immunoadsorption. On the other hand, the chemotactic activity of MCFS-2 was adsorbed by neither anti-IgG nor anti MCFS-1 and that of euglobulin fraction was partially adsorbed by anti-IgG. These indicate the presence of at least three types of antigenically different chemotactic factors for macrophages in the extracts of delayed hypersensitivity skin lesions.

Animals

The main neutral protease of rat skin is a mast cell enzyme. Immunohistochemical localization of the enzyme in rat skin with the peroxidase-antiperoxidase (PAP) complex method.

The highly sensitive PAP immunoperoxidase method was used to localize the main neutral protease of rat skin. The use of the neutral detergent, Triton X-100, in the reagent and washing solutions was observed to effectively decrease the nonspecific staining. The specific staining was localized to the mast cell granules.

Animals

[Hydroxylation of skin collagen in relation to age: analyses of the human skin].

The results of human skin analysis are demonstrated. It has been pointed out, that the content of hydroxyproline and hydroxylysine, in addition to the usual variation as the other aminoacids varies extensively. The extent of hydroxylation was estimated from the relation of OH-aminoacids to (not hydroxylated) each other. The determined variation of the results is most probably due to a different collagen hydroxylation within the biological dispersion. An age-related degree of hydroxylation could not be detected.

Adult

The chemical mediation of delayed hypersensitivity skin reactions. I. Purification of a macrophage-chemotactic factor from bovien gamma-globulin-induced skin reactions in guinea pigs.

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, two 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 found to be a protein that was free of nucleic acid. Gel filtration showed that its molecular weight was similar to that of IgG. 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 macrophage reaction in the delayed hypersensitivity reactions. Am J Pathol 87:359-374, 1977).

Animals