Demonstration by Immunofluorescence of the Fixation of Perfused Antibody to Human Collagen in Human Kidney

Abstract

Immunofluorescence has demonstrated that antibody prepared in rabbits against acid-soluble collagen from rat, mouse, guinea pig, or chicken, injected into the circulation of the homologous species, becomes fixed in the basement membranes of the renal glomeruli and tubules (1, 2). In vitro and in vivo experiments have indicated that the antibody is directed specifically toward collagen and that it exhibits species specificity (1-3). These observations indicate that some form of collagen is present in the basement membranes even though most observers have not found collagen fibrils in normal mammalian renal glomeruli (4--10). The high proportion of hydroxyproline present on amino acid analysis of renal glomerular basement membranes of the dog (11, 12) and of man (13-15) also strongly suggests the presence of collagen.It therefore seemed desirable to extend our observations in animals to human renal glomeruli. The injection of collagen antibody into the circulation of the animals, a procedure not feasible in man, gave better defined and more intense fluorescence than the method in which frozen kidney sections were treated with the anticollagen serum (1). We therefore attempted to approximate the injection method by perfusing the collagen antibody through the renal artery of kidneys of human infants obtained at autopsy. This paper presents the results of immunofluorescence by this method and shows that antibody to human collagen becomes fixed to its antigen, collagen, in the human renal glomerular and tubular basement membranes. Materials and MethodsPurified acid-soluble collagen prepared from human skin, rat tail tendons, chicken leg tendons, or carp swim bladder, and rabbit antisera to these collagens were prepared by methods previously reported (2, 3). The technique of complement fixation for determining the titer of

Isolation and characterization of antigenic components of rat glomerular basement membrane

“…10). Antibody to tendon collagen was also found to be non-nephrotoxic in rats despite their localization on rat GBM (Table V); this is in agreement with previous reports (12)(13)(14)(15). Tt can be noted that anti-CSColl did not localize on kidney, suggesting that peptide isolated from the collagenase digest of tendon collagen did not contain any common antigenic determinants with GBM or that the binding sites on intact membrane were unavailable.…”

Immunological characterization of some basement membrane antigens.

“…Markowitz and Lange (1964) found an immunological cross-reaction between human glomeruli and cell membranes of nephritogenic strains of streptococci, and obtained low molecular weight glycoproteins from both sources. Rothbard and Watson (1967) obtained an immunofluorescent reaction in the glomerular basement membrane of human kidneys which had been perfused with antihuman-collagen serum. Collagenase pre-treatment prevented this reaction; it was concluded that collagen in human basement membrane was the relevant antigen.…”

Precipitating and Complement-Fixing Antibodies to Collagen with Species and Collagen Subunit Specificity

“…These studies have shown that the collagens of several species differ considerably in their antigenic character and presumably therefore in primary structure. These studies have also shown the presence of a material reacting with collagen antibodies in the renal glomerular basement membrane of the rat and man (9). A crucial feature of the antigenicity of tropocollagen, the major constituent of all extractable or "soluble" collagens, was the demonstration by Schmitt et al that the antigenic determinants of tropocollagen reside largely in regions of the molecule essential to the cross-linking of collagen monomers into the various polymers which can be formed (12).…”

“…In addition, and perhaps more significantly, free ACTH is, of course, steroidogenic and the steroids thus elicited result in depression of antibody synthesis (9). This problem has been partially circumvented by the use of mepirapone, a chemical agent which blocks 11-/I hydroxylation of the steroid nucleus (7).…”