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. 1967 Mar 31;125(4):595–606. doi: 10.1084/jem.125.4.595

DEMONSTRATION BY IMMUNOFLUORESCENCE OF THE FIXATION OF PERFUSED ANTIBODY TO HUMAN COLLAGEN IN HUMAN KIDNEY

Sidney Rothbard 1, Robert F Watson 1
PMCID: PMC2138374  PMID: 4289739

Abstract

Rabbit serum containing antibody to human collagen, perfused through human infant kidneys obtained at autopsy, gives an immunofluorescent reaction with an antigen in the basement membranes of the glomeruli and the tubules. This reaction was shown to be specific by the absence of reaction with normal rabbit serum, antibody to carp collagen, or anti-human collagen serum absorbed with human collagen. Slight cross-reactions were found in the human kidneys with antibody to chicken or rat collagen. There was no evidence that a collagen-like protein in human serum or an antigen common to human erythrocytes and renal glomeruli enters into this immunofluorescent reaction. Perfusion of the kidneys with purified collagenase before the introduction of the antibody to human collagen altered the antigen so that antibody could not be demonstrated in the basement membranes by immunofluorescence. Testicular hyaluronidase used in the same way did not affect the immunofluorescent reaction. This method of perfusion provides another means for studying antigens in human organs by immunofluorescence. These observations in human kidneys extend our earlier findings in laboratory animals and indicate that an antigen, collagen, is also present in human renal glomerular and tubular basement membranes.

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Selected References

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  1. Bonting S. L., De Bruin H., Pollak V. E. QUANTITATIVE HISTOCHEMISTRY OF THE NEPHRON. VI. HYDROXYPROLINE IN THE HUMAN GLOMERULUS. J Clin Invest. 1961 Feb;40(2):177–180. doi: 10.1172/JCI104242. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. COONS A. H., KAPLAN M. H. Localization of antigen in tissue cells; improvements in a method for the detection of antigen by means of fluorescent antibody. J Exp Med. 1950 Jan 1;91(1):1–13. doi: 10.1084/jem.91.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. FARQUHAR M. G., VERNIER R. L., GOOD R. A. Studies on familial nephrosis. II. Glomerular changes observed with the electron microscope. Am J Pathol. 1957 Jul-Aug;33(4):791–817. [PMC free article] [PubMed] [Google Scholar]
  4. Frey J., Hoa N., Gras J., Henry J. C., Gudefin Y. Immunoelectrophoresis reveals collagen solubility in human serum. Science. 1965 Nov 5;150(3697):751–752. doi: 10.1126/science.150.3697.751. [DOI] [PubMed] [Google Scholar]
  5. GALLOP P. M., SEIFTER S., MEILMAN E. Studies on collagen. I. The partial purification, assay, and mode of activation of bacterial collagenase. J Biol Chem. 1957 Aug;227(2):891–906. [PubMed] [Google Scholar]
  6. GOODMAN M., GREENSPON S. A., KRAKOWER C. A. The antigenic composition of the various anatomic structures of the canine kidney. J Immunol. 1955 Aug;75(2):96–104. [PubMed] [Google Scholar]
  7. HAYWARD N. J. COLLAGENASE MACERATION IN RENAL MICRODISSECTION PRESERVATION OF NUCLEAR STAINING. Br J Exp Pathol. 1964 Feb;45:68–74. [PMC free article] [PubMed] [Google Scholar]
  8. KEISER H., LEROY E. C., UDENFRIEND S., SJOERDSMA A. COLLAGEN-LIKE PROTEIN IN HUMAN PLASMA. Science. 1963 Dec 27;142(3600):1678–1679. doi: 10.1126/science.142.3600.1678. [DOI] [PubMed] [Google Scholar]
  9. Kefalides N. A., Winzler R. J. The chemistry of glomerular basement membrane and its relation to collagen. Biochemistry. 1966 Feb;5(2):702–713. doi: 10.1021/bi00866a043. [DOI] [PubMed] [Google Scholar]
  10. LATTA H., MAUNSBACH A. B., MADDEN S. C. The centrolobular region of the renal glomerulus studied by electron microscopy. J Ultrastruct Res. 1960 Dec;4:455–472. doi: 10.1016/s0022-5320(60)80033-0. [DOI] [PubMed] [Google Scholar]
  11. MARKOWITZ A. S., LANGE C. F., Jr STREPTOCOCCAL RELATED GLOMERULONEPHRITIS. I. ISOLATION, IMMUNOCHEMISTRY AND COMPARATIVE CHEMISTRY OF SOLUBLE FRACTIONS FROM TYPE 12 NEPHRITOGENIC STREPTOCOCCI AND HUMAN GLOMERULI. J Immunol. 1964 Apr;92:565–575. [PubMed] [Google Scholar]
  12. MUELLER C. B., MASON A. D., Jr, STOUT D. G. Anatomy of the glomerulus. Am J Med. 1955 Feb;18(2):267–276. doi: 10.1016/0002-9343(55)90242-4. [DOI] [PubMed] [Google Scholar]
  13. PAK POY R. K. Electron microscopy of the mammalian renal glomerulus; the problems of intercapillary tissue and the capillary loop basement membrane. Am J Pathol. 1958 Sep-Oct;34(5):885–895. [PMC free article] [PubMed] [Google Scholar]
  14. PEASE D. C. Fine structures of the kidney seen by electron microscopy. J Histochem Cytochem. 1955 Jul;3(4):295–308. doi: 10.1177/3.4.295. [DOI] [PubMed] [Google Scholar]
  15. RINEHART J. F., FARQUHAR M. G., JUNG H. C., ABUL-HAJ S. The normal glomerulus and its basic reactions in disease. Am J Pathol. 1953 Jan-Feb;29(1):21–31. [PMC free article] [PubMed] [Google Scholar]
  16. ROTHBARD S., WATSON R. F. Antigenicity of rat collagen. Demonstration of antibody to rat collagen in the renal glomeruli of rats by fluorescence microscopy. J Exp Med. 1961 Jun 1;113:1041–1051. doi: 10.1084/jem.113.6.1041. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Rothbard S., Watson R. F. Immunologic relations among various animal collagens. J Exp Med. 1965 Sep 1;122(3):441–454. doi: 10.1084/jem.122.3.441. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. SEIFTER S., GALLOP P. M., KLEIN L., MEILMAN E. Studies on collagen. II. Properties of purified collagenase and its inhibition. J Biol Chem. 1959 Feb;234(2):285–293. [PubMed] [Google Scholar]
  19. Swift H. F., Wilson A. T., Lancefield R. C. TYPING GROUP A HEMOLYTIC STREPTOCOCCI BY M PRECIPITIN REACTIONS IN CAPILLARY PIPETTES. J Exp Med. 1943 Aug 1;78(2):127–133. doi: 10.1084/jem.78.2.127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. TAN E. M., KAPLAN M. H. IMMUNOLOGICAL RELATION OF BASEMENT MEMBRANE AND A SERUM BETA GLOBULIN IN THE MOUSE. DEMONSTRATION OF RENAL BASEMENT MEMBRANE ALTERATION IN MICE INJECTED WITH STREPTOLYSIN S. Immunology. 1963 Jul;6:331–344. [PMC free article] [PubMed] [Google Scholar]
  21. WATSON R. F., ROTHBARD S., VANAMEE P. The antigenicity of rat collagen. J Exp Med. 1954 Jun 1;99(6):535–550. doi: 10.1084/jem.99.6.535. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. YAMADA E. The fine structure of the renal glomerulus of the mouse. J Biophys Biochem Cytol. 1955 Nov 25;1(6):551–566. doi: 10.1083/jcb.1.6.551. [DOI] [PMC free article] [PubMed] [Google Scholar]

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