Antibodies for denatured human H-ferritin stain only reticuloendothelial cells within the bone marrow

Br J Haematol. 1992 May;81(1):118-24. doi: 10.1111/j.1365-2141.1992.tb08183.x.

Abstract

Human H-ferritin homopolymer was denatured in sodium dodecyl sulphate and injected in mice to obtain antibodies for dissociated H-subunit. The antisera and Moabs obtained were specific for the denatured H-chain with no cross-reactivity with assembled ferritins in immunoblotting experiments. In contrast the Moabs for native recombinant H-ferritin are specific for the assembled ferritin molecules with weak cross-reactivity with the denatured H-subunits. The epitope recognized by one of the anti-denatured H-chain Moabs was mapped on the C-terminal helix of ferritin. The antibodies were used to study H-ferritin conformation in cells. In immunocytochemistry experiments the antibodies for denatured H-ferritin stained HeLa and K562 cells weakly, with a different intensity and pattern to those obtained with anti-native H-ferritin antibody. In human bone marrow smears the anti-denatured ferritin antibodies stained only reticuloendothelial cells, and did not recognize the H-ferritin rich immature erythroblasts. It is concluded that assembled and denatured H-ferritins are immunogenically distinct, and that erythroid and reticuloendothelial cells within the bone marrow contain H-ferritin in different conformations.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal*
  • Blotting, Western
  • Bone Marrow / chemistry
  • Bone Marrow / immunology
  • Bone Marrow Cells*
  • Electrophoresis, Polyacrylamide Gel
  • Erythroid Precursor Cells / chemistry
  • Erythroid Precursor Cells / cytology
  • Erythroid Precursor Cells / immunology
  • Ferritins / immunology*
  • Fluorescent Antibody Technique
  • HeLa Cells
  • Humans
  • Immunohistochemistry
  • Iron / metabolism
  • Mononuclear Phagocyte System / chemistry*
  • Mononuclear Phagocyte System / cytology*
  • Protein Denaturation
  • Staining and Labeling

Substances

  • Antibodies, Monoclonal
  • Ferritins
  • Iron