Structural, functional, and tissue distribution analysis of human transferrin receptor-2 by murine monoclonal antibodies and a polyclonal antiserum

Silvia Deaglio, Andrea Capobianco, Angelita Calì, Francesca Bellora, Federica Alberti, Luisella Righi, Anna Sapino, Clara Camaschella, Fabio Malavasi

Research output: Contribution to journalArticle

59 Citations (Scopus)

Abstract

Human transferrin receptor-2 (TFR-2) is a protein highly homologous to TFR-1/CD71 and is endowed with the ability to bind transferrin (TF) with low affinity. High levels of TFR-2 mRNA were found in the liver and in erythroid precursors. Mutations affecting the TFR-2 gene led to hemochromatosis type 3, a form of inherited iron overload. Several issues on distribution and function of the receptor were answered by raising a panel of 9 monoclonal antibodies specific for TFR-2 by immunizing mice with murine fibroblasts transfected with the human TFR-2 cDNA. A polyclonal antiserum was also produced in mice immunized with 3 peptides derived from the TFR-2 sequence, exploiting an innovative technique. The specificity of all the reagents produced was confirmed by reactivity with TFR-2+ target cells and simultaneous negativity with TFR-1+ cells. Western blot analyses showed a dominant chain of approximately 90 kDa in TFR-2 transfectants and HepG2 cell line. Analysis of distribution in normal tissues and in representative cell lines revealed that TFR-2 displays a restricted expression pattern - it is present at high levels in hepatocytes and in the epithelial cells of the small intestine, including the duodenal crypts. Exposure of human TFR-2+ cells to TF-bound iron is followed by a significant up-regulation and relocalization of membrane TFR-2. The tissue distribution pattern, the behavior following exposure to iron-loaded TF, and the features of the disease resulting from TFR-2 inactivation support the hypothesis that TFR-2 contributes to body iron sensing.

Original languageEnglish
Pages (from-to)3782-3789
Number of pages8
JournalBlood
Volume100
Issue number10
DOIs
Publication statusPublished - Nov 15 2002

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Transferrin Receptors
Tissue Distribution
Immune Sera
Monoclonal Antibodies
Tissue
Transferrin
Iron
Cells
human TFR2 protein
Cell Line
Aptitude
Iron Overload
Normal Distribution
Hep G2 Cells
Fibroblasts
Liver
Small Intestine
Hepatocytes
Up-Regulation
Complementary DNA

ASJC Scopus subject areas

  • Hematology

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Structural, functional, and tissue distribution analysis of human transferrin receptor-2 by murine monoclonal antibodies and a polyclonal antiserum. / Deaglio, Silvia; Capobianco, Andrea; Calì, Angelita; Bellora, Francesca; Alberti, Federica; Righi, Luisella; Sapino, Anna; Camaschella, Clara; Malavasi, Fabio.

In: Blood, Vol. 100, No. 10, 15.11.2002, p. 3782-3789.

Research output: Contribution to journalArticle

Deaglio, Silvia ; Capobianco, Andrea ; Calì, Angelita ; Bellora, Francesca ; Alberti, Federica ; Righi, Luisella ; Sapino, Anna ; Camaschella, Clara ; Malavasi, Fabio. / Structural, functional, and tissue distribution analysis of human transferrin receptor-2 by murine monoclonal antibodies and a polyclonal antiserum. In: Blood. 2002 ; Vol. 100, No. 10. pp. 3782-3789.
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