Monoclonal antibodies against the human sodium iodide symporter: Utility for immunocytochemistry of thyroid cancer

M. R. Castro, E. R. Bergert, T. G. Beito, P. C. Roche, S. C. Ziesmer, S. M. Jhiang, J. R. Goellner, J. C. Morris

Research output: Contribution to journalArticle

65 Scopus citations

Abstract

The recent cloning of the thyroidal protein that is responsible for iodide transport, the sodium iodide symporter (hNIS), has made possible studies designed to characterize its structure, function and expression in thyroidal tissues. Using a mannose binding protein (MBP)-hNIS fusion protein as antigen, we have developed mouse monoclonal antibodies against hNIS to utilize as tools in such studies. Twenty-four clones were initially recovered which recognized the MBP-hNIS fusion protein, but only two of them were specific for hNIS while the others recognized MBP alone. Both antibodies were found to be immunoglobulin G (IgG) 1κ (kappa). The specificity of antibodies was tested by Western blotting using membranes prepared from COS-7 cells transiently transfected with the pcDNA3 plasmid containing the full-length hNIS cDNA, or cells transfected with the pcDNA3 vector. A major band with a molecular weight (MW) of ~ 97 kDa, and several minor bands with MW of ~160 kDa, ~68 kDa, ~30 kDa and ~15 kDa, were detected specifically in the hNIS- transfected cells. After enzymatic deglycosylation, the major band was present at 68 kDa, as expected based upon the amino acid sequence of hNIS. Immunohistochemistry was performed with several different types of thyroid tissue and non-thyroidal tissues, using the monoclonal antibodies. Strong immunostaining was observed in Graves' tissue, with intermediate staining in papillary and follicular thyroid cancers and an absence of staining in Hurthle cell cancer. The staining was specific for the follicular epithelium and was concentrated in the basolateral portion of the cell membrane. These monoclonal hNIS antibodies should prove useful in the characterization of NIS expression in benign and malignant thyroid tissue and in studies characterizing its structure and function.

Original languageEnglish (US)
Pages (from-to)495-504
Number of pages10
JournalJournal of Endocrinology
Volume163
Issue number3
DOIs
StatePublished - Jan 1 1999

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Endocrinology

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    Castro, M. R., Bergert, E. R., Beito, T. G., Roche, P. C., Ziesmer, S. C., Jhiang, S. M., Goellner, J. R., & Morris, J. C. (1999). Monoclonal antibodies against the human sodium iodide symporter: Utility for immunocytochemistry of thyroid cancer. Journal of Endocrinology, 163(3), 495-504. https://doi.org/10.1677/joe.0.1630495