title: Monoclonal antibodies against the human sodium iodide symporter: utility for immunocytochemistry of thyroid cancer.
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) 1kappa (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 approximately 97 kDa, and several minor bands with MW of approximately 160 kDa, approximately 68 kDa, approximately 30 kDa and approximately 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 Hürthle 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.
authors.affiliation: Division of Endocrinology, Mayo Clinic and Mayo Graduate Medical School, Rochester, Minnesota, U S A.
authors.forename: M R
authors.initials: MR
authors.lastname: Castro
authors.forename: E R
authors.initials: ER
authors.lastname: Bergert
authors.forename: T G
authors.initials: TG
authors.lastname: Beito
authors.forename: P C
authors.initials: PC
authors.lastname: Roche
authors.forename: S C
authors.initials: SC
authors.lastname: Ziesmer
authors.forename: S M
authors.initials: SM
authors.lastname: Jhiang
authors.forename: J R
authors.initials: JR
authors.lastname: Goellner
authors.forename: J C
authors.initials: JC
authors.lastname: Morris
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification* / Q000627:therapeutic use; D000918:Antibody Specificity; D015153:Blotting, Western; D019556:COS Cells; D002352:Carrier Proteins / Q000032:analysis* / Q000276:immunology; D006801:Humans; D007074:Immunoglobulin G / Q000302:isolation & purification* / Q000627:therapeutic use; D007150:Immunohistochemistry; D037241:Mannose-Binding Lectins; D008565:Membrane Proteins / Q000032:analysis* / Q000276:immunology; D051379:Mice; D009363:Neoplasm Proteins / Q000032:analysis* / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000276:immunology; D027981:Symporters; D013964:Thyroid Neoplasms / Q000737:chemistry*; D000099209:Sodium-Iodide Symporters
chemical_list: D000911:Antibodies, Monoclonal; D002352:Carrier Proteins; D007074:Immunoglobulin G; D037241:Mannose-Binding Lectins; D008565:Membrane Proteins; D009363:Neoplasm Proteins; D011993:Recombinant Fusion Proteins; D027981:Symporters; D000099209:Sodium-Iodide Symporters