Regulation of major histocompatibility class ii gene expression in frtl-5 thyrocytes: Opposite effects of interferon and methimazole

Leonard D. Kohn, Minho Shong, Shinichi Taniguchi, Koichi Suzuki, Cesidio Giuliani, Giorgio Napolitano, Jun Saito, Motoyasu Saji, Bruno Fiorentino, Andreas M. Reimold, Dinah S. Singer, Leonard D. Kohn

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

Aberrant expression of major histocompatibility complex (MHC) class II antigens is associated with autoimmune thyroid disease; aberrant expression duplicating the autoimmune state can be induced by interferon-γ (IFNγ). We have studied IFNγ-induced human leukocyte antigen (HLA)-DRα gene expression in rat FRTL-5 thyroid cells to identify the elements and factors important for aberrant expression. Using an HLA-DRα 5'-flanking region construct from -176 to +45 bp coupled to the chloramphenicol acetyltransferase reporter gene, we show that there is no basal class II gene expression in FRTL-5 thyroid cells, that IFNγ can induce expression, and, as is the case for antigen-presenting cells from the immune system, that IFNγ-induced expression requires several highly conserved elements on the 5'-flanking region, which, from 5' to 3', are the S, X1, X2, and Y boxes. Methimazole (MMI), a drug used to treat patients with Graves' disease and experimental thyroiditis in rats or mice, can suppress the IFNγ-induced increase in HLA-DRα gene expression as a function of time and concentration; MMI simultaneously decreases IFNγ-induced endogenous antigen presentation by the cell. Using gel shift assays and the HLA-DRα 5'-flanking region from -176 or -137 to +45 bp as radiolabeled probes, we observed the formation of a major protein-DNA complex with extracts from FRTL-5 cells untreated with IFNγ, termed the basal or constitutive complex, and formation of an additional complex with a slightly faster mobility in extracts from cells treated with IFNγ. MMI treatment of cells prevents IFNγ from increasing the formation of this faster migrating complex. Formation of both complexes is specific, as evidenced in competition studies with unlabeled fragments between -137 and -38 bp from the start of transcription; nevertheless, they can be distinguished in such studies. Thus, high concentrations of double stranded oligonucleotides containing the sequence of the Y box, but not S, X1, or X2 box sequences, can prevent formation of the IFNγ-increased faster migrating complex, but not the basal complex. Both complexes involve multiple proteins and can be distinguished by differences in their protein composition. Thus, using specific antisera, we show that two cAMP response element-binding proteins, activating transcription factor-1 and/or -2, are dominant proteins in the upper or basal complex. The upper or basal complex also includes c-Fos, Fra-2, Ets-2, and Oct-1. A dominant protein that distinguishes the IFNγ-increased lower complex is CREB-binding protein (CBP), a coactivator of cAMP response element-binding proteins. We, therefore, show that aberrant expression of MHC class II in thyrocytes, induced by IFNγ, is associated with the induction or increased formation of a novel protein-DNA complex and that its formation as well as aberrant class II expression are suppressed by MMI, a drug used to treat human and experimental autoimmune thyroid disease. Its component proteins differ from those in a major, basal, or constitutive protein-DNA complex formed with the class II 5'-flanking region in cells that are not treated with IFNγ and that do not express the class II gene.

Original languageEnglish (US)
Pages (from-to)290-302
Number of pages13
JournalEndocrinology
Volume139
Issue number1
DOIs
StatePublished - 1998

Fingerprint

Methimazole
Histocompatibility
Interferons
Gene Expression
5' Flanking Region
HLA Antigens
Proteins
Cyclic AMP Response Element-Binding Protein
MHC Class II Genes
Thyroid Diseases
Major Histocompatibility Complex
Autoimmune Diseases
Thyroid Epithelial Cells
DNA
Activating Transcription Factor 1
Thyroid Gland
CREB-Binding Protein
Thyroiditis
Chloramphenicol O-Acetyltransferase
Graves Disease

ASJC Scopus subject areas

  • Endocrinology

Cite this

Kohn, L. D., Shong, M., Taniguchi, S., Suzuki, K., Giuliani, C., Napolitano, G., ... Kohn, L. D. (1998). Regulation of major histocompatibility class ii gene expression in frtl-5 thyrocytes: Opposite effects of interferon and methimazole. Endocrinology, 139(1), 290-302. https://doi.org/10.1210/endo.139.1.5658

Regulation of major histocompatibility class ii gene expression in frtl-5 thyrocytes : Opposite effects of interferon and methimazole. / Kohn, Leonard D.; Shong, Minho; Taniguchi, Shinichi; Suzuki, Koichi; Giuliani, Cesidio; Napolitano, Giorgio; Saito, Jun; Saji, Motoyasu; Fiorentino, Bruno; Reimold, Andreas M.; Singer, Dinah S.; Kohn, Leonard D.

In: Endocrinology, Vol. 139, No. 1, 1998, p. 290-302.

Research output: Contribution to journalArticle

Kohn, LD, Shong, M, Taniguchi, S, Suzuki, K, Giuliani, C, Napolitano, G, Saito, J, Saji, M, Fiorentino, B, Reimold, AM, Singer, DS & Kohn, LD 1998, 'Regulation of major histocompatibility class ii gene expression in frtl-5 thyrocytes: Opposite effects of interferon and methimazole', Endocrinology, vol. 139, no. 1, pp. 290-302. https://doi.org/10.1210/endo.139.1.5658
Kohn, Leonard D. ; Shong, Minho ; Taniguchi, Shinichi ; Suzuki, Koichi ; Giuliani, Cesidio ; Napolitano, Giorgio ; Saito, Jun ; Saji, Motoyasu ; Fiorentino, Bruno ; Reimold, Andreas M. ; Singer, Dinah S. ; Kohn, Leonard D. / Regulation of major histocompatibility class ii gene expression in frtl-5 thyrocytes : Opposite effects of interferon and methimazole. In: Endocrinology. 1998 ; Vol. 139, No. 1. pp. 290-302.
@article{b1d5832c65994c26bf31b27c2ef93c4f,
title = "Regulation of major histocompatibility class ii gene expression in frtl-5 thyrocytes: Opposite effects of interferon and methimazole",
abstract = "Aberrant expression of major histocompatibility complex (MHC) class II antigens is associated with autoimmune thyroid disease; aberrant expression duplicating the autoimmune state can be induced by interferon-γ (IFNγ). We have studied IFNγ-induced human leukocyte antigen (HLA)-DRα gene expression in rat FRTL-5 thyroid cells to identify the elements and factors important for aberrant expression. Using an HLA-DRα 5'-flanking region construct from -176 to +45 bp coupled to the chloramphenicol acetyltransferase reporter gene, we show that there is no basal class II gene expression in FRTL-5 thyroid cells, that IFNγ can induce expression, and, as is the case for antigen-presenting cells from the immune system, that IFNγ-induced expression requires several highly conserved elements on the 5'-flanking region, which, from 5' to 3', are the S, X1, X2, and Y boxes. Methimazole (MMI), a drug used to treat patients with Graves' disease and experimental thyroiditis in rats or mice, can suppress the IFNγ-induced increase in HLA-DRα gene expression as a function of time and concentration; MMI simultaneously decreases IFNγ-induced endogenous antigen presentation by the cell. Using gel shift assays and the HLA-DRα 5'-flanking region from -176 or -137 to +45 bp as radiolabeled probes, we observed the formation of a major protein-DNA complex with extracts from FRTL-5 cells untreated with IFNγ, termed the basal or constitutive complex, and formation of an additional complex with a slightly faster mobility in extracts from cells treated with IFNγ. MMI treatment of cells prevents IFNγ from increasing the formation of this faster migrating complex. Formation of both complexes is specific, as evidenced in competition studies with unlabeled fragments between -137 and -38 bp from the start of transcription; nevertheless, they can be distinguished in such studies. Thus, high concentrations of double stranded oligonucleotides containing the sequence of the Y box, but not S, X1, or X2 box sequences, can prevent formation of the IFNγ-increased faster migrating complex, but not the basal complex. Both complexes involve multiple proteins and can be distinguished by differences in their protein composition. Thus, using specific antisera, we show that two cAMP response element-binding proteins, activating transcription factor-1 and/or -2, are dominant proteins in the upper or basal complex. The upper or basal complex also includes c-Fos, Fra-2, Ets-2, and Oct-1. A dominant protein that distinguishes the IFNγ-increased lower complex is CREB-binding protein (CBP), a coactivator of cAMP response element-binding proteins. We, therefore, show that aberrant expression of MHC class II in thyrocytes, induced by IFNγ, is associated with the induction or increased formation of a novel protein-DNA complex and that its formation as well as aberrant class II expression are suppressed by MMI, a drug used to treat human and experimental autoimmune thyroid disease. Its component proteins differ from those in a major, basal, or constitutive protein-DNA complex formed with the class II 5'-flanking region in cells that are not treated with IFNγ and that do not express the class II gene.",
author = "Kohn, {Leonard D.} and Minho Shong and Shinichi Taniguchi and Koichi Suzuki and Cesidio Giuliani and Giorgio Napolitano and Jun Saito and Motoyasu Saji and Bruno Fiorentino and Reimold, {Andreas M.} and Singer, {Dinah S.} and Kohn, {Leonard D.}",
year = "1998",
doi = "10.1210/endo.139.1.5658",
language = "English (US)",
volume = "139",
pages = "290--302",
journal = "Endocrinology",
issn = "0013-7227",
publisher = "The Endocrine Society",
number = "1",

}

TY - JOUR

T1 - Regulation of major histocompatibility class ii gene expression in frtl-5 thyrocytes

T2 - Opposite effects of interferon and methimazole

AU - Kohn, Leonard D.

AU - Shong, Minho

AU - Taniguchi, Shinichi

AU - Suzuki, Koichi

AU - Giuliani, Cesidio

AU - Napolitano, Giorgio

AU - Saito, Jun

AU - Saji, Motoyasu

AU - Fiorentino, Bruno

AU - Reimold, Andreas M.

AU - Singer, Dinah S.

AU - Kohn, Leonard D.

PY - 1998

Y1 - 1998

N2 - Aberrant expression of major histocompatibility complex (MHC) class II antigens is associated with autoimmune thyroid disease; aberrant expression duplicating the autoimmune state can be induced by interferon-γ (IFNγ). We have studied IFNγ-induced human leukocyte antigen (HLA)-DRα gene expression in rat FRTL-5 thyroid cells to identify the elements and factors important for aberrant expression. Using an HLA-DRα 5'-flanking region construct from -176 to +45 bp coupled to the chloramphenicol acetyltransferase reporter gene, we show that there is no basal class II gene expression in FRTL-5 thyroid cells, that IFNγ can induce expression, and, as is the case for antigen-presenting cells from the immune system, that IFNγ-induced expression requires several highly conserved elements on the 5'-flanking region, which, from 5' to 3', are the S, X1, X2, and Y boxes. Methimazole (MMI), a drug used to treat patients with Graves' disease and experimental thyroiditis in rats or mice, can suppress the IFNγ-induced increase in HLA-DRα gene expression as a function of time and concentration; MMI simultaneously decreases IFNγ-induced endogenous antigen presentation by the cell. Using gel shift assays and the HLA-DRα 5'-flanking region from -176 or -137 to +45 bp as radiolabeled probes, we observed the formation of a major protein-DNA complex with extracts from FRTL-5 cells untreated with IFNγ, termed the basal or constitutive complex, and formation of an additional complex with a slightly faster mobility in extracts from cells treated with IFNγ. MMI treatment of cells prevents IFNγ from increasing the formation of this faster migrating complex. Formation of both complexes is specific, as evidenced in competition studies with unlabeled fragments between -137 and -38 bp from the start of transcription; nevertheless, they can be distinguished in such studies. Thus, high concentrations of double stranded oligonucleotides containing the sequence of the Y box, but not S, X1, or X2 box sequences, can prevent formation of the IFNγ-increased faster migrating complex, but not the basal complex. Both complexes involve multiple proteins and can be distinguished by differences in their protein composition. Thus, using specific antisera, we show that two cAMP response element-binding proteins, activating transcription factor-1 and/or -2, are dominant proteins in the upper or basal complex. The upper or basal complex also includes c-Fos, Fra-2, Ets-2, and Oct-1. A dominant protein that distinguishes the IFNγ-increased lower complex is CREB-binding protein (CBP), a coactivator of cAMP response element-binding proteins. We, therefore, show that aberrant expression of MHC class II in thyrocytes, induced by IFNγ, is associated with the induction or increased formation of a novel protein-DNA complex and that its formation as well as aberrant class II expression are suppressed by MMI, a drug used to treat human and experimental autoimmune thyroid disease. Its component proteins differ from those in a major, basal, or constitutive protein-DNA complex formed with the class II 5'-flanking region in cells that are not treated with IFNγ and that do not express the class II gene.

AB - Aberrant expression of major histocompatibility complex (MHC) class II antigens is associated with autoimmune thyroid disease; aberrant expression duplicating the autoimmune state can be induced by interferon-γ (IFNγ). We have studied IFNγ-induced human leukocyte antigen (HLA)-DRα gene expression in rat FRTL-5 thyroid cells to identify the elements and factors important for aberrant expression. Using an HLA-DRα 5'-flanking region construct from -176 to +45 bp coupled to the chloramphenicol acetyltransferase reporter gene, we show that there is no basal class II gene expression in FRTL-5 thyroid cells, that IFNγ can induce expression, and, as is the case for antigen-presenting cells from the immune system, that IFNγ-induced expression requires several highly conserved elements on the 5'-flanking region, which, from 5' to 3', are the S, X1, X2, and Y boxes. Methimazole (MMI), a drug used to treat patients with Graves' disease and experimental thyroiditis in rats or mice, can suppress the IFNγ-induced increase in HLA-DRα gene expression as a function of time and concentration; MMI simultaneously decreases IFNγ-induced endogenous antigen presentation by the cell. Using gel shift assays and the HLA-DRα 5'-flanking region from -176 or -137 to +45 bp as radiolabeled probes, we observed the formation of a major protein-DNA complex with extracts from FRTL-5 cells untreated with IFNγ, termed the basal or constitutive complex, and formation of an additional complex with a slightly faster mobility in extracts from cells treated with IFNγ. MMI treatment of cells prevents IFNγ from increasing the formation of this faster migrating complex. Formation of both complexes is specific, as evidenced in competition studies with unlabeled fragments between -137 and -38 bp from the start of transcription; nevertheless, they can be distinguished in such studies. Thus, high concentrations of double stranded oligonucleotides containing the sequence of the Y box, but not S, X1, or X2 box sequences, can prevent formation of the IFNγ-increased faster migrating complex, but not the basal complex. Both complexes involve multiple proteins and can be distinguished by differences in their protein composition. Thus, using specific antisera, we show that two cAMP response element-binding proteins, activating transcription factor-1 and/or -2, are dominant proteins in the upper or basal complex. The upper or basal complex also includes c-Fos, Fra-2, Ets-2, and Oct-1. A dominant protein that distinguishes the IFNγ-increased lower complex is CREB-binding protein (CBP), a coactivator of cAMP response element-binding proteins. We, therefore, show that aberrant expression of MHC class II in thyrocytes, induced by IFNγ, is associated with the induction or increased formation of a novel protein-DNA complex and that its formation as well as aberrant class II expression are suppressed by MMI, a drug used to treat human and experimental autoimmune thyroid disease. Its component proteins differ from those in a major, basal, or constitutive protein-DNA complex formed with the class II 5'-flanking region in cells that are not treated with IFNγ and that do not express the class II gene.

UR - http://www.scopus.com/inward/record.url?scp=85011463316&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85011463316&partnerID=8YFLogxK

U2 - 10.1210/endo.139.1.5658

DO - 10.1210/endo.139.1.5658

M3 - Article

C2 - 9421427

AN - SCOPUS:85011463316

VL - 139

SP - 290

EP - 302

JO - Endocrinology

JF - Endocrinology

SN - 0013-7227

IS - 1

ER -