mRNA for genes associated with antigen presentation are expressed by human middle meatal epithelial cells in culture

Andrew P Lane, Bahman Saatian, Xiao Ying Yu, Ernst W Spannhake

Research output: Contribution to journalArticle

Abstract

Objectives/Hypothesis: Although the mechanisms underlying the initiation and maintenance of inflammation in chronic rhinosinusitis are poorly understood, the activation of memory T cells within the nasal mucosa is thought to play an important role. T-cell activation requires specialized antigen processing and presentation of antigen by immunocompetent cells in the context of cell surface immune molecules. The purpose of this study was to investigate the expression of such molecules by human sinonasal epithelial cells grown in culture at the air-liquid interface (ALI). Methods: Middle meatal epithelium was obtained from six patients undergoing endoscopic sinus surgery. Dissociated epithelial cells were grown to confluence in serum-free, defined medium and transferred to filter inserts for culture at the ALI. Cells were harvested at 2 and 21 days of growth at the ALI and processed for real-time polymerase chain reaction (PCR). The presence and relative abundance of constitutively expressed mRNA for human leukocyte antigen (HLA)-B, HLA-DR, B7 to 1, B7 to 2, B7-H2, B7-H3, and cathepsin D were assessed. Results: After 2 days at the ALI, middle meatal epithelial cells demonstrated expression of genes for each of the antigen processing associated genes tested. The expression of HLA-B and HLA-DR increased significantly with cellular maturation at the ALI. Expression of HLA-DR and B7 to 1 increased with cytokine stimulation. Conclusions: Primary human epithelial cells obtained from the middle meatus express genes associated with antigen presentation function. The pattern of gene expression is modulated by cytokine stimulation and changes as the cells differentiate at the ALI. These findings suggest that mature middle meatal epithelial cells have the cellular machinery to interact with T cells and therefore may be direct participants in the modulation of T-cell activity in chronic sinusitis.

Original languageEnglish (US)
Pages (from-to)1827-1832
Number of pages6
JournalLaryngoscope
Volume114
Issue number10 I
DOIs
StatePublished - Oct 2004

Fingerprint

Antigen Presentation
HLA Antigens
Cell Culture Techniques
Epithelial Cells
Air
Messenger RNA
Genes
T-Lymphocytes
Cytokines
Gene Expression
Cathepsin D
Nasal Mucosa
Sinusitis
Serum-Free Culture Media
Real-Time Polymerase Chain Reaction
Epithelium
Maintenance
Inflammation
Antigens
Growth

Keywords

  • Antigen presentation
  • Cathepsin D
  • Co-stimulatory molecules
  • Epithelial cell culture
  • MHC molecules
  • Sinonasal epithelium

ASJC Scopus subject areas

  • Otorhinolaryngology

Cite this

mRNA for genes associated with antigen presentation are expressed by human middle meatal epithelial cells in culture. / Lane, Andrew P; Saatian, Bahman; Yu, Xiao Ying; Spannhake, Ernst W.

In: Laryngoscope, Vol. 114, No. 10 I, 10.2004, p. 1827-1832.

Research output: Contribution to journalArticle

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title = "mRNA for genes associated with antigen presentation are expressed by human middle meatal epithelial cells in culture",
abstract = "Objectives/Hypothesis: Although the mechanisms underlying the initiation and maintenance of inflammation in chronic rhinosinusitis are poorly understood, the activation of memory T cells within the nasal mucosa is thought to play an important role. T-cell activation requires specialized antigen processing and presentation of antigen by immunocompetent cells in the context of cell surface immune molecules. The purpose of this study was to investigate the expression of such molecules by human sinonasal epithelial cells grown in culture at the air-liquid interface (ALI). Methods: Middle meatal epithelium was obtained from six patients undergoing endoscopic sinus surgery. Dissociated epithelial cells were grown to confluence in serum-free, defined medium and transferred to filter inserts for culture at the ALI. Cells were harvested at 2 and 21 days of growth at the ALI and processed for real-time polymerase chain reaction (PCR). The presence and relative abundance of constitutively expressed mRNA for human leukocyte antigen (HLA)-B, HLA-DR, B7 to 1, B7 to 2, B7-H2, B7-H3, and cathepsin D were assessed. Results: After 2 days at the ALI, middle meatal epithelial cells demonstrated expression of genes for each of the antigen processing associated genes tested. The expression of HLA-B and HLA-DR increased significantly with cellular maturation at the ALI. Expression of HLA-DR and B7 to 1 increased with cytokine stimulation. Conclusions: Primary human epithelial cells obtained from the middle meatus express genes associated with antigen presentation function. The pattern of gene expression is modulated by cytokine stimulation and changes as the cells differentiate at the ALI. These findings suggest that mature middle meatal epithelial cells have the cellular machinery to interact with T cells and therefore may be direct participants in the modulation of T-cell activity in chronic sinusitis.",
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AU - Lane, Andrew P

AU - Saatian, Bahman

AU - Yu, Xiao Ying

AU - Spannhake, Ernst W

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N2 - Objectives/Hypothesis: Although the mechanisms underlying the initiation and maintenance of inflammation in chronic rhinosinusitis are poorly understood, the activation of memory T cells within the nasal mucosa is thought to play an important role. T-cell activation requires specialized antigen processing and presentation of antigen by immunocompetent cells in the context of cell surface immune molecules. The purpose of this study was to investigate the expression of such molecules by human sinonasal epithelial cells grown in culture at the air-liquid interface (ALI). Methods: Middle meatal epithelium was obtained from six patients undergoing endoscopic sinus surgery. Dissociated epithelial cells were grown to confluence in serum-free, defined medium and transferred to filter inserts for culture at the ALI. Cells were harvested at 2 and 21 days of growth at the ALI and processed for real-time polymerase chain reaction (PCR). The presence and relative abundance of constitutively expressed mRNA for human leukocyte antigen (HLA)-B, HLA-DR, B7 to 1, B7 to 2, B7-H2, B7-H3, and cathepsin D were assessed. Results: After 2 days at the ALI, middle meatal epithelial cells demonstrated expression of genes for each of the antigen processing associated genes tested. The expression of HLA-B and HLA-DR increased significantly with cellular maturation at the ALI. Expression of HLA-DR and B7 to 1 increased with cytokine stimulation. Conclusions: Primary human epithelial cells obtained from the middle meatus express genes associated with antigen presentation function. The pattern of gene expression is modulated by cytokine stimulation and changes as the cells differentiate at the ALI. These findings suggest that mature middle meatal epithelial cells have the cellular machinery to interact with T cells and therefore may be direct participants in the modulation of T-cell activity in chronic sinusitis.

AB - Objectives/Hypothesis: Although the mechanisms underlying the initiation and maintenance of inflammation in chronic rhinosinusitis are poorly understood, the activation of memory T cells within the nasal mucosa is thought to play an important role. T-cell activation requires specialized antigen processing and presentation of antigen by immunocompetent cells in the context of cell surface immune molecules. The purpose of this study was to investigate the expression of such molecules by human sinonasal epithelial cells grown in culture at the air-liquid interface (ALI). Methods: Middle meatal epithelium was obtained from six patients undergoing endoscopic sinus surgery. Dissociated epithelial cells were grown to confluence in serum-free, defined medium and transferred to filter inserts for culture at the ALI. Cells were harvested at 2 and 21 days of growth at the ALI and processed for real-time polymerase chain reaction (PCR). The presence and relative abundance of constitutively expressed mRNA for human leukocyte antigen (HLA)-B, HLA-DR, B7 to 1, B7 to 2, B7-H2, B7-H3, and cathepsin D were assessed. Results: After 2 days at the ALI, middle meatal epithelial cells demonstrated expression of genes for each of the antigen processing associated genes tested. The expression of HLA-B and HLA-DR increased significantly with cellular maturation at the ALI. Expression of HLA-DR and B7 to 1 increased with cytokine stimulation. Conclusions: Primary human epithelial cells obtained from the middle meatus express genes associated with antigen presentation function. The pattern of gene expression is modulated by cytokine stimulation and changes as the cells differentiate at the ALI. These findings suggest that mature middle meatal epithelial cells have the cellular machinery to interact with T cells and therefore may be direct participants in the modulation of T-cell activity in chronic sinusitis.

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KW - Cathepsin D

KW - Co-stimulatory molecules

KW - Epithelial cell culture

KW - MHC molecules

KW - Sinonasal epithelium

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