Abstract
Background: Eosinophils are now known to produce a variety of proinflammatory cytokines, although the molecular factors that regulate their production are poorly understood. The expression of almost all of the cytokines produced by eosinophils, including the proallergic cytokine IL-4, is now known to be regulated at the level of transcription by members of the nuclear factor of activated T cells (NFAT) family of transcription factors. Objective: We sought to characterize the expression of different NFAT proteins in resting and activated eosinophils. Methods: Nuclear and whole cell extracts were obtained from both peripheral blood eosinophils and those obtained from bronchoalveolar lavage fluid of asthmatic subjects after endobronchial allergen challenge. NFAT expression was determined by using immunoprecipitation and Western blot analysis, DNA-binding assays, and RT-PCR analysis of eosinophil mRNA. Results: Both peripheral blood and bronchoalveolar lavage fluid eosinophils expressed NFATp and NFATc protein. Unlike activated T cells, which express multiple NFATc isoforms, eosinophils preferentially express the approximately 85-kd isoform. In addition, eosinophils were found to constitutively express NFATc mRNA. A brief incubation with the TH2 cytokines IL-4 and IL-5 was sufficient to induce the nuclear translocation of NFATc. Eosinophil nuclear extracts contain multiple factors that can specifically recognize the IL-4 promoter P1 NFAT site in DNA-binding assays, including NFATp. Conclusion: NFATp and NFATc can regulate the expression of cytokines and other genes in eosinophils but appear to be regulated by a novel signal transduction mechanism in these cells.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 143-152 |
| Number of pages | 10 |
| Journal | The Journal of Allergy and Clinical Immunology |
| Volume | 107 |
| Issue number | 1 |
| DOIs | |
| State | Published - 2001 |
| Externally published | Yes |
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Keywords
- Cytokines
- Eosinophils
- HMG1/Y
- Nuclear factor of activated T cells
- Transcriptional regulation
ASJC Scopus subject areas
- Immunology and Allergy
- Immunology
Cite this
Human eosinophils constitutively express nuclear factor of activated T cells p and c. / Seminario, Maria Cristina; Guo, Jia; Bochner, Bruce S.; Beck, Lisa A.; Georas, Steve N.
In: The Journal of Allergy and Clinical Immunology, Vol. 107, No. 1, 2001, p. 143-152.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Human eosinophils constitutively express nuclear factor of activated T cells p and c
AU - Seminario, Maria Cristina
AU - Guo, Jia
AU - Bochner, Bruce S.
AU - Beck, Lisa A.
AU - Georas, Steve N.
PY - 2001
Y1 - 2001
N2 - Background: Eosinophils are now known to produce a variety of proinflammatory cytokines, although the molecular factors that regulate their production are poorly understood. The expression of almost all of the cytokines produced by eosinophils, including the proallergic cytokine IL-4, is now known to be regulated at the level of transcription by members of the nuclear factor of activated T cells (NFAT) family of transcription factors. Objective: We sought to characterize the expression of different NFAT proteins in resting and activated eosinophils. Methods: Nuclear and whole cell extracts were obtained from both peripheral blood eosinophils and those obtained from bronchoalveolar lavage fluid of asthmatic subjects after endobronchial allergen challenge. NFAT expression was determined by using immunoprecipitation and Western blot analysis, DNA-binding assays, and RT-PCR analysis of eosinophil mRNA. Results: Both peripheral blood and bronchoalveolar lavage fluid eosinophils expressed NFATp and NFATc protein. Unlike activated T cells, which express multiple NFATc isoforms, eosinophils preferentially express the approximately 85-kd isoform. In addition, eosinophils were found to constitutively express NFATc mRNA. A brief incubation with the TH2 cytokines IL-4 and IL-5 was sufficient to induce the nuclear translocation of NFATc. Eosinophil nuclear extracts contain multiple factors that can specifically recognize the IL-4 promoter P1 NFAT site in DNA-binding assays, including NFATp. Conclusion: NFATp and NFATc can regulate the expression of cytokines and other genes in eosinophils but appear to be regulated by a novel signal transduction mechanism in these cells.
AB - Background: Eosinophils are now known to produce a variety of proinflammatory cytokines, although the molecular factors that regulate their production are poorly understood. The expression of almost all of the cytokines produced by eosinophils, including the proallergic cytokine IL-4, is now known to be regulated at the level of transcription by members of the nuclear factor of activated T cells (NFAT) family of transcription factors. Objective: We sought to characterize the expression of different NFAT proteins in resting and activated eosinophils. Methods: Nuclear and whole cell extracts were obtained from both peripheral blood eosinophils and those obtained from bronchoalveolar lavage fluid of asthmatic subjects after endobronchial allergen challenge. NFAT expression was determined by using immunoprecipitation and Western blot analysis, DNA-binding assays, and RT-PCR analysis of eosinophil mRNA. Results: Both peripheral blood and bronchoalveolar lavage fluid eosinophils expressed NFATp and NFATc protein. Unlike activated T cells, which express multiple NFATc isoforms, eosinophils preferentially express the approximately 85-kd isoform. In addition, eosinophils were found to constitutively express NFATc mRNA. A brief incubation with the TH2 cytokines IL-4 and IL-5 was sufficient to induce the nuclear translocation of NFATc. Eosinophil nuclear extracts contain multiple factors that can specifically recognize the IL-4 promoter P1 NFAT site in DNA-binding assays, including NFATp. Conclusion: NFATp and NFATc can regulate the expression of cytokines and other genes in eosinophils but appear to be regulated by a novel signal transduction mechanism in these cells.
KW - Cytokines
KW - Eosinophils
KW - HMG1/Y
KW - Nuclear factor of activated T cells
KW - Transcriptional regulation
UR - http://www.scopus.com/inward/record.url?scp=0035144977&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0035144977&partnerID=8YFLogxK
U2 - 10.1067/mai.2001.111931
DO - 10.1067/mai.2001.111931
M3 - Article
C2 - 11150004
AN - SCOPUS:0035144977
VL - 107
SP - 143
EP - 152
JO - Journal of Allergy and Clinical Immunology
JF - Journal of Allergy and Clinical Immunology
SN - 0091-6749
IS - 1
ER -