Back to Search
Start Over
Analysis of TGFβ1 and IL-10 transcriptional regulation in CTCL cells by chromatin immunoprecipitation.
- Source :
-
Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2014; Vol. 1172, pp. 329-41. - Publication Year :
- 2014
-
Abstract
- The immunosuppressive cytokines transforming growth factor β1 (TGFβ1) and interleukin-10 (IL-10) regulate a variety of biological processes including differentiation, proliferation, tissue repair, tumorigenesis, inflammation, and host defense. Aberrant expression of TGFβ1 and IL-10 has been associated with many types of autoimmune and inflammatory disorders, as well as with many types of cancer and leukemia. Patients with cutaneous T cell lymphoma (CTCL) have high levels of malignant CD4+ T cells expressing IL-10 and TGFβ1 that suppress the immune system and diminish the antitumor responses. The transcriptional regulation of TGFβ1 and IL-10 expression is orchestrated by several transcription factors, including NFκB. However, while the transcriptional regulation of pro-inflammatory and anti-apoptotic genes by NFκB has been studied extensively, much less is known about the NFκB regulation of immunosuppressive genes. In this chapter, we describe a protocol that uses chromatin immunoprecipitation (ChIP) to analyze the transcriptional regulation of TGFβ1 and IL-10 by measuring recruitment of NFκB p65, p50, c-Rel, Rel-B, and p52 subunits to TGFβ1 and IL-10 promoters in human CTCL Hut-78 cells.
- Subjects :
- CD4-Positive T-Lymphocytes pathology
Chromatin Immunoprecipitation
Humans
Interleukin-10 metabolism
Lymphoma, T-Cell, Cutaneous metabolism
Lymphoma, T-Cell, Cutaneous pathology
NF-kappa B p50 Subunit genetics
NF-kappa B p50 Subunit metabolism
NF-kappa B p52 Subunit genetics
NF-kappa B p52 Subunit metabolism
Promoter Regions, Genetic
Proto-Oncogene Proteins c-rel genetics
Proto-Oncogene Proteins c-rel metabolism
Real-Time Polymerase Chain Reaction
Signal Transduction
Transcription Factor RelA genetics
Transcription Factor RelA metabolism
Transcription Factor RelB genetics
Transcription Factor RelB metabolism
Transcription, Genetic
Transforming Growth Factor beta1 metabolism
Tumor Cells, Cultured
CD4-Positive T-Lymphocytes metabolism
Interleukin-10 genetics
Lymphoma, T-Cell, Cutaneous genetics
Transforming Growth Factor beta1 genetics
Subjects
Details
- Language :
- English
- ISSN :
- 1940-6029
- Volume :
- 1172
- Database :
- MEDLINE
- Journal :
- Methods in molecular biology (Clifton, N.J.)
- Publication Type :
- Academic Journal
- Accession number :
- 24908319
- Full Text :
- https://doi.org/10.1007/978-1-4939-0928-5_30