Gene expression analysis of Tec family kinases in B- and T-lymphocytes
Author: Blomberg, K Emelie M
Date: 2009-06-12
Location: C1.87, Karolinska Universitetssjukhuset, Huddinge
Time: 09.00
Department: Institutionen för laboratoriemedicin / Department of Laboratory Medicine
View/ Open:
Thesis (734.4Kb)
Abstract
The microarray technology is a powerful tool used in many different
research areas. The technique has been around for more than a decade and
has revolutionized the molecular biology field. In this thesis the
Affymetrix GeneChip® arrays were used to study the transcriptomes of Tec
family kinase mutant mouse models. Bruton s tyrosine kinase (Btk), IL-2
inducible T-cell kinase (Itk) and Tyrosine kinase expressed in
hepatocellular carcinoma (Tec) are protein tyrosine kinases belonging to
this Tec kinase family. Mutations in Btk cause a primary immunodeficiency
disease called Xlinked agammaglobulinemia (XLA) in humans and X-linked
immunodeficiencydisease (Xid) in mice. Gene expression profiling was
performed on whole splenic Bcells as well as Transitional 1 (T1) B-cells
from Xid, Btk knockout (Btk KO) and control mice. This was done in order
to study differences and similarities between Btkdefective mice (Xid and
Btk KO together) compared to control. Small differences were
distinguished between the Btk-defective mouse strains in the whole
splenic B-cell population; only seven genes differed (>2-fold) between
the two. A number of potentially interesting genes were found to be
differentially expressed in the Btkdefective groups compared to the
controls. We also show that the Btk defect is already manifested at the
T1 B-cell stage.
Itk is important for the T-cell development where it has a role in regulating the conventional versus innate T-cells. Itk-deficient T-cells are of an innate, memory-like type. Tec is poorly expressed in resting T-cells, but is expressed 2-3 days after stimulation. Tec-deficient mice show no phenotypic alterations and the mice develop normally. In Papers III and IV we looked at the transcriptomes of Itk-, Tec- and Itk/Tec-deficient mice by studying unstimulated as well as stimulated CD3+ T-cells. The Itk-deficiency was also studied in CD4+ and CD8+ T-cell subpopulations. The gene expression patterns of Tec-deficient mice compared to control mice showed small differences, further supporting earlier findings. More differences were seen in Itkdeficient as well as Itk/Tec-deficient mice compared to controls. We also investigated if the Itk-deficiency could mimic calcineurin inhibition by treating Wt T-cells with cyclosporin A (CsA). CsA was shown to have a stronger effect on transcriptional regulation than Itk-deficiency, suggesting that only a fraction of TCR-mediated calcineurin/NFAT-activation is dependent on Itk. Greater differences were seen in CD4+ and CD8+ T-cells in comparison to total CD3+ T-cells. In Paper IV we found Zbtb26, an interesting transcription factor, being up-regulated in Itk-deficient T-cells, while normalized in Itk/Tec-deficient cells compared to Wt. By the use of a combination of the microarray technology and quantitative RT-PCR analysis, we have evaluated the transcriptomes of Btk-, Itk-, Tec- and Itk/Tec-deficient mice.
Itk is important for the T-cell development where it has a role in regulating the conventional versus innate T-cells. Itk-deficient T-cells are of an innate, memory-like type. Tec is poorly expressed in resting T-cells, but is expressed 2-3 days after stimulation. Tec-deficient mice show no phenotypic alterations and the mice develop normally. In Papers III and IV we looked at the transcriptomes of Itk-, Tec- and Itk/Tec-deficient mice by studying unstimulated as well as stimulated CD3+ T-cells. The Itk-deficiency was also studied in CD4+ and CD8+ T-cell subpopulations. The gene expression patterns of Tec-deficient mice compared to control mice showed small differences, further supporting earlier findings. More differences were seen in Itkdeficient as well as Itk/Tec-deficient mice compared to controls. We also investigated if the Itk-deficiency could mimic calcineurin inhibition by treating Wt T-cells with cyclosporin A (CsA). CsA was shown to have a stronger effect on transcriptional regulation than Itk-deficiency, suggesting that only a fraction of TCR-mediated calcineurin/NFAT-activation is dependent on Itk. Greater differences were seen in CD4+ and CD8+ T-cells in comparison to total CD3+ T-cells. In Paper IV we found Zbtb26, an interesting transcription factor, being up-regulated in Itk-deficient T-cells, while normalized in Itk/Tec-deficient cells compared to Wt. By the use of a combination of the microarray technology and quantitative RT-PCR analysis, we have evaluated the transcriptomes of Btk-, Itk-, Tec- and Itk/Tec-deficient mice.
List of papers:
I. Lindvall JM, Blomberg KE, Berglöf A, Yang Q, Smith CI, Islam TC (2004). "Gene expression profile of B cells from Xid mice and Btk knockout mice." Eur J Immunol 34(7): 1981-91
Pubmed
II. Lindvall JM, Blomberg KE, Berglöf A, Smith CI (2006). "Distinct gene expression signature in Btk-defective T1 B-cells." Biochem Biophys Res Commun 346(2): 461-9. Epub 2006 Jun 2
Pubmed
III. Blomberg KE, Boucheron N, Lindvall JM, Yu L, Raberger J, Berglof A, Ellmeier W, Smith CI (2009). "Transcriptional signatures of Itk-deficient CD3+, CD4+ and CD8+ T-cells." BMC Genomics 10(1): 233. [Epub ahead of print]
Pubmed
IV. Blomberg KEM, Boucheron N, Lindvall JM, Vargas L, Raberger J, Berglöf A, Ellmeier W, Smith CIE (2009). "Tec and Itk regulate overlapping signaling pathways in T lymphocytes." (Manuscript)
I. Lindvall JM, Blomberg KE, Berglöf A, Yang Q, Smith CI, Islam TC (2004). "Gene expression profile of B cells from Xid mice and Btk knockout mice." Eur J Immunol 34(7): 1981-91
Pubmed
II. Lindvall JM, Blomberg KE, Berglöf A, Smith CI (2006). "Distinct gene expression signature in Btk-defective T1 B-cells." Biochem Biophys Res Commun 346(2): 461-9. Epub 2006 Jun 2
Pubmed
III. Blomberg KE, Boucheron N, Lindvall JM, Yu L, Raberger J, Berglof A, Ellmeier W, Smith CI (2009). "Transcriptional signatures of Itk-deficient CD3+, CD4+ and CD8+ T-cells." BMC Genomics 10(1): 233. [Epub ahead of print]
Pubmed
IV. Blomberg KEM, Boucheron N, Lindvall JM, Vargas L, Raberger J, Berglöf A, Ellmeier W, Smith CIE (2009). "Tec and Itk regulate overlapping signaling pathways in T lymphocytes." (Manuscript)
Issue date: 2009-05-22
Rights:
Publication year: 2009
ISBN: 978-91-7409-452-7
Statistics
Total Visits
Views | |
---|---|
Gene ...(legacy) | 591 |
Gene ... | 112 |
Total Visits Per Month
October 2023 | November 2023 | December 2023 | January 2024 | February 2024 | March 2024 | April 2024 | |
---|---|---|---|---|---|---|---|
Gene ... | 4 | 0 | 0 | 0 | 6 | 0 | 0 |
File Visits
Views | |
---|---|
thesis.pdf(legacy) | 558 |
thesis.pdf | 232 |
thesis.pdf.txt(legacy) | 2 |
Top country views
Views | |
---|---|
United States | 347 |
China | 60 |
Germany | 47 |
Sweden | 43 |
South Korea | 28 |
Finland | 15 |
Denmark | 13 |
Japan | 10 |
Russia | 10 |
United Kingdom | 9 |
Top cities views
Views | |
---|---|
Romeo | 28 |
Sunnyvale | 28 |
Beijing | 23 |
Kiez | 17 |
Seoul | 14 |
Ballerup | 13 |
Dublin | 8 |
Helsinki | 7 |
London | 6 |
University Park | 6 |