Knockout B lymphoma cell lines as biochemical tools to explore multiple signalling pathways
- Author(s)
- Veale, MF; Dietrich, WM; Corcoran, LM;
- Details
- Publication Year 2003-08,Volume 81,Issue #4,Page 297-304
- Journal Title
- IMMUNOLOGY AND CELL BIOLOGY
- Publication Type
- Journal Article
- Abstract
- Studies on B lymphocyte signalling pathways using B lymphocytes from genetically modified mice have the disadvantages of primary cell polyclonality and finite life span. B lymphoma cell lines have been generated from mice with targeted mutations in the oct-2, OBF-1, vav-1 and btk genes, as a model system that lacks these limitations and possesses additional potential for experimental manipulation. To assess their utility, activation of the B cell receptor using anti-mu, the Toll-like receptor-4 using lipopolysaccharide and the interleukin-4 receptor were assessed in these cell lines. Differential tyrosine phosphorylation of intracellular proteins was measured in the wild-type controls compared to the corresponding mutant cell lines after B cell receptor stimulation. Intracellular calcium (Ca-i(2+)) was mobilized in the control cell lines but not in the OBF-1 and Vav1-deficient cells, while Xid B cell lines (btk mutant) showed a reduced Ca2+ mobilization. Extracellular signal-regulated kinase 1/2 phosphorylation in response to anti-mu or lipopolysaccharide stimulation was significantly reduced in Vav1-deficient cells. Interleukin-4 stimulation of wild-type cells resulted in a 2-3-fold increase in Stat-6 phosphorylation. These results indicate that the cell lines mimic the biochemical responses of the corresponding primary B cells. They therefore represent a useful model system to investigate the regulation and roles of these and other gene products in B cell signal transduction and activation.
- Publisher
- BLACKWELL PUBLISHING ASIA
- Keywords
- RECEPTOR-MEDIATED ACTIVATION; PROTEIN-TYROSINE KINASES; JUN NH2-TERMINAL KINASE; ANTIGEN-RECEPTOR; REGULATED KINASE; T-CELL; PHOSPHATIDYLINOSITOL 3-KINASE; TRANSCRIPTION FACTOR; CROSS-LINKING; FC-RECEPTOR
- Publisher's Version
- https://doi.org/10.1046/j.0818-9641.2003.01172.x
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- Refer to copyright notice on published article.
Creation Date: 2003-08-01 12:00:00