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Flow cytometric measurement of cytosolic calcium in lymphocytes

par Baus, Erika ;Urbain, Jacques ;Leo, Oberdan ;Andris, Fabienne
Editeur scientifique van Duijn, Bert;Wiltink, Anneke
Référence Signal Transduction- Single Cell Techniques, Springer-Verlag, Berlin Heidelberg, page (358-372)
Publication Publié, 1998

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Résumé :

Calcium plays a central role in the transduction of external stimuli in many cell types. In B and T lymphocytes, stimulation of surface immunoglobulins or T cell receptors, respectively, triggers the activation of members of the src family of tyrosine kinases. These enzymes phosphorylate several intracellular substrates including phosphoinositidespecific phosphodiesterase (PLCγ), GTPase-activating protein for p2lras (GAP) and phosphatidylinositol 3-kinase (PI3-kinase). PLCγ hydrolyzes phosphatidylinositol 4,5-bisphosphate (PIP2) into diacylglycerol and inositol trisphosphate. These second messengers are responsible for the activation of protein kinase C (PKC) and an increase in the concentration of intracellular ionized calcium ([Ca2+]i). Together, these agents are capable of initiating a cascade of biochemical events that results in new gene expression leading to functional responses such as cytokine production, up-regulation of cytokine receptor expression and cellular proliferation (for a review of this subject see Weiss and Littman 1994). The role of calcium in the response of lymphocytes to extracellular stimuli is clearly demonstrated by the observation that cells cultured in medium containing low levels of extracellular calcium fail to proliferate or produce cytokines upon receptor cross-linking (Mills et al. 1985; Dennis et al. 1987).