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Requirement of Transmembrane Domain for CD154 Association to Lipid Rafts and Subsequent Biological Events

Abstract : Interaction of CD40 with CD154 leads to recruitment of both molecules into lipid rafts, resulting in bi-directional cell activation. The precise mechanism by which CD154 is translocated into lipid rafts and its impact on CD154 signaling remain largely unknown. Our aim is to identify the domain of CD154 facilitating its association to lipid rafts and the impact of such association on signaling events and cytokine production. Thus, we generated Jurkat cell lines expressing truncated CD154 lacking the cytoplasmic domain or chimeric CD154 in which the transmembrane domain was replaced by that of transferrin receptor I, known to be excluded from lipid rafts. Our results show that cell stimulation with soluble CD40 leads to the association of CD154 wild-type and CD154-truncated, but not CD154-chimera, with lipid rafts. This is correlated with failure of CD154-chimera to activate Akt and p38 MAP kinases, known effectors of CD154 signaling. We also found that CD154-chimera lost the ability to promote IL-2 production upon T cell stimulation with anti-CD3/CD28 and soluble CD40. These results demonstrate the implication of the transmembrane domain of CD154 in lipid raft association, and that this association is necessary for CD154-mediated Akt and p38 activation with consequent enhancement of IL-2 production.
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Nadir Benslimane, Ghada Hassan, Daniel Yacoub, Walid Mourad. Requirement of Transmembrane Domain for CD154 Association to Lipid Rafts and Subsequent Biological Events. PLoS ONE, Public Library of Science, 2012, 7 (8), pp. e43070. ⟨10.1371/journal.pone.0043070.g005⟩. ⟨hal-01537234⟩



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