RESUMO
Proliferation of T cells via activation of the T-cell receptor (TCR) requires concurrent engagement of accessory costimulatory molecules to achieve full activation. The best-studied costimulatory molecule, CD28, achieves these effects, in part, by augmenting signals from the TCR to the mitogen-activated protein (MAP) kinase cascade. We show here that TCR-mediated stimulation of MAP kinase extracellular-signal-regulated kinases (ERKs) is limited by activation of the Ras antagonist Rap1. CD28 increases ERK signaling by blocking Rap1 action. CD28 inhibits Rap1 activation because it selectively stimulates an extrinsic Rap1 GTPase activity. The ability of CD28 to stimulate Rap1 GTPase activity was dependent on the tyrosine kinase Lck. Our results suggest that CD28-mediated Rap1 GTPase-activating protein activation can help explain the augmentation of ERKs during CD28 costimulation.
Assuntos
Antígenos CD28/metabolismo , Proteína Tirosina Quinase p56(lck) Linfócito-Específica/metabolismo , Quinases de Proteína Quinase Ativadas por Mitógeno/metabolismo , Linfócitos T/metabolismo , Proteínas rap1 de Ligação ao GTP/metabolismo , Animais , Complexo CD3/metabolismo , Ativação Enzimática/efeitos dos fármacos , Inibidores Enzimáticos/farmacologia , Flavonoides/farmacologia , Humanos , Células Jurkat/efeitos dos fármacos , Células Jurkat/metabolismo , Proteína Tirosina Quinase p56(lck) Linfócito-Específica/genética , Sistema de Sinalização das MAP Quinases , Camundongos , Camundongos Endogâmicos C57BL , Quinases de Proteína Quinase Ativadas por Mitógeno/antagonistas & inibidores , Receptores de Antígenos de Linfócitos T/efeitos dos fármacos , Receptores de Antígenos de Linfócitos T/metabolismo , Proteínas ras/genética , Proteínas ras/metabolismo , Domínios de Homologia de srcRESUMO
Over the last decade the concept of T cell co-stimulation has emerged to take a central role in the process of T cell activation. However, the exact definition of co-stimulation is still unclear. In this review, we re-examine the concept of co-stimulation. We suggest that while co-stimulation is important, there is little evidence to link co-stimulation with T cell anergy. We then suggest a framework for studying co-stimulation. Focusing on recent advances in our understanding of CD28, we discuss four areas of T cell activation where co-stimulation may play a role.
Assuntos
Antígenos CD28/imunologia , Ativação Linfocitária/imunologia , Transdução de Sinais/imunologia , Linfócitos T/imunologia , Animais , Sobrevivência Celular/imunologia , Humanos , Interleucina-2/genética , Interleucina-2/metabolismo , Camundongos , NF-kappa B/imunologia , Regiões Promotoras GenéticasRESUMO
The Src family tyrosine kinases Lck and Fyn are critical for signaling via the T cell receptor. However, the exact mechanism of their activation is unknown. Recent crystal structures of Src kinases suggest that an important mechanism of kinase activation is via engagement of the Src homology (SH)3 domain by proline-containing sequences. To test this hypothesis, we identified several T cell membrane proteins that contain potential SH3 ligands. Here we demonstrate that Lck and Fyn can be activated by proline motifs in the CD28 and CD2 proteins, respectively. Supporting a role for Lck in CD28 signaling, we demonstrate that CD28 signaling in both transformed and primary T cells requires Lck as well as proline residues in CD28. These data suggest that Lck plays an essential role in CD28 costimulation.
Assuntos
Antígenos CD28/fisiologia , Ativação Linfocitária , Proteína Tirosina Quinase p56(lck) Linfócito-Específica/imunologia , Prolina/fisiologia , Linfócitos T/imunologia , Domínios de Homologia de src/imunologia , Alanina/imunologia , Sequência de Aminoácidos , Substituição de Aminoácidos/imunologia , Animais , Antígenos CD28/genética , Antígenos CD28/metabolismo , Ativação Enzimática/imunologia , Regulação da Expressão Gênica/imunologia , Genes fos/imunologia , Humanos , Células Jurkat , Ativação Linfocitária/efeitos dos fármacos , Ativação Linfocitária/genética , Proteína Tirosina Quinase p56(lck) Linfócito-Específica/deficiência , Proteína Tirosina Quinase p56(lck) Linfócito-Específica/genética , Camundongos , Camundongos Endogâmicos C57BL , Dados de Sequência Molecular , Peptídeos/antagonistas & inibidores , Peptídeos/síntese química , Peptídeos/imunologia , Prolina/deficiência , Prolina/genética , Ligação Proteica/imunologia , Proteínas Tirosina Quinases/metabolismo , Proteínas Proto-Oncogênicas/metabolismo , Proteínas Proto-Oncogênicas c-fyn , Retroviridae/genética , Retroviridae/imunologia , Linfócitos T/metabolismo , Linfócitos T/virologia , Acetato de Tetradecanoilforbol/farmacologiaRESUMO
Recognition of antigen by T cells requires the formation of a specialized junction between the T cell and the antigen-presenting cell. This junction is generated by the recruitment and the exclusion of specific proteins from the contact area. The mechanisms that regulate these events are unknown. Here we demonstrate that ligand engagement of the adhesion molecule, CD2, initiates a process of protein segregation, CD2 clustering, and cytoskeletal polarization. Although protein segregation was not dependent on the cytoplasmic domain of CD2, CD2 clustering and cytoskeletal polarization required an interaction of the CD2 cytoplasmic domain with a novel SH3-containing protein. This novel protein, called CD2AP, is likely to facilitate receptor patterning in the contact area by linking specific adhesion receptors to the cytoskeleton.