Assuntos
Receptores de Antígenos de Linfócitos T gama-delta/fisiologia , Subpopulações de Linfócitos T/imunologia , Animais , Butirofilinas/fisiologia , Congressos como Assunto , Humanos , Imunoterapia , Infecções/imunologia , Inflamação/imunologia , Ativação Linfocitária , Neoplasias/imunologia , Receptores de Retorno de Linfócitos/fisiologia , Subpopulações de Linfócitos T/fisiologiaRESUMO
Human Vγ9Vδ2 T cells respond to microbial infections as well as certain types of tumors. The key initiators of Vγ9Vδ2 activation are small, pyrophosphate-containing molecules called phosphoantigens (pAgs) that are present in infected cells or accumulate intracellularly in certain tumor cells. Recent studies demonstrate that initiation of the Vγ9Vδ2 T cell response begins with sensing of pAg via the intracellular domain of the butyrophilin 3A1 (BTN3A1) molecule. However, it is unknown how downstream events can ultimately lead to T cell activation. Here, using NMR spectrometry and molecular dynamics (MD) simulations, we characterize a global conformational change in the B30.2 intracellular domain of BTN3A1 induced by pAg binding. We also reveal by crystallography two distinct dimer interfaces in the BTN3A1 full-length intracellular domain, which are stable in MD simulations. These interfaces lie in close proximity to the pAg-binding pocket and contain clusters of residues that experience major changes of chemical environment upon pAg binding. This suggests that pAg binding disrupts a preexisting conformation of the BTN3A1 intracellular domain. Using a combination of biochemical, structural, and cellular approaches we demonstrate that the extracellular domains of BTN3A1 adopt a V-shaped conformation at rest, and that locking them in this resting conformation without perturbing their membrane reorganization properties diminishes pAg-induced T cell activation. Based on these results, we propose a model in which a conformational change in BTN3A1 is a key event of pAg sensing that ultimately leads to T cell activation.
Assuntos
Antígenos CD/fisiologia , Butirofilinas/fisiologia , Linfócitos Intraepiteliais/efeitos dos fármacos , Antígenos/imunologia , Antígenos CD/química , Antígenos CD/metabolismo , Butirofilinas/química , Cristalografia por Raios X , Células HEK293 , Humanos , Linfócitos Intraepiteliais/fisiologia , Ativação Linfocitária/imunologia , Ativação Linfocitária/fisiologia , Espectroscopia de Ressonância Magnética/métodos , Fosforilação , Ligação Proteica , Conformação Proteica , Domínios Proteicos , Receptores de Antígenos de Linfócitos T gama-delta/imunologia , Relação Estrutura-Atividade , Linfócitos T/imunologiaRESUMO
Mammalian butyrophilins have various important functions, one for lipid binding but others as ligands for co-inhibition of αß T cells or for stimulation of γδ T cells in the immune system. The chicken BG homologues are dimers, with extracellular immunoglobulin variable (V) domains joined by cysteines in the loop equivalent to complementarity-determining region 1 (CDR1). BG genes are found in three genomic locations: BG0 on chromosome 2, BG1 in the classical MHC (the BF-BL region) and many BG genes in the BG region just outside the MHC. Here, we show that BG0 is virtually monomorphic, suggesting housekeeping function(s) consonant with the ubiquitous tissue distribution. BG1 has allelic polymorphism but minimal sequence diversity, with the few polymorphic residues at the interface of the two V domains, suggesting that BG1 is recognized by receptors in a conserved fashion. Any phenotypic variation should be due to the intracellular region, with differential exon usage between alleles. BG genes in the BG region can generate diversity by exchange of sequence cassettes located in loops equivalent to CDR1 and CDR2, consonant with recognition of many ligands or antigens for immune defence. Unlike the mammalian butyrophilins, there are at least three modes by which BG genes evolve.