Producción CyT
Congreso
Autoría
Pariani, Alejandro P.
;
Almada, Evangelina
;
Hidalgo, Florencia
;
MARIN, MARINA LEANDRA
;
Favre, Cristian
;
Cecilia Larocca, Maria
Fecha
2019
Editorial y Lugar de Edición
SAFIS
Resumen
Información suministrada por el agente en
SIGEVA
The immune synapse (IS) refers to a specialized cell-cell junction established between an immune cell and its target cell (TC). When a natural killer cell (NK) is activated by a susceptible TC, the actin cytoskeleton reorganizes at the IS, facilitating integrin and receptor clustering. Subsequently, the centrosome polarizes to the IS, enabling the directional secretion of cytotoxic components, which triggers the lysis of the TC. AKAP350 is an A-kinase anchoring protein essential in the regulati...
The immune synapse (IS) refers to a specialized cell-cell junction established between an immune cell and its target cell (TC). When a natural killer cell (NK) is activated by a susceptible TC, the actin cytoskeleton reorganizes at the IS, facilitating integrin and receptor clustering. Subsequently, the centrosome polarizes to the IS, enabling the directional secretion of cytotoxic components, which triggers the lysis of the TC. AKAP350 is an A-kinase anchoring protein essential in the regulation ofmicrotubule dynamics. AKAP350 interacts with the microtubule plus end binding protein 1 (EB1), which participates in the crosstalk between microtubule and actin cytoskeleton. In non-lytic T cell IS, AKAP350 participates in LFA-1 clustering, whereas EB1 is involved in T cell receptor activation. Our previous results indicate that AKAP350 participates in NK cytotoxic response, by regulating actin accumulation and LFA-1 clustering at the IS. The aim of our work was to demonstrate if AKAP350 participation occurs exclusively downstream LFA-1 activation and to evaluate EB1 as a possible mediator. YTS NK cells with decreased expression of AKAP350 (AKAP350KD) were activated for 30 min using specific ligands for LFA-1 or CD28. Actin accumulation and LFA-1clustering were analyzed by confocal microscopy. Actin accumulation at the IS was impaired in AKAP350KD cells activated via LFA-1 (-37%*) or CD28 (-73%*). Concomitantly, AKAP350KD cells showed decreased LFA-1 localization at the IS in cells activated via either LFA-1 (-40%*) or CD28 (-60 %*). In order to analyze AKAP350 role in EB1 recruitment to microtubules at the IS, YTSAKAP350KD cells were incubated with KT86 cells (TC) for 30 min. EB1 localization was analyzed by confocal microscopy. YTS AKAP350KD showed decreased EB1 localization at the IS edge (-45%*) and at IS microtubules (-55%*). The latter results were confirmed by western blot analysis of microtubule fractions of YTS cells activated via LFA-1. Overall, our results suggest that AKAP350participates in actin accumulation and receptor clustering downstream both LFA-1 and CD28 activations and position EB1 as a putative mediator of this effect. *p<0.05.
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Palabras Clave
AKAP350Golgi apparatusImmune synapseAKAP450