RESUMEN
Innate lymphoid cell (ILC) development proposes that ILC precursors (ILCPs) segregate along natural killer (NK) cell versus helper cell (ILC1, ILC2, ILC3) pathways, the latter depending on expression of Id2, Zbtb16, and Gata3. We have developed an Id2-reporter strain expressing red fluorescent protein (RFP) in the context of normal Id2 expression to re-examine ILCP phenotype and function. We show that bone-marrow ILCPs were heterogeneous and harbored extensive NK-cell potential in vivo and in vitro. By multiplexing Id2RFP with Zbtb16CreGFP and Bcl11btdTomato strains, we made a single-cell dissection of the ILCP compartment. In contrast with the current model, we have demonstrated that Id2+Zbtb16+ ILCPs included multi-potent ILCPs that retained NK-cell potential. Late-stage ILC2P and ILC3P compartments could be defined by differential Zbtb16 and Bcl11b expression. We suggest a revised model for ILC differentiation that redefines the cell-fate potential of helper-ILC-restricted Zbtb16+ ILCPs.
Asunto(s)
Regulación de la Expresión Génica/inmunología , Células Madre Hematopoyéticas/citología , Inmunidad Innata , Proteína 2 Inhibidora de la Diferenciación/genética , Linfopoyesis/genética , Traslado Adoptivo , Animales , Linaje de la Célula , Factor de Transcripción GATA3/biosíntesis , Factor de Transcripción GATA3/genética , Factor de Transcripción GATA3/fisiología , Genes Reporteros , Células Madre Hematopoyéticas/metabolismo , Proteína 2 Inhibidora de la Diferenciación/biosíntesis , Células Asesinas Naturales/citología , Proteínas Luminiscentes/análisis , Proteínas Luminiscentes/genética , Ratones , Ratones Endogámicos C57BL , Modelos Inmunológicos , Proteína de la Leucemia Promielocítica con Dedos de Zinc/biosíntesis , Proteína de la Leucemia Promielocítica con Dedos de Zinc/genética , Proteína de la Leucemia Promielocítica con Dedos de Zinc/fisiología , Análisis de la Célula Individual , Linfocitos T Colaboradores-Inductores/citología , Transcripción Genética , Proteína Fluorescente RojaRESUMEN
Because human corneal endothelial cells (HCECs) do not proliferate once the endothelial monolayer has formed, corneal wound healing is believed to be mediated by cell enlargement or migration, rather than by proliferation. However, the cellular mechanisms involved in wound healing by HCECs have not been fully determined. In this review, we focus on the effects of promyelocytic leukemia zinc finger (PLZF), a DNA-binding transcription factor, and transforming growth factor (TGF)-ß2 on the proliferation and migration of cultured HCECs. Involvement of the mitogen-activated protein kinase (MAPK) signaling pathway in the migration of HCECs was also investigated. Expression of PLZF mRNA decreased as cell-cell contact was disrupted and returned to the original level as cell-cell contact was re-formed. Assessment with a real-time cell electronic sensing system revealed that proliferation of cultured HCECs was inhibited after infection with Ad-PLZF and exposure to TGF-ß2. Migration of cultured HCECs was increased by TGF-ß2 through p38 MAPK activation. We conclude that PLZF expression in cultured HCECs is closely related to the formation of cell-cell contact and that TGF-ß2 suppresses proliferation of cultured HCECs, while promoting their migration through p38 MAPK activation.