Producción CyT
VALIDATE NETWORK - Comparative transcriptomic profiles of bovine differentiated macrophages and autologous lymphocytes co-cultures infected with two M. bovis isolates of opposite virulence phenotypes reveals a differential up regulation of type I IFN pathway genes.
Congreso
Autoría:
Blanco, Federico Carlos ; Bigi, María M. ; GARCIA, ELIZABETH ANDREA ; Vazquez, Cristina Lourdes ; Bigi, FabianaFecha:
2023Editorial y Lugar de Edición:
VALIDATEResumen *
The M. bovis Mb04-303 isolate was previously characterized by our group using in vitro and in vivo models. This wild board isolate of M. bovis is able to produce extended granulomatous lesions andsevere pathology in cattle as well as in the mouse and guinea pig model. In a recent publication, we have described the innate immune response elicited by Mb04-303 using an in vitro co-culturemodel of bovine differentiated macrophages and autologous lymphocytes. In comparison with other two M. bovis isolates, Mb534 and NCTC10772, the innate immune response elicited by Mb04-303 was the only capable of inhibit intracellular mycobacterial replication and was characterized by increased percentages of NKp46+IFNɣ+ cells. In order to decipher the immune mediators involved in this efficient innate immune response we performed transcriptomic analysis of co-cultures infected with Mb534 and Mb04-303, the two strains with opposite virulence phenotypes. Results of the RNA sequencing showed 237 differentially expressed genes (DEGs). Remarkably, the most upregulated genes in co-cultures infected with Mb04-303 are related to the type I IFN activation pathway, as well as several IFN induced genes like RSAD2, ISG15, Mx1, IFIT2 and ISG20. Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) analysis of the RNAseq data showed DEGs networks related to Innate Immune Response, Cellular Immune Response, Antiviral Immune Response and Type I IFN pathway activation. This particular gene expression signature is characterized by recognition of mycobacteria by intracellular pattern recognition receptors (PRRs). Since induction of Type I IFN pathway has been associated to the phagosome escape of Mycobacterium abscessus (doi: 10.3389/fimmu.2019.00125) we evaluated the M. bovis cytoplasmatic localization. For this purpose, we performed infections of primary macrophages with Mb04-303 or Mb534 and analyzed intracellular trafficking using confocal microscopy and indirect fluorescence staining of the cytoplasmatic marker Galectin8. We observed significant more colocalization of Mb04-303 with Galectin8 than with Mb534. This preliminary result indicate that Mb04-303 evades the phagolysosomal pathway, escapes into the cellular cytoplasm and potentially increases the interaction with intracellular PRRs more efficiently than Mb534 Información suministrada por el agente en SIGEVAPalabras Clave
TRANSCRIPTOMICMYCOBACTERIUM BOVISIFN