Citation

  • Authors: Jia Z. et al.
  • Year: 2023
  • Journal: Dev Comp Immunol 141 104632
  • Applications: in vitro / DNA / jetOPTIMUS
  • Cell type: HEK-293
    Description: Human embryonic kidney Fibroblast
    Known as: HEK293, 293

Method

pcDNA3.1-CiMetrnl-a or pcDNA3.1 (control) plasmid was transfected into the HEK293 cells using jetOPTIMUS reagent (Polyplus, France). Thirty-eight hours after transfection, the protein secretion inhibitor Brefeldin A (Beyotime, China) was added and the cells were cultured for 10 h The radio immunoprecipitation assay (RIPA, Beyotime, China) lysis containing phenylmethanesulfonyl fluoride (PMSF, Beyotime, China) was used to lyse the cells at 4 °C. Soluble protein was subjected to SDS-PAGE analysis for screening of mAb clones.

Abstract

Meteorin-like (Metrnl) is a novel immune regulatory factor or adipokine which is mainly produced by activated macrophages. In teleost fish, two homologs are present. In this study, monoclonal antibodies were prepared against recombinant grass carp (Ctenopharyngodon idella, Ci) Metrnl-a in mice and characterized by Western blotting, flow cytometry and immunofluorescent microscopy. In grass carp infected with Aeromonus hydrophila (A. hydrophila), the cells expressing CiMetrnl-a markedly increased in the gills, head kidney and intestine. In the inflamed intestine caused by A. hydrophila infection, the CiMetrnl-a producing cells were detected mainly in the mucosal layer of anterior, middle and posterior segments. Consistently, qRT-PCR analysis showed that the mRNA expression of CiMetrnl-a was markedly induced. Our results suggest that CiMetrnl-a is involved in regulating intestine inflammation caused by bacterial infection.

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