Identification and bioactivity of a granulocyte colony-stimulating factor a homologue from large yellow croaker (Larimichthys crocea). Issue 98 (March 2020)

• Record Type: Journal Article
• Authors: Li, Qiuhua; Chen, Yuhong; Xu, Libing; Yang, Yusheng; Wen, Qiao; Gu, Li; Ao, Jingqun; Chen, Xinhua
• Is Part Of: Fish & shellfish immunology. Issue 98(2020)
• Page Start: 167
• Page End: 175
• Publication Date: 2020-03
• Subjects: Granulocyte colony-stimulating factor a (GCSFa) -- Large yellow croaker (Larimichthys crocea) -- Proliferation -- Signal pathway -- STAT3 and STAT5; Fishes -- Immunology -- Periodicals; Shellfish -- Immunology -- Periodicals; Poissons -- Immunologie -- Périodiques; Crustacés -- Immunologie -- Périodiques; 571.9617
• DOI: 10.1016/j.fsi.2020.01.003
• Languages: English
• ISSNs: 1050-4648

Abstract

Abstract: Granulocyte colony-stimulating factor (GCSF) is a growth factor that drives the proliferation and differentiation of granulocytes and monocytes/macrophages. Currently, two copies of GCSF, named GCSFa and GCSFb, have been identified in teleost fish, but data on the functions and signal pathways of these fish GCSFs are still limited. In the present study, a GCSFa homologue ( Lc GCSFa) was identified from large yellow croaker ( Larimichthys crocea ). The open reading frame (ORF) of Lc GCSFa is 636 bp long and encodes a protein of 211 amino acids (aa), with a 19-aa signal peptide and a typical IL-6 domain, conserved in fish GCSF sequences. The phylogenetic analysis showed that Lc GCSFa clustered with other fish GCSFa homologues. Lc GCSFa was constitutively expressed in all tissues tested and significantly up-regulated in head kidney and spleen by Vibrio alginolyticus or poly(I:C). Lc GCSFa transcripts were also detected in primary head kidney leucocytes (PKL), primary head kidney macrophages (PKM), and primary head kidney granulocytes (PKG), and significantly up-regulated in PKL and PKG by LPS or poly(I:C) . These data indicated that Lc GCSFa may be involved in the immune responses induced by bacterium and virus. The recombinant Lc GCSFa protein (r Lc GCSFa) produced in Pichia pastoris promoted the proliferation of PKL both in vivo and in vitro . Furthermore, r Lc GCSFa significantly increased both transcription and phosphorylation levels of the signal transducers and activators of transcription (STAT) proteins ( Lc STAT3 and Lc STAT5) in PKL, which are required for the GCSF-dependent proliferation. These results showed that Lc GCSFa may promote the proliferation of PKL via the activation of Lc STAT3 and Lc STAT5, suggesting a conserved role across vertebrate GCSFs. Highlights: A GCSFa homologue ( Lc GCSFa) was identified from large yellow croaker. Lc GCSFa transcripts were significantly up-regulated after Vibrio alginolyticus, LPS or poly(I:C) stimulation. Recombinant Lc GCSFa induced the proliferation of PKL both in vitro and in vivo. Recombinant Lc GCSFa significantly increased both transcription and phosphorylation levels of Lc STAT3 and Lc STAT5 in PKL. Lc GCSFa may promote the proliferation of PKL via the activation of Lc STAT3 and Lc STAT5.