C-reactive protein (CRP) and serum amyloid P component (SAP) are evolutionarily conserved soluble pattern-recognition molecules that play critical roles in innate immunity, but their functional divergence in teleosts remains poorly understood. Here we characterized the CRP and SAP homologs in spotted sea bass (Lateolabrax maculatus) and investigated their distinct immunomodulatory functions. The full-length cDNAs of CRP and SAP encode proteins of 230 and 224 amino acids, respectively, both containing typical pentraxin domains with conserved motifs. Both genes were constitutively expressed, with CRP predominant in the brain and SAP in the spleen. Following challenge with lipopolysaccharide (LPS), Poly (I:C), or Edwardsiella tarda, both genes were significantly upregulated in vivo and in vitro, confirming their identity as acute-phase proteins. Subcellular localization in spotted sea bass brain (LMB) cells and HEK-293T cells revealed distinct patterns: CRP was predominantly perinuclear, whereas SAP was enriched in the nucleus, suggesting non-canonical intracellular functions. Functional overexpression in LMB cells further demonstrated divergent roles: CRP suppressed TNFα while upregulating NF-κB1 and NF-κB2, whereas SAP specifically induced IL-4 and NF-κB1. These findings indicate that CRP and SAP have evolved distinct subcellular localization and regulatory functions, with CRP fine-tuning the NF-κB/TNFα axis and SAP promoting an IL-4-associated response. This study provides new insights into the functional diversification of pentraxins in teleost immunity.
Yushuai Xie, Ya-Wen Yang, Hai Yang et al.· Developmental and Comparativ...· 0 citations
T-cell receptor (TCR) signaling is essential for adaptive immunity in vertebrates, with the tyrosine kinases Fyn, Lck, and Zap-70 playing central roles in signal transduction. In the present study, the full cDNA sequences of Fyn, Lck, and Zap-70 from golden pompano (Trachinotus ovatus) were obtained by RT-PCR and RACE-PCR and their expression patterns in normal tissues and head kidney (HK) following lipopolysaccharide (LPS), polyinosinic-polycytidylic acid (polyI:C) stimulation and Vibrio alginolyticus infection were examined by qPCR. The open reading frames of trFyn, trLck, and trZap-70 respectively encoded 537, 513, and 609 amino acids, all containing conserved similar functional domains as their mammalian counterparts. qPCR analysis revealed that all three genes were ubiquitously expressed in examined tissues, with high expression in immune-related tissues. trFyn, trLck, and trZap-70 were significantly induced in HK after LPS, polyI:C stimulation and V. alginolyticus infection. Co-immunoprecipitation (co-IP) assay demonstrated that both trFyn and trLck could interact with trZap-70 in HEK-293T cells. These findings provide molecular basis for further study of the TCR signaling and T-cell mediated immunity in golden pompano.
You-Chuan Wei, Dongqiang Gan, Wen Qi et al.· Fish and Shellfish Immunolog...· 0 citations
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