RESUMO
Scavenger receptors (SRs) are important pattern recognition receptors (PRRs), which play significant roles in host defense against pathogens by identifying pathogen-associated molecular patterns (PAMPs). In this study, we report the cloning and characterization of a SR from Eriocheir sinensis (EsSR-B1) which is a 500 amino acid protein encoded by a gene comprised of 2726 nucleotides with a 1503 bp open reading frame. The domains of EsSR-B1 were found to be evolutionarily conserved. EsSR-B1 was widely detected in different tissues of E. sinensis and significantly up-regulated in hemocytes after stimulation by Staphyloccocus aureus or Vibrio parahaemolyticus. Recombinant EsSR-B1 protein could bind to bacteria and promote phagocytosis upon bacterial stimulation. Moreover, antimicrobial peptide expression was reduced in EsSR-B1-silenced hemocytes after challenge by S. aureus or V. parahaemolyticus. Thus, EsSR-B1 has a critical role in the binding of bacteria and subsequent promotion of hemocyte phagocytosis.
Assuntos
Braquiúros/genética , Braquiúros/imunologia , Regulação da Expressão Gênica/imunologia , Imunidade Inata/genética , Receptores Depuradores/genética , Receptores Depuradores/imunologia , Sequência de Aminoácidos , Animais , Proteínas de Artrópodes/química , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/imunologia , Sequência de Bases , Feminino , Perfilação da Expressão Gênica , Hemócitos/metabolismo , Masculino , Fagocitose , Filogenia , Distribuição Aleatória , Receptores Depuradores/química , Alinhamento de Sequência , Staphylococcus aureus/fisiologia , Vibrio parahaemolyticus/fisiologiaRESUMO
Vitellogenin (Vtg) is traditionally regarded as a key supplier of nutrients and energy during the early development of embryos and larvae, but accumulating evidence suggests that Vtg is also involved in innate immune defense. Whether Vtg is involved in innate immunity in Eriocheir sinensis, and its functions, remain largely unknown. In this study, a cDNA representing the vitellogenin1 gene from E. sinensis (Es-vtg1) was cloned. The full-length Es-vtg1 cDNA comprised 7939 nucleotides, encoding an open-reading frame of 2567 amino acid residues. Bioinformatic analysis showed that the domains of Es-Vtg1 have been conserved during evolution. Quantitative real-time PCR and western blotting showed that Es-vtg1 was highly expressed in ovary and hepatopancreas. Moreover, bacteria could induce the high-level expression of Es-Vtg1. Es-Vtg1 plays important roles in immunological defense, including binding to bacteria, inhibiting bacterial proliferation, and regulating the expression of antimicrobial peptides. Collectively, these results demonstrate that Es-Vtg1 plays critical roles in antimicrobial function.
