Molecular characteristics and functional analysis of non-specific cytotoxic cell receptor (NCCRP1) in golden pompano (Trachinotus ovatus)

Non-specific cytotoxic cells (NCCs) are cytotoxic cell population found in innate immune system of teleost, playing crucial role in immune defense. Non-specific cytotoxic cell receptor protein 1 (NCCRP1) is responsible for recognizing target cells and activating NCCs. That said, since the studies regarding NCCs' role in fish during pathogen infection are few, it is necessary to conduct more comprehensive studies. In this study, we identified NCCRP1 from Trachinotus ovatus (ToNCCRP1). The open reading frame of ToNCCRP1 was found to be 702 bp, encoding a protein of 233 amino acids. Additionally, ToNCCRP1 contained a conserved F-box-associated domain and exhibited more than 61 % similarity to NCCRP1 in other fish species. Quantitative real-time PCR analysis showed that ToNCCRP1 mRNA was generally expressed in all tissues, with the highest level expressed in the liver. Furthermore, the expression of ToNCCRP1 was significantly upregulated following infection with Strep-tococcus iniae. In vitro experiments demonstrated that recombinant ToNCCRP1 possessed bacterial agglutination and binding capabilities, suggesting its antibacterial function. Additionally, we investigated the immune response of head kidney leukocytes (HKLs) to ToNCCRP1. The challenge experiments revealed that ToNCCRP1 played a role in the immune response by influencing the inflammatory response, regulating signaling pathways and apoptosis in HKLs. These findings suggest that NCCRP1 is involved in the immune defense against patho-genic infections in golden pompano, providing insights into the immune mechanisms of teleost.