Identification and expression analysis of Langerhans cells marker Langerin/CD207 in grasscarp, Ctenopharyngodon idella.

Abstract

Langerhans cells (LCs) play an essential role in the initiation of immune response and maintenance of immune tolerance. However, the function and the molecular markers of grass carp LCs remains unclear. The grass carp LCs were firstly identified by immunofluorescence (IF) using a commercial anti-human Langerin/CD207 polyclonal antibody (pAb) and transmissionelectronmicroscope (TEM) technology in this study. After that, a cDNA sequence that homology with human and mouse CD207 gene was obtained by the bBLASTn program in NCBI. The open reading frame (ORF) of the grass carp CD207 gene contains 903bp encoding 300 amino acids which consisted of a transmembrane domain, a coiled-coil domain and a CLECT domain. Furthermore, the result of quantitative real-time PCR (qRT-PCR) indicated that this gene was expressed in all tested tissues, and mainly expressed in immune organs such as the gill, trunk kidney, head kidney, spleen and skin. To explore the role of CD207 gene in the immune responses induced by bacteria, an immersed infection model of grass carp with Flavobacterium columnare was constructed, and the optimal infection dose was determined to be 1.0×108 CFU/mL. Moreover, the qRT-PCR results indicated that the expression levels of CD207 gene were significantly upregulated at 6h, 12h, 1 d, 3 d and 7 d in the spleen, and significantly downregulated at 5 d in the head kidney, at 12h and 5 d in the gill, and at all time points in the skin after F. columnare infection. This result suggested that the grass carp CD207 gene may play an important role in antigen processing and presentation. Our results in this study suggested that CD207 gene is also existed in teleosts, and this study provided a molecular basis to analyzed the biological function of grass carp CD207 gene and the critical roles of LCs in the immune responses induced by bacterial infections.