Identification and functional characterization of CD154 in T cell-dependent immune response in Nile tilapia (Oreochromis niloticus). Issue 111 (April 2021)

• Record Type: Journal Article
• Authors: Li, Bingxi; Li, Yuan; Wu, Siwei; Yang, Yanjian; Fu, Shengli; Yin, Xiaoxue; Tu, Xiao; Fang, Liang; Guo, Zheng; Ye, Jianmin
• Is Part Of: Fish & shellfish immunology. Issue 111(2021)
• Page Start: 102
• Page End: 110
• Publication Date: 2021-04
• Subjects: Nile tilapia -- Oreochromis niloticus -- CD154 -- T cell-dependent immune response -- KLH; Fishes -- Immunology -- Periodicals; Shellfish -- Immunology -- Periodicals; Poissons -- Immunologie -- Périodiques; Crustacés -- Immunologie -- Périodiques; 571.9617
• DOI: 10.1016/j.fsi.2021.01.009
• Languages: English
• ISSNs: 1050-4648

Abstract

Abstract: CD154, a member of the TNF superfamily, is a multifunctional molecule highly expressed in activated T cells, and plays important roles in T cell-dependent humoral immune response. In this study, CD154 of Nile tilapia ( Oreochromis niloticus ) was identified, and its functions in the T cell-dependent immune response were demonstrated. The open reading frame (ORF) of OnCD154 is 699 bp, encoding a protein of 232 amino acids with a 23 amino acid transmembrane region. Amino acid sequence of OnCD154 is highly homologous to that of other teleost fish, especially rainbow trout. Quantitative real-time PCR (qRT-PCR) demonstrated that mRNA of OnCD154 is highly expressed in immune organs, especially in spleen, thymus, gills, head kidney, etc. In addition, the anti-OnCD154 polyclonal antibody (anti-(r)OnCD154) was successfully prepared, and it can react with natural protein in head kidney leukocytes. Following two immunizations with keyhole limpet hemocyanin (KLH) in vivo, the significantly up-regulated expression level of OnCD1 54 mRNA appeared earlier (fifth day) and higher (42.9 folds) in the second challenge than the first on in head kidney. Further, after stimulation with KLH in vitro, the expressions of T cell-dependent immune response-related molecules (activated T cell specific surface molecules CD3ε and CD154 ) and B cell differentiation-related molecules ( Blimp1 and sIgM ) and CD40 were significantly up-regulated in head kidney leukocytes. Moreover, the up-regulated expressions of these molecules were blocked with the treatment of anti-(r)OnCD154 antibody. Taken together, these results indicate that OnCD154 might get involved in T cell-dependent immune response, and provide a new insight into the humoral immune response of teleost fish. Highlights: The CD154 gene was identified in Nile tilapia ( OnCD154 ). OnCD154 expression was significantly up-regulated following KLH challenge ( in vivo and in vitro ). The significant up-regulation of OnCD154 was observed higher and earlier after the second stimulation than the first one. T cell-dependent immune response was inhibited by the treatment of anti-OnCD154 Ab in vitro.