title: Grouper (Epinephelus coioides) TCR signaling pathway was involved in response against Cryptocaryon irritans infection.
abstract: T cell activation is a complicated process accompanying with the activation of T cell receptor (TCR) signaling pathway, which is not well described in teleost fish. The initiation of this pathway depends on the interaction of membrane TCR co-receptors (e.g. CD4/8, CD3 and CD45) and a series of cytoplasmic protein tyrosine kinases (e.g. Lck, Fyn and ZAP70). Cyptocaryon irritans is a ciliate pathogen of marine fish white spot disease causing huge economic lost in marine aquaculture. This parasite can infect fish gill and skin and is considered to be a good pathogen model for fish gill and skin mucosal immunity. Our previous studies showed the locally mucosal antibody response was important for fish defense against this parasite. While how TCR signaling pathway involved in T cell activation to help B cell activation in C. irritans infected fish is still not known. In the present study, we cloned a grouper TCR co-receptor gene EcCD3ε (537 bp) and its three kinase genes, including EcLck (1512 bp), EcFyn (1605 bp) and EcZAP70 (1893 bp). Homology analysis showed that they all shared the highest identity with corresponding genes from Takifugu rubripes (EcCD3ε 41%, EcLck 88%, EcFyn 98% and EcZAP70 93%), and their conserved motifs involved in the signaling transduction were analyzed. The tissue distribution analysis showed these four genes were high expressed in thymus, and it is interesting to find their comparative high expression in skin, gill and midgut mucosal immune tissues. In C. irritans infected grouper, the expression of three TCR co-receptors (EcCD4-1, EcCD3ε and EcCD45) and three kinases (EcLck, EcFyn and EcZAP70) was tested in skin, gill, head kidney and spleen at 0, 12 h, 24 h, 2 d, 3 d, 5 d and 7 d. All six genes were significantly up-regulated in skin at most tested time points, which indicate the possibility of skin local T cell activation to support the local antibody response. Compared to three TCR co-receptors, significantly up-regulation of three kinases were seen in the spleen, and the spleen fold changes of these three kinases were much higher than head kidney, which indicates spleen maybe the major systematic immune organs for T cell activation in C. irritans infected fish.
authors.affiliation: School of Bioscience and Bioengineering, South China University of Technology, Guangzhou 510006, PR China.
authors.forename: Ze-Xiang
authors.initials: ZX
authors.lastname: Li
authors.affiliation: College of Marine Sciences, South China Agricultural University, Guangzhou 510642, Guangdong Province, PR China.
authors.forename: Yan-Wei
authors.initials: YW
authors.lastname: Li
authors.affiliation: School of Bioscience and Bioengineering, South China University of Technology, Guangzhou 510006, PR China.
authors.forename: Shun
authors.initials: S
authors.lastname: Xu
authors.affiliation: School of Bioscience and Bioengineering, South China University of Technology, Guangzhou 510006, PR China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Xu
authors.affiliation: College of Marine Sciences, South China Agricultural University, Guangzhou 510642, Guangdong Province, PR China.
authors.forename: Ze-Quan
authors.initials: ZQ
authors.lastname: Mo
authors.affiliation: College of Marine Sciences, South China Agricultural University, Guangzhou 510642, Guangdong Province, PR China. Electronic address: dxm72@scau.edu.cn.
authors.forename: Xue-Ming
authors.initials: XM
authors.lastname: Dan
authors.affiliation: School of Bioscience and Bioengineering, South China University of Technology, Guangzhou 510006, PR China. Electronic address: xcluo@scut.edu.cn.
authors.forename: Xiao-Chun
authors.initials: XC
authors.lastname: Luo
mesh_terms: D000818:Animals; D001492:Bass; D016798:Ciliophora / Q000502:physiology; D016770:Ciliophora Infections / Q000235:genetics / Q000276:immunology / Q000469:parasitology / Q000662:veterinary*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D005393:Fish Diseases / Q000235:genetics* / Q000276:immunology / Q000469:parasitology; D029941:Fish Proteins / Q000235:genetics* / Q000378:metabolism; D018928:Immunity, Mucosal; D010802:Phylogeny; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011948:Receptors, Antigen, T-Cell / Q000235:genetics* / Q000378:metabolism; D017422:Sequence Analysis, DNA / Q000662:veterinary; D015398:Signal Transduction
chemical_list: D018076:DNA, Complementary; D029941:Fish Proteins; D012333:RNA, Messenger; D011948:Receptors, Antigen, T-Cell
keywords: CD3ε; Cryptocaryon irritans; Epinephelus coioides; Fyn; Lck; ZAP70