title: Characterizations of CD4-1, CD4-2 and CD8β T cell subpopulations in peripheral blood leucocytes, spleen and head kidney of Japanese flounder (Paralichthys olivaceus).
abstract: In the previous study, antibodies against CD3 molecule have been produced and were used in labeling T cells in Japanese flounder (Paralichthys olivaceus). In this paper, CD4+and CD8+lymphocytes subpopulations in peripheral blood leucocytes (PBL), spleen and head kidney of flounder were investigated. The flounder CD4-1, CD4-2 and CD8β recombinant proteins and their antibodies (Abs) were produced, then the cross-reactivity of the Abs to CD4-1, CD4-2 and CD8β was detected by Western blotting, respectively, and the reactions of Abs to PBL were analyzed by immunofluorescence staining (IFS) and flow cytometry (FCM). CD4-1+/CD3+, CD4-2+/CD3+, and CD8β+/CD3+lymphocytes in PBL, spleen and head kidney were observed by double IFS, then their proportions were analyzed using two-color FCM, respectively. Further, CD4-1/CD8β, CD4-2/CD8β, or CD4-1/CD4-2 lymphocytes were analyzed using double-IFS and two-color FCM. Finally, CD4-1+, CD4-2+, and CD8β+lymphocytes in spleen and head kidney were observed by immunohistochemistry. The results showed that the Abs were specific for CD4-1, CD4-2 and CD8β molecules, respectively. The proportions of CD4-1+/CD3+, CD4-2+/CD3+, and CD8β+/CD3+lymphocytes were 6.7±2.0%, 8.6±2.8%, 2.1±1.3% in PBL; 13.6±3.6%, 15.6±5.2%, 2.8±1.4% in spleen; 20.0±4.6%, 20.5±4.6%, 3.2±1.5% in head kidney, respectively. Most CD4+and CD8+cell subpopulations belonged to CD3+cells; there were no cross-reactivity between CD4+and CD8+cells. CD4-1+/CD4-2-, CD4-1-/CD4-2+, and CD4-1+/CD4-2+cells presented different proportions in PBL, spleen and head kidney, among them, CD4-1+/CD4-2+cell is the majority of CD4T cell subpopulation.
authors.affiliation: Laboratory of Pathology and Immunology of Aquatic Animals,KLMME, Ocean University of China, Qingdao 266003, PR China.
authors.forename: Jing
authors.initials: J
authors.lastname: Xing
authors.affiliation: Laboratory of Pathology and Immunology of Aquatic Animals,KLMME, Ocean University of China, Qingdao 266003, PR China.
authors.forename: Junjie
authors.initials: J
authors.lastname: Ma
authors.affiliation: Laboratory of Pathology and Immunology of Aquatic Animals,KLMME, Ocean University of China, Qingdao 266003, PR China.
authors.forename: Xiaoqian
authors.initials: X
authors.lastname: Tang
authors.affiliation: Laboratory of Pathology and Immunology of Aquatic Animals,KLMME, Ocean University of China, Qingdao 266003, PR China.
authors.forename: Xiuzhen
authors.initials: X
authors.lastname: Sheng
authors.affiliation: Laboratory of Pathology and Immunology of Aquatic Animals,KLMME, Ocean University of China, Qingdao 266003, PR China; Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, No. 1 Wenhai Road, Aoshanwei Town, Qingdao, PR China. Electronic address: wbzhan@ouc.edu.cn.
authors.forename: Wenbin
authors.initials: W
authors.lastname: Zhan
mesh_terms: D000818:Animals; D015153:Blotting, Western; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000166:cytology / Q000276:immunology; D005432:Flounder / Q000276:immunology*; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D060226:Head Kidney / Q000166:cytology / Q000276:immunology*; D007962:Leukocytes / Q000166:cytology / Q000276:immunology*; D013154:Spleen / Q000166:cytology / Q000276:immunology*; D016176:T-Lymphocyte Subsets / Q000276:immunology*
keywords: CD3; CD4; CD8; Paralichthys olivaceus; T cell