1. Characterizations of CD4-1, CD4-2 and CD8β T cell subpopulations in peripheral blood leucocytes, spleen and head kidney of Japanese flounder (Paralichthys olivaceus).
- Author
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Xing, Jing, Ma, Junjie, Tang, Xiaoqian, Sheng, Xiuzhen, and Zhan, Wenbin
- Subjects
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CD4 antigen , *T cells , *CELL populations , *LEUCOCYTES , *PARALICHTHYS , *CD3 antigen - 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. [ABSTRACT FROM AUTHOR]
- Published
- 2017
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