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.
journal_name
Mol Immunoljournal_title
Molecular immunologyauthors
Xing J,Ma J,Tang X,Sheng X,Zhan Wdoi
10.1016/j.molimm.2017.02.015subject
Has Abstractpub_date
2017-05-01 00:00:00pages
155-165eissn
0161-5890issn
1872-9142pii
S0161-5890(17)30066-4journal_volume
85pub_type
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