Cellular activities during a mixed leucocyte reaction in the teleost sea bass Dicentrarchus labrax
| Autor: | Giuseppe Scapigliati, Francesco Buonocore, Gianpaolo Zarletti, S. Meloni, Elisa Randelli, Stefania Benedetti |
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| Rok vydání: | 2005 |
| Předmět: |
T cell
T-Lymphocytes chemical and pharmacologic phenomena Aquatic Science Biology Immunofluorescence Flow cytometry Cell Line Adenosine Triphosphate Cyclosporin a medicine Leukocytes Environmental Chemistry Cytotoxic T cell Animals Sea bass Cytotoxicity Fluorescent Antibody Technique Indirect Analysis of Variance B-Lymphocytes medicine.diagnostic_test Reverse Transcriptase Polymerase Chain Reaction hemic and immune systems General Medicine Cytotoxicity Tests Immunologic Flow Cytometry Molecular biology In vitro medicine.anatomical_structure Immunology Cyclosporine Bass Lymphocyte Culture Test Mixed Immunosuppressive Agents |
| Zdroj: | Fishshellfish immunology. 20(5) |
| ISSN: | 1050-4648 |
| Popis: | In this investigation a number of “in vitro” activities of sea bass peripheral blood leucocytes (PBL) against allogeneic PBL inactivated by irradiation were studied. Stimulator PBL were cultured with inactivated allogeneic PBL, and direct counting of lymphocytes was done after 2 weeks by immunofluorescence and flow cytometry using mAbs DLT15 and DLIg3 specific for T-cells and B-cells, respectively. In a one-way mixed leucocyte reaction (MLR), results showed an increase of T lymphocytes, whereas B lymphocytes had values similar to those in control PBL. The increase of T-cells in MLR cultures was also confirmed using RT-PCR by analyzing the expression of the T-cell receptor (β-subunit) mRNA. The addition of 5 μg/ml of cyclosporin A (CsA) to the MLR caused a significant decrease in T-cell proliferation. Leucocytes from MLR cultures displayed an enhanced cytotoxic activity against xenogeneic target cells with respect to control PBL, raising the possibility of the presence of cytotoxic-like T lymphocytes. Cellular activation of PBL was confirmed in 2 weeks MLR by measuring antibody-induced intracellular Ca ++ mobilization with Fura-2 AM. This work represents the first direct quantitative determination of an “in vitro” T-cell activity in a teleost species. |
| Databáze: | OpenAIRE |
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