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  • Title: [Influence of different cyto-stimuli on immunological activity of splenic regulatory T cells in mice].
    Author: Zhang Y, Yao YM, Chang Q, Dong N.
    Journal: Zhongguo Wei Zhong Bing Ji Jiu Yi Xue; 2007 Mar; 19(3):142-5. PubMed ID: 17376265.
    Abstract:
    OBJECTIVE: To investigate the expression of cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) and intranuclear forkhead/winged helix transcription factor p3 (Foxp3) of splenic regulatory T cells (Tregs) in mice, and compare the effects of different stimuli on the immunological activity of Tregs. METHODS: CD4+CD25+ Tregs isolated from the spleens of male BALB/c mice by magnetic beads were seeded on 96-well (1x10(5) cells/well) cell culture plates supplemented with phytohemagglutin (PHA, 20 mg/L), concanavalin A (ConA, 5 g/L) or coated with anti-CD3 (1 mg/L). After being stimulated with different stimuli for various length of time, the expressions of CTLA-4 and Foxp3 molecules of Tregs were determined. The time-dependent responses between different stimuli and CTLA-4 as well as Foxp3 were analyzed by means of flow cytometry. RESULTS: After stimulation with PHA, the CTLA-4 expressions on surface of mice splenic Tregs and intranuclear Foxp3 molecules were unchanged (both P>0.05). In ConA-treated group, the expression of CTLA-4 was significantly up-regulated at 24 hours (P<0.01), while the CTLA-4 expression returned to the control range at 48 and 72 hours. The expression of Foxp3 was not markedly different between ConA-treated group and controls. When treated with plate-bound anti-CD3, both CTLA-4 and Foxp3 expressions were significantly up-regulated at 24, 48 and 72 hours (P<0.05 or P<0.01), especially at 24 and 48 hours (all P<0.01). CONCLUSION: These data indicated that various stimuli can result in different modulatory responses of the CTLA-4 and Foxp3 expressions in mice splenic Tregs. Anti-CD3 appears to be an effective immunoregulatory signal that influences the activation of CD4+CD25+ Tregs.
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