Zhao-Hui Ma1, Hang Lu, Qiang Lu, Zhi-Fa Yao, Yong Han. 1. Department of Thoracic Surgery, Tangdu Hospital, Fourth Military Medical University, Xi'an, People's Republic of China.
Abstract
BACKGROUND: Dendritic cells (DCs) are the principal antigen-presenting cells involved in primary immune response and immunoregulation. The function of DCs is believed to depend on their degree of maturation. Mature DCs activate immune responses, whereas immature DCs (imDCs) tend to induce immune tolerance. CD1 is involved in regulating the development of imDCs, which have important roles in initiating or suppressing the immune response after transplantation. MATERIALS AND METHODS: We used male BALB/c mice and C57BL/6 mice (aged 8-10 wk, 18-22 g). We isolated and purified T lymphocytes from mouse spleen. Immature DCs modified by viral delivery of interleukin-10 (IL-10) were stimulated with granulocyte macrophage colony-stimulating factor and lipopolysaccharide (LPS) and treated with anti-CD1d in vitro. We used mixed lymphocyte cultures to evaluate the heterogeneity of T lymphocyte response. We also examined the proliferation of T lymphocytes and the expression of cytokines. RESULTS: CD1d blockade did not impair granulocyte macrophage colony-stimulating factor and LPS-stimulated DC maturation. We observed a dramatic increase in allogeneic T lymphocyte proliferation (stimulation index) at all tested responder-stimulator ratios in response to imDCs cultured in the presence of LPS (P < 0.05). CD1d has an important role in imDC-primed T cell response (P < 0.05). CD1d blockade reduced the capacity of imDCs to prime allogeneic T cells. T cells pre-sensitized by LPS-stimulated imDCs showed remarkably elevated proliferation in response to T cells from either BALB/c or C57BL/6 mice (P < 0.01). We observed a significant decrease in the proliferation of T cells pre-sensitized by stimulated imDCs after CD1d blockade. Lipopolysaccharide stimulation caused elevated the production of IL-12 and tumor necrosis factor-α (TNF-α) (P < 0.01) and decreased the secretion of IL-10 (P < 0.05). The addition of CD1d neutralization antibody did not significantly change the concentrations of IL-12, TNF-α, or IL-10 produced by imDCs cultured in the presence of LPS (P > 0.05). CONCLUSIONS: Blockade of CD1d impaired the ability of imDCs to stimulate allogeneic T cell response. By reduced T cell proliferation, the secretion of IL-12 and TNF-α decreased and production of a T-helper type 2 cytokine IL-10 increased, which indicates the potential of CD1d blockade as a method to induce immune tolerance to allograft antigens in transplantation.
BACKGROUND: Dendritic cells (DCs) are the principal antigen-presenting cells involved in primary immune response and immunoregulation. The function of DCs is believed to depend on their degree of maturation. Mature DCs activate immune responses, whereas immature DCs (imDCs) tend to induce immune tolerance. CD1 is involved in regulating the development of imDCs, which have important roles in initiating or suppressing the immune response after transplantation. MATERIALS AND METHODS: We used male BALB/c mice and C57BL/6 mice (aged 8-10 wk, 18-22 g). We isolated and purified T lymphocytes from mouse spleen. Immature DCs modified by viral delivery of interleukin-10 (IL-10) were stimulated with granulocyte macrophage colony-stimulating factor and lipopolysaccharide (LPS) and treated with anti-CD1d in vitro. We used mixed lymphocyte cultures to evaluate the heterogeneity of T lymphocyte response. We also examined the proliferation of T lymphocytes and the expression of cytokines. RESULTS:CD1d blockade did not impair granulocyte macrophage colony-stimulating factor and LPS-stimulated DC maturation. We observed a dramatic increase in allogeneic T lymphocyte proliferation (stimulation index) at all tested responder-stimulator ratios in response to imDCs cultured in the presence of LPS (P < 0.05). CD1d has an important role in imDC-primed T cell response (P < 0.05). CD1d blockade reduced the capacity of imDCs to prime allogeneic T cells. T cells pre-sensitized by LPS-stimulated imDCs showed remarkably elevated proliferation in response to T cells from either BALB/c or C57BL/6 mice (P < 0.01). We observed a significant decrease in the proliferation of T cells pre-sensitized by stimulated imDCs after CD1d blockade. Lipopolysaccharide stimulation caused elevated the production of IL-12 and tumor necrosis factor-α (TNF-α) (P < 0.01) and decreased the secretion of IL-10 (P < 0.05). The addition of CD1d neutralization antibody did not significantly change the concentrations of IL-12, TNF-α, or IL-10 produced by imDCs cultured in the presence of LPS (P > 0.05). CONCLUSIONS: Blockade of CD1d impaired the ability of imDCs to stimulate allogeneic T cell response. By reduced T cell proliferation, the secretion of IL-12 and TNF-α decreased and production of a T-helper type 2 cytokine IL-10 increased, which indicates the potential of CD1d blockade as a method to induce immune tolerance to allograft antigens in transplantation.