House Dust Mite-induced Expression Of TIM4in Dendritic Cells Promotes To Th2Polarization

ObjectiveTo study expression of TIM4in mature dendritic cells from murine bone marrow that stimulated by HDM in vitro? and promotes native T cells differentiate into Th2cells.Methods1. To isolate and culture the murine bone marrow-derived mature dendritic cells (mDCs), the morphological changes of culture cells were observed under inverted microscope every day, CDllc、 CD80、MHC-II and CD86molecules were detected by flow cytometry.2. Bone marrow-derived mDCs were divided into four groups:control group (0U/ml HDM),100U/ml HDM group,200U/ml HDM group and400U/ml HDM group. mDCs were cultured with different groups for48h. The expression of TIM4was measured by flow cytometry and Western Blotting respectively.3. Different sets mDCs that including control group and400U/ml HDM group respectively were co-cultured with native T cells for48h. Level of interleukin-4(IL-4) and interferon-γ (IFN-γ) in culture medium were evaluated by enzymelinked immunosorbent(ELISA).Results1. The DCs were gathered at the tenth day. CDllc、CD80、MHC-Hand CD86molecules were detected by flow cytometry,and the results were (95.18±1.20)%,(85.07±2.05)%,(90.85±1.24)%and (70.78±1.57)%respectively. The results showed the cells were mDCs.2. After being co-cultured with HDM with different concentration, TIM4in mDCs was measured by flow cytometry and Western Blotting respectively. Flow cytometry showed the percentage of TIM4±DCs in different groups were (5.01±3.02)%,(12.78±1.57)%,(24.17±2.45)%and (55.85±2.24)%respectively, the expression of TIM4were increased in DCs compared with control group(P<0.05). And the Western Blotting also showed TIM4were increased in DCs compared with control group(P<0.05).3. The level of IL-4and IFN-γ in control group showed significant differences with400U/mlHDM group. Level of IL-4were (2.52±2.80) pg/mland (43.10±5.20) pg/mlin control group and400U/ml HDM group respectively; and level of IFN-γwere (90.37±3.74) pg/mland (38.74±3.67) pg/ml.ConclusionIn conclusion, the expression of TIM4in murine bone marrow-derived mDCs was significantly increased in response to HDM in a dose-dependent manner. TIM4+DCs promote native T cells differentiate into Th2cells.