A Primary Study On The Mechanisms Of Doxycyline And Salbutamol Regulating The Immune Function Of Rat Bone Marrow-Derived Dendritic Cells

Objective To explore the mechanisms of doxycycline (Dox) regulating the immune function of rat bone marrow-derived dendritic cells(RBM-DCs).Methods SD rat bone-marrow cells were isolated from rat femur under the asepsis condition. The haemopoietic stem cells were induced by recombinated rat granulocyte-macrophage colony-stimulating factor (rrGM-CSF) 4ng/ml, recombinated rat interleukin-4 (rrIL-4) 4ng/ml combined with Dox 100ng/ml, 500ng/ml or 1000ng/ml respectively for 8 days in vitro. A control was set and induced only by rrGM-CSF 4ng/ml + rrIL-4 4ng/ml simultaneously. The morphology of DCs was monitored by inversion microscope and the cell surface markers on dendritic cells were detected by flow cytometer. The regulatory effects of Dox on the immunostimulatory capacity of the DCs were evaluated by allogeneic mixed leukocytes reaction (MLR) between DCs and T cells.Results Compared with the control group, DCs morphology hadn t changed significantly after induced by rrGM-CSF +rrIL-4 + Dox. Different dose of Dox could downregulate the expression of CD11c , OX62, CD86, CD80 and MHC classâ…¡on DCs. The Dox-treated DCs inhibited the T cells proliferation significantly in MLR. Conclusions Dox could inhibit the differentiation of RBM-DCs and downregulate the antigen presenting function via decreasing the expression of their surface markers, including CD11c, OX62, CD86, CD80 and MHC classâ…¡. Objective To explore the mechanisms of salbutamol (Sal) regulating the immune function of RBM-DCs.Methods SD rat bone-marrow cells were isolated from rat femur under the asepsis condition. The haemopoietic stem cells were induced by rrGM-CSF 4ng/ml, rrIL-4 4ng/ml combined with Sal 250ng/ml, But 300ng/ml, Sal 250ng/ml + But 300ng/ml respectively for 8 days in vitro. A control was set and induced only by rrGM-CSF 4ng/ml + rrIL·-4 4ng/ml simultaneously. The morphology of DCs was monitored by inversion microscope. The cell surface markers on dendritic cells were detected by flow cytometer. The regulatory effects of Sal, But and Sal + But on the capacity of the immunostimulatory of DCs were evaluated by allogeneic mixed leukocytes reaction (MLR) between DCs and T cells.Results Compared with the control group, DCs morphology hadn't changed significantly after induced by rrGM-CSF + rrIL-4 + Sal 250ng/ml, But 300ng/ml, Sal 250ng/ml+ But 300ng/ml. Sal 250ng/ml could upregulate the expression of OX62 and downregulate CD86, CD80 and MHC classâ…¡on DCs, but there were no obvious effects on CD11c. But 300ng/ml could downregulate the expression of CD11c, CD86, CD80 and MHC classâ…¡on DCs also, but couldn't change the expression of OX62 significantly. Sal 250ng/ml +But 300ng/ml could downregulate the expression of CD11c, CD86, CD80 and MHC classâ…¡on DCs, however, could upregulate the expression of OX62．Both Sal 250ng/ml-treated DCs and But 300ng/ml-treated DCs could weaken the T cells proliferation dramatically in MLR. Sal 250ng/ml+But 300ng/ml-treated DCs could also inhibit the T cells proliferation in MLR. It meant that But couldn't reverse the effects of Sal.Conclusions Sal could downregulate the antigen presenting function of RBM-DCs via decreasing the expression of MHC classâ…¡and costimulatory molecules, but could promote the differentiation of RBM-DCs. But couldn't reverse the effects of Sal. So Sal and But might inhibit the immunoregulation function of RBM-DCs via other signal pathway, notβ₂-AR.