Galectin-1Induced Bone Marrow Derived Dendritic Cells In Prevention And Treatment Of GVHD

Objective:To study the changes including the expression of life cycle, BM-DC surface molecules and cytokine secretion, as well as the differentiation, apoptosis and cytokines secretory effect of CD4+T cells treated with Galectin-1on bone marrow derived dendritic cells(BM-DC), And to construct graft-vesus-host disease(GVHD) mouse model to demenstrate whether BM-DCgal could effectively prevet GVHD.Methods:(1) Bone marrow progenitor cells were prepared from BALB/c mice and intruduced by GM-CSF and IL-4to generate DC, while adding different doses of Galectin-1, then the cells were cultured and to be taken photos for cell morphology and colony changes through microscope on Day0,1,3,5,7.(2) Detected Annexin V+7-AAD of Galectin-1processed cells by flow cytometry, also the expression of surface molecules as CD40, CD43, CD45RB, CD80, CD11c.(3) Cytokine secretion, including IL-2, IL-4, IL-6, IL-10, IL-27and TGF-beta, was detected by ELISA.(4) Dendritic cells were sorted by flow cytometry.(5) Spleen lymphocytes were prepared from C57BL/6mice.(6) Naive CD4+T cells were sorted by MACS.(7) Naive CD4+T cells were co-cultured with DCs sorted by flow cytometry, and the pictures were taken with microscope for morphology on DayO, Day1, Day3, Day5.(8) To detect the expression of IL-10, IL-17A, IFN-gama of BM-DC/BM-DCgal processed CD4+T cells by flow cytometry. (9) Cytokines secreted from BM-DC/BM-DCgal processed CD4+T cells, including IL-4, IL-10, IL17A, IFN-gama, were detected by ELISA.(10) The expression of IL-27receptor on processed CD4+T cell suface was detected by quantitative real-time PCR.(11) GVHD models were built by injecting bone marrow/spleen cells of C57BL/6mice to gamma ray irradiated BALB/c mice.Results:(1) Successfully prepared and cultured the dendritic cells.(2) The surface molecules on dendritic cells changed through Galectin-1treatment, also the life cycle was impacted.(3) Galectin-1treatment changed the cytokines secretion of DC.(4) The differentiation and survival period of naive CD4+T cells could be changed by co-culturing with Galectin-1processed DC.(5) The cytokines secretion of CD4+T cells could be changed by co-culturing with Galectin-1processed DC.(6) It showed that expression of IL-27receptor on CD4+T cells which co-cultured with Galectin-1processed DC could be upregulated.Conclusions:This study showed that naive CD4+T cells could be induced by Galectin-1processed DC and differentiated into Trl cells, which may provide experimental basis for the downregulation of T cell immune response in GVHD.