The Maturation And Function Study Of Lentinan On Dendritic Cells Of Acute Myeloid Leukemia Patients In Vitro And In Vivo

Objective To explort the regulatory effects of lentinan (LNT) on maturation and function of dendritic cells (DC) of acute myeloid leukemia patients in vitro and in vivo, further clarify the immunological activity of lentinan, and to explore new ways of leukemia immunotherapy.Method Bone marrow mononuclear cells (BMNCs) from acute myeloid leukemia complete remission (AML-CR) patients were isolated with Ficoll density gradient centrifugation method,and were cultured in RPMI-1640 medium containing recombinant human granulocyte colony stimulating factor (GM-CSF)/recombinant human interleukin 4 (IL-4) placed in 5%CO2air at 37℃incubator. After culture for 7 days,positive control group was affected by a conventional dose of lipopolysaccharide (LPS), experimental groups were stimulated by LNT at lug,3ug and 5ug dose,negative control group was treated by RPMI-1640 medium of an equal volume.After 48 hours of stimulation, Morphology of DCs was observed under inverted microscope and Wright-Giemsa staining respectively. Phenotypes on the cells were identified by flow cytometer.Cell populations were phenotyped with the following FITC-or PE-conjugated MoAbs:CD80, CD83, CD86, CD1a, HLA-DR. IL-12 production was detected by ELISA. Dendritic cells in complete remission acute myeloid leukemia patients were isolated from human peripheral blood mononuclear cells (PBMNC) by magnetic cell sorting(MACS),we analyzed the concentration of IL-12 secreted by DCs using ELISA.Results In vitro effect of high doses of LNT showed typical DC morphology,could increase the expression of CD80,CD83,CD86,CD1a and HLA-DR molecules on DC surface(P<0.05),LNT-treated DCs secreted higher level of IL-12 than negative control group(P<0.05),and was concentration-dependent Lentinan.LNT-treated DCs secreted significantly higher level of IL-12 than untreated DCs(P<0.05),consistent with in vivo and in vitro.Conclusion LNT could promote functional and phenotypic maturation of dendritic cells of acute myeloid leukemia patients in vivo and in vitro,LNT might be one of its anti-leukemia mechanism.