The Grass Mycobacterium Fu36 Research On The Impact Of Childhood Acute Leukemia Dendritic Cells

PartⅠEffect of Mycobacterium phlei F.U.36 on the Culture and Proliferation of Dendritic Cells Derived from Children Acute Leucemia Peripheral Blood in vitroObjective To investigate the effect of Mycobacterium phlei F.U.36 suspended liquor (Utilin's,U) on the culture and proliferation of dendritic cells(DC) derived from children acute leucemia peripheral blood in vitro.Methods The mononuclear cells were isolated from children acute lymphoblastic leucemia peripheral blood and cultured with RPMI-1640 in the control group,while the test groups were cultured with U+RPMI-1640(U),rhGM-CSF+rhTNF-a+rhIL-4+ RPMI-1640(GIT),GIT+U+RPMI-1640(GITU) respectively.The growth of DCs was observed under a invert microscope everyday.We counted the numbers of different groups and detected the immunephenotypes of DCs by flow cytometry on the 8th day after cultured.Results The test groups all attained a certain amount of typical DCs;The numbers of cells and the rate of CD1a,HLA-DR,CD83 positive cells of the test groups are higher than those of the control group;But there is no difference between the numbers of cells and the rate of CD1a+ cells of U and GIT group;on the contrary,the the numbers of cells and the rate of CD1a~+,HLA-DR~+,CD83~+ cells of GITU group are higher than the other groups.Conclusion Mycobacterium phlei F.U.36 suspended liquor not only promotes the proliferation of DCs derived from children acute leucemia peripheral blood in vitro,but also cooperates with rhGM-CSF,rhTNF-a and rhIL-4 in proliferating and promoting the maturity of DCs. PartⅡEffect of Mycobacterium Phlei F.U.36 on the Immunifaction of Dendritic Cells Derived from Children Acute Leucemia Peripheral BloodObjective To investigate the effect of mycobacterium phlei F.U.36 suspended liquor on the immunifaction of Dendritic Cells Derivcd from Children Acute Leucemia Peripheral Blood.Methods The mononuclear cells were isolated from children acute lymphoblastic leucemia peripheral blood and cultured with RPMI-1640+rhGM-CSF+rhIL-4+rhTNF-a in the control group(GIT).The test groups contain four parts:U(uscd Utilin's only),GITU1-3(GIT combined with Utilin's in different concentration,which is put in 3rd day:GITU1,0.86ng/ml; GITU2,1.72ng/ml;GITU3,2.58ng/ml).The growth of DCs was observed under a invert microscope everyday and collected on the 8th day.we mixed the different groups's DCs with T cells,which just separated from another acute lymphoblastic leucemia peripheral blood. Cultured with rhIL-2+RPMI-1640 96 hours' later,we counted the numbers of different groups and test the contents of IL-12,IFN-γby ELISA.also,we mixed the different groups's DCs and T cells(Cultured with rhIL-2+RPMI-1640 for 5 days) and Culturcd for 72 hours. Cytotoxicity assay was measured by MTT method.Results All groups attained a certain amount of typical DCs,Mixcd lymphocyte reaction shows that with the increasing of DCs,the stimulation index of T cell is rising,also the contains of IL-12,IFN-γin supernatants is rising with the increasing of the concentration of Utilin's in test groups;Cytotoxicity assay shows that killing activity on HL-60 cells of T cells in test groups is highly than the control group,and in test groups,with the increasing of the concentration of Utilin's,the killing activity on HL-60 cells is dramaticly enhancing.Conclusion The DCs induced by Utilin's can particularly enhance the proliferation and secretion of T cells.With the increasing of the concentration of Utilin's,the function is enhancing,and the cytotoxicity on HL-60 cells of T cells is also increasing.It is more notable when combined with rhGM-CSF+rhTNF-a+rhIL-4.so,Utilin's can promote the maturity of DCs,reinforce its antigen presenting capability,accordingly enhance the anti-leukemic immune effect of DCs which derived from children acute leucemia peripheral blood.