Effect of Immunogenicity of Neuroblastoma Cell Lines by Transduction of Adenovirus-Mediated Co-Stimulated Factor B7-1 and Granulocyte-Macrophage Colony-Stimulating Factor Genes
- VernacularTitle:腺病毒介导共刺激分子B7-1和粒细胞-巨噬细胞集落刺激因子基因对神经母细胞瘤细胞免疫原性的影响
- Author:
fang, CHEN
;
lei, LIU
- Publication Type:Journal Article
- Keywords:
neuroblastoma;
immunotherapy;
adenovirus;
co-stimulated factor B7-1;
granulocyte-macrophage colony-stimulating factor
- From:Journal of Applied Clinical Pediatrics
1992;0(05):-
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo observe the effect of co-stimulated factor B7-1 and granulocyte-macrophage colony-stimulating factor(GM-CSF) genes mediated by recombinant adenovirus on immunogenicity of neuroblastoma(NB)cell line SH-SY5Y.MethodsAfter NB cell line SH-SY5Y was transfected with recombinant adenovirus mediating green fluorescent protein(Ad-GFP) in different multiplicity of infection(MOI),the transfer efficiency was tested by flow cytometry(FCM).SH-SY5Y cells were transfected separately with recombinant adenovirus,recombinant adenovirus mediating B7-1 and GM-CSF genes or only B7-1 gene,and they were named as Ad,B7-GM and B7 group.The cells which were not transfected were named as SH group.The protein expression of B7-1 and GM-CSF were detected by using FCM and enzyme linked immunosorbent assay(ELISA).Peripheral mononuclear cells were separated and purified with Ficoll density gradient centrifugation.Cpm and cytokine(IFN-? and IL-2) secretory were evaluated by mixed lymphocyte culturing with different ratio(5 to 1 and 10 to 1) on target cells and effect cells.ResultsTransfer efficiency of recombinant adenovirus to SH-SY5Y was high.After transfection,B7-1 and GM-CSF genes were expressed.Proliferation of lymphocytes was stimulated and cytokine increased vigorously in B7-GM group and B7 group,and the effect in B7-GM group was stronger than that in B7 group.And the effect in ratio 5 to 1 group was stronger than that in 10 to 1 group.ConclusionsB7-1 and GM-CSF genes have synergistic effect.After transfection with recombinant adenovirus mediated B7-1 and GM-CSF genes,the immunogenicity of SH-SY5Y is greatly enhanced.
