可溶性抗CD47单抗对人树突细胞分化与功能的影响

Effect of soluble anti-CD47 monoclonal antibody on the differentiation and function of human dendritic cells

目的探讨抗CD4 7可溶性单抗B6H12对树突细胞分化及功能的影响。方法利用rhGM CSF、IL 4、细菌脂多糖 (LPS)组合在体外诱导人外周血单核细胞为树突细胞 ,并在培养体系中添加单抗B6H12。以透射电镜观察树突细胞形态 ,流式细胞术分析树突细胞膜表面分子表达。半定量RT PCR法及ELISA法分别检测树突细胞IL 12mRNA表达水平及蛋白质含量。Brdu ELISA法检测树突细胞刺激同种异型淋巴细胞增殖能力。结果树突细胞膜表面有CD4 7高表达 (94 %～ 98% )。B6H12单抗处理的树突细胞组与未加B6H12单抗组比较 ,CD80 + 细胞为 (6 8.14± 7.4 1) %vs(89.17± 8.5 9) % ;CD86 + 细胞为 (6 7.33± 4 .71) %vs(87.2 7± 3.5 6 ) % ;CD83+ 细胞为 (40 .0 8± 14 .80 ) %vs(72 .77± 8.6 8) % ;CD1a+ 细胞为 (6 6 .4 5± 4 .0 6 ) %vs(95 .93± 3.0 3) % ,HLA DR+ 细胞为 (40 .6 7± 13.4 8) %vs(98.97± 1.0 1) %。B6H12单抗显著地下调树突细胞IL 12mRNA及IL 12 P70 表达水平 (P <0 .0 5 ) ,且Brdu掺入量也显著的降低 (P <0 .0 1)。结论 可溶性CD4 7单抗能影响人树突细胞向成熟分化 ,并抑制其功能。

ObjectiveTo explore the influences of anti CD47 monoclonal antibody (mAb) B6H12 on the differentiation and function of cultured dendritic cells(DCs). MethodsHuman peripheral monocyte derived DCs were propagated with granulocyte macrophage colony stimulating factor(GM CSF) and lipopolysaccharide(LPS) plus interleukin 4(IL 4) in the presence or absence of soluble B6H12. Flow cytometry was used to analyze the immunophenotypes of cells, semi quantitative RT PCR and ELISA methods to analyse the mRNA and protein expression levels of interleukin 12(IL 12). The antigen presenting function of DCs was determined in one way mixed leukocyte reaction by Brdu ELISA technique.ResultsThere was a high expression rate(94%～98%) of CD47 molecules in DCs . The cell immunophenotypes in B6H12 mAb treated and untreated DC groups were as follows :CD80 + (68.14±7.41)%vs (89.17± 8.59)%; CD86 + ( 67.33 ±4.71)% vs (87.27±3.56)%; CD83(40.08±14.80)% vs (72.77± 8.68)%; CD1a + (66.45 ± 4.06 )% vs (95.93±3.03)%; and HLA DR(40.67±13.48) vs (98.97±1.01)%, respectively. The expression levels of mRNA and protein of IL 12 were strongly inhibited in B6H12 mAb treated DC(P< 0.01 ).The quantity of Brdu was also lower in B6H12 mAb treated DC than that in untreated DCs (P< 0.01 ).ConclusionThe anti CD47 McAb exerts a negative effect on the maturation and functions of cultured DCs.