ObjectiveTo evaluate  the effect of recombinant bacillus CalmetteGuerin (BCG)rBCGAg85AESAT6（rBCGAE)on immunostimulatory activity of human macrophages, and  provide theoretical proof and experimental basis for  vaccine use.MethodsThe previously constructed rBCGAE and BCG strains were obtained respectively to infect the THP1 cells that stimulated by phorbol12myristate13acetate（PMA）. 24 h,48 h and 72 h after infection, expression of macrophage surface markers CD80 and CD86  were measured by flow cytometry, and the concentration of interferonγ (IFNγ) and tumor necrosis factorａ(TNFａ) in supernatant were detected with ELISA kit. ResultsAfter four hours culture, phagocytic rate of THP1  stimulated by PMA was  significantly higher than that nonstimulated by PMA （［91.26±2.13］% vs ［8.45±1.54］%，P=0.01）； 24 h,48 h and 72 h after infection, the percentages of CD86 positive cells (［32.84±7.13］%, ［48.42±5.46］%, ［39.48±5.67］%) and CD80 positive cells (［20.28±1.13%］, ［23.67±1.23］%, ［23.19±1.58］%) in rBCGAE group were obviously higher than those in BCG group (CD86：［28.17±5.26］%, ［40.09±7.21］%, ［31.26±6.85］%; CD80：［22.15±1.82］%, ［23.27±1.91］%, ［22.68±0.87］%, P<0.01). 24 h,48 h and 72 h after infection, the concentration of IFNγ (［1 986±156］pg/mL,［ 4 687±168］ pg/mL, ［3 238±97］ pg/mL) and TNFа(［1 153±48］ pg/mL, ［5 864±97］ pg/mL, ［4 129±68］pg/mL) in rBCGAE group were higher than those in BCG group (IFNγ：［1 245±32］pg/mL, ［3 067±143］ pg/mL, ［2 879±186］ pg/mL; TNFа：［486±18］ pg/mL, ［3 237±86］ pg/mL, ［1 068±74］ pg/mL, P<0.01).ConclusionRecombinant BCGAE can enhance immunostimulatory activity of human macrophages, and it is an improved TB vaccine as an alternative to BCG for further study.