title: [Response of mouse macrophage cell line infected with Bacille Calmette-Guerin in vitro].
abstract: OBJECTIVE
  To illuminate the response of mouse macrophage cell line RAW264.7 after infected with Bacille Calmette-Guerin (BCG) in vitro.
  
  
  METHODS
  We analyzed the morphology and expression of co-stimulatory molecules on the surface of RAW264. 7 after exposure to BCG for 23 hours. Then during the different culture time after discard the BCG in the supernatant, we analyzed the response of RAW264. 7 by flow cytometry using carboxyfluorescein diacetate, succinimidyl ester (CFDA-SE), annexin V/PI and Rhodamine 123, respectively.
  
  
  RESULTS
  We observed the phagosome containing BCG under transmission electronic microscope and RAW264. 7 cells still grow well after incubated with BCG for 23 hours. We found that the expression level of co-stimulatory molecules, CD40, CD54, CD80, CD86 and CD11b, were elevated evidently, except CD11c, I-A(d) and H-2K(d). The fluorescence intensity of BCG stained with CFDA-SE decreased with the prolongation of culture time, but it was still more higher than unstained control four days later. After removal of uninfected BCG, the percents of BCG-infected cell decreased and we didn't detected BCG-infected cells evidently for 60 h culture. Furthermore, we also found that there was no obvious cell apoptosis for all the time, and the mitochondrial membrane potential of BCG-infected cells increased early and then decreased to the same level as the uninfected control.
  
  
  CONCLUSIONS
  These results would be helpful to elucidate the mechanism of BCG vaccination.
authors.affiliation: Jiangsu Key Laboratory of Zoonesis, Yangzhou University, Yangzhou 225009, China.
authors.forename: Maozhi
authors.initials: M
authors.lastname: Hu
authors.forename: Yifang
authors.initials: Y
authors.lastname: Chen
authors.forename: Lu
authors.initials: L
authors.lastname: Han
authors.forename: Yan
authors.initials: Y
authors.lastname: Ji
authors.forename: Jiayu
authors.initials: J
authors.lastname: Zheng
authors.forename: Chuang
authors.initials: C
authors.lastname: Meng
authors.forename: Haixia
authors.initials: H
authors.lastname: Zhou
authors.forename: Xiang
authors.initials: X
authors.lastname: Chen
authors.forename: Xinan
authors.initials: X
authors.lastname: Jiao
authors.forename: Xiufan
authors.initials: X
authors.lastname: Liu
mesh_terms: D000818:Animals; D018122:B7-1 Antigen / Q000378:metabolism; D051940:B7-2 Antigen / Q000378:metabolism; D039481:CD11b Antigen / Q000378:metabolism; D019013:CD40 Antigens / Q000378:metabolism; D002460:Cell Line; D005434:Flow Cytometry; D005452:Fluoresceins; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism; D008264:Macrophages / Q000378:metabolism* / Q000382:microbiology* / Q000648:ultrastructure; D053078:Membrane Potential, Mitochondrial; D051379:Mice; D008807:Mice, Inbred BALB C; D008853:Microscopy; D046529:Microscopy, Electron, Transmission; D009163:Mycobacterium bovis / Q000254:growth & development* / Q000502:physiology / Q000648:ultrastructure; D010588:Phagosomes / Q000648:ultrastructure; D020112:Rhodamine 123; D013388:Succinimides
chemical_list: C087165:5-(6)-carboxyfluorescein diacetate succinimidyl ester; D018122:B7-1 Antigen; D051940:B7-2 Antigen; D039481:CD11b Antigen; D019013:CD40 Antigens; D005452:Fluoresceins; D013388:Succinimides; D018799:Intercellular Adhesion Molecule-1; D020112:Rhodamine 123