title: [Infection of recombinant adenovirus Ad5F35-IL-12 in different kinds of human mononuclear macrophages].
abstract: Objective To investigate the efficiencies of transfection and expression of human recombinant adenovirus Ad5F35-IL-12 in the different kinds of human  mononuclear macrophages.  Methods The human recombinant adenovirus Ad5F35-IL-12 was used to infect human peripheral blood monocytes,  pleural fluid macrophages as well as THP-1,  U937 monocyte cell lines and their phorbol myristate acetate (PMA)-induced macrophages. 48 h later,  green fluorescence was observed under the fluorescence microscope to detect the transfection efficiency. The expressions of IL-12 double-subunits (p35,  p40) mRNA were tested by RT-PCR and the level of IL-12p70 protein in the cell culture supernatant was detected with ELISA.  Results The human recombinant adenovirus Ad5F35-IL-12 successfully infected the human peripheral blood monocytes,  pleural fluid macrophages,  THP-1 monocytes,  U937 monocytes,  and THP-1 and U937 macrophages induced with PMA. All above infected mononuclear macrophages effectively secreted IL-12p70 protein,  and they were listed from high to low of IL-12p70 protein level as pleural fluid macrophages,  U937 and THP-1 macrophages induced with PMA,  U937 monocytes,  human peripheral blood monocytes and THP-1 monocytes.  Conclusion The human recombinant adenovirus Ad5F35-IL-12 could infect different kinds of mononuclear macrophages,  and IL-12 p70 protein could be successfully expressed in cell supernatants.
authors: Chongqing Medical University, Chongqing, China.
authors: Qin
authors: Q
authors: Wang
authors: Feng
authors: F
authors: Cheng
authors: Weili
authors: W
authors: Zhang
authors: Wenbin
authors: W
authors: Kuang
authors: Pu
authors: P
authors: Li
authors: Jinyu
authors: J
authors: Dong
authors: Qindong
authors: Q
authors: Liang
authors: Zhiguang
authors: Z
authors: Tu
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D001117:Arenaviridae Infections / Q000235:genetics*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D018664:Interleukin-12 / Q000096:biosynthesis / Q000276:immunology; D008264:Macrophages / Q000276:immunology / Q000821:virology*; D011994:Recombinant Proteins / Q000235:genetics
chemical_list: D011994:Recombinant Proteins; D018664:Interleukin-12