| Autor: |
Xin PAN, Guang-bo LI, Han LI, Jia-lin CAI, Long CHEN, Xia-xian SHEN, Pei-pei LIU, Jian-jin WU |
| Jazyk: |
čínština |
| Rok vydání: |
2011 |
| Předmět: |
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| Zdroj: |
Medical Journal of Chinese People's Liberation Army, Vol 36, Iss 10, Pp 1023-1026 (2011) |
| ISSN: |
0577-7402 |
| Popis: |
Objective To investigation on the early carrying inducible nitric oxide synthase for peroxisomes to Salmonella typhimurium during the bacteria infection mouse macrophages.Methods RAW264.7 macrophages were transfected with pTassC-GFP plasmids to analysis the existence form of green fluorescent protein labeled target for Salmonella secreted protein SpiC(TassC)protein in the cell.The interaction between the fusion protein TassC-GFP and peroxisomes were analyzed by co-transfection of pTassC-GFP and pDsRed2-Perxi(labels peroxisomes red) plasmids to RAW264.7 macrophages,the positive transfected cells named RAW-DT.RAW-D cells were named by transfecting RAW264.7 with pDsRed2-Perxi plasmids.S.typhimurium was detected with mono-antibody and visualized with Alexa Fluor 350 conjugated donkey anti-mouse antibodies.Inducible nitric oxide synthase(iNOS or NOS2) was detected with iNOS-antibody and visualized with Alexa Fluor 488 conjugated goat anti-rabbit antibodies.S.typhimurium were used to infect the RAW-DT cells to analyze the interaction among bacteria,TassC-GFPs and peroxisomes.The RAW-D cells were infected with S.typhimurium 1h to analyze the interaction among bacteria,iNOS and peroxisomes.Results TassC vesicles co-localized with peroxisomes when RAW264.7 macrophages were co-transfected with pTassC-GFP and pDsRed2-Perxi plasmids.It was determined by a three-dimensional(xyz) fluorescence microscopy that the recruitment or overlapping of TassC-GFP and pemxiomes to the Salmonella-containing vacuoles(SCV) after infection of RAW-DT macrophages with S.typhimurium for 1h.The SCVs also could co-localized with peroxisomes and iNOS after infection of RAW-D cells with S.typhimurium for 1h.Upon entry of Salmonella,peroxisomes were recruited to the Salmonella-containing vesicles and remain aggregated around the SCV for the duration of the 60 minutes observation time.Conclusion These findings indicated that,wild type S.typhimurium could induce iNOS production in RAW264.7 macrophages,both of iNOS and TassC could bind with peroxisomes.Host cell TassC has been identified as the host cell target for S.typhimurium secreted SpiC.The findings suggest that if SpiC was still attached with bacteria,TassC would mediated peroxisomes binding with iNOS involved in the early germicidal phase of wild type S.typhimurium infection.If SpiC was secreted to the cytosol,the bactericidal effection of iNOS would be decrease. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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