Shi-jun Li1, Ye Hu, Jie Yan, Ya-fei Mao, Li-wei Li. 1. Department of Microbiology and Parasitology, College of Medicine, Zhejiang University, Hangzhou 310058, China.
Abstract
OBJECTIVE: To determine the involvement of FasL/Fas pathway in apoptosis of J774A.1 cells induced by Leptospira interrogans. METHODS: The cell infection model was established with mouse monocyte-macrophage J774A.1 cells infected by L.interrogans serogroup Icterohaemorrhagiae serovar lai strain 56601. The morphological characteristics of apoptotic J774A.1 cells were observed by DAPI staining method, and the apoptosis rate was quantitatively determined by flow cytometry. FasL neutralizing antibody was applied to block the apoptosis. Expression of FasL or Fas in the L.interrogans strain 56601-infected J774A.1 cells was detected by flow cytometry using PE-conjugated monoclonal antibody. RESULT: Chromatin condensation and marginalization were found in J774A.1 cells infected by L.interrogans strain 56601 for 4 h, which became more predominant for 24 h and karyorrhexis was present in some cells. When J774A.1 cells were infected for 4 h and 24 h, the apoptosis rates were 53.6% and 64.31%, respectively. However, the apoptosis rates were decreased to 10.27% and 15.9% after the cells were pre-treated with FasL neutralizing antibody. When J774A.1 cells were infected for 4 h and 24 h, FasL expression rates were increased to 21.69% and 65.70% from that of 4.19% before infection, and Fas expression rates were risen to 91.96% and 88.01% from that of 12.88% before infection. CONCLUSION: Inducement of cell apoptosis is an important mechanism of L.interrogans strain 56601 injuring J774A.1 cells. The strain of L.interrogans is able to up-regulate FasL/Fas expression levels of host cells and induce apoptosis of the cells via FasL/Fas pathway.
OBJECTIVE: To determine the involvement of FasL/Fas pathway in apoptosis of J774A.1 cells induced by Leptospira interrogans. METHODS: The cell infection model was established with mouse monocyte-macrophage J774A.1 cells infected by L.interrogans serogroup Icterohaemorrhagiae serovar lai strain 56601. The morphological characteristics of apoptotic J774A.1 cells were observed by DAPI staining method, and the apoptosis rate was quantitatively determined by flow cytometry. FasL neutralizing antibody was applied to block the apoptosis. Expression of FasL or Fas in the L.interrogans strain 56601-infected J774A.1 cells was detected by flow cytometry using PE-conjugated monoclonal antibody. RESULT: Chromatin condensation and marginalization were found in J774A.1 cells infected by L.interrogans strain 56601 for 4 h, which became more predominant for 24 h and karyorrhexis was present in some cells. When J774A.1 cells were infected for 4 h and 24 h, the apoptosis rates were 53.6% and 64.31%, respectively. However, the apoptosis rates were decreased to 10.27% and 15.9% after the cells were pre-treated with FasL neutralizing antibody. When J774A.1 cells were infected for 4 h and 24 h, FasL expression rates were increased to 21.69% and 65.70% from that of 4.19% before infection, and Fas expression rates were risen to 91.96% and 88.01% from that of 12.88% before infection. CONCLUSION: Inducement of cell apoptosis is an important mechanism of L.interrogans strain 56601 injuring J774A.1 cells. The strain of L.interrogans is able to up-regulate FasL/Fas expression levels of host cells and induce apoptosis of the cells via FasL/Fas pathway.