OMP31 of Brucella melitensis 16M impairs the apoptosis of macrophages triggered by TNF-α

Zhang,Ke; Wang,Hui; Guo,Fei; Yuan,Li; Zhang,Wanjiang; Wang,Yuanzhi; Chen,Chuangfu

doi:10.3892/etm.2016.3655

OMP31 of Brucella melitensis 16M impairs the apoptosis of macrophages triggered by TNF-α

Authors:
- Ke Zhang
- Hui Wang
- Fei Guo
- Li Yuan
- Wanjiang Zhang
- Yuanzhi Wang
- Chuangfu Chen
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Affiliations: Department of Basic Medicine, School of Medicine, Pingdingshan University, Pingdingshan, Henan 467000, P.R. China, Department of Geriatrics, Shihezi People's Hospital, Shihezi, Xinjiang 832000, P.R. China, Department of Basic Medicine, School of Medicine, Shihezi University, Shihezi, Xinjiang 832003, P.R. China, School of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang 832003, P.R. China
Published online on: September 2, 2016 https://doi.org/10.3892/etm.2016.3655
Pages: 2783-2789

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Abstract

Outer membrane proteins (OMPs) of microorganisms play important roles in directly interacting with host cells. Brucella species inhibit the apoptosis of host cells to benefit their own intracellular survival and replication. However, the association between OMP31 of Brucella and host cell apoptosis, and the underlying mechanism are unclear. In this study, an OMP31 gene deletion mutant based on B. melitensis 16M was constructed. Following the infection of RAW264.7 cells with B. melitensis 16M or the mutant strain, colony formation, apoptosis, tumor necrosis factor (TNF)‑α levels and the levels of key downstream factors of the apoptosis pathways triggered by TNF‑α, namely caspase‑3, ‑8 and ‑9, cytochrome c, B‑cell lymphoma 2 (Bcl‑2) and Bcl‑2‑associated X protein (Bax) were detected. The mutant strain was shown to have the same phenotype as the parent strain using traditional microbiological tests. However, the mutant strain had impaired intracellular survival, with higher levels of apoptosis and TNF‑α expression in infected RAW164.7 macrophages than the parent strain. The downstream factors of apoptosis triggered by TNF‑α, including increased caspase‑8, ‑3 and ‑9, cytochrome c and Bax, and decreased Bcl‑2, indicated that the classical and mitochondrial cell death pathways were involved. It may be concluded that OMP31 from Brucella inhibited apoptosis and benefitted the intracellular survival of this microorganism. Furthermore, TNF‑α may have served as a switch triggering classical death and mitochondrial cell death pathways.

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