- Lon Mutant of Brucella abortus Induces Tumor Necrosis Factor-Alpha in Murine J774.A1 Macrophage
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Sungdo Park, Young-Sill Choi, Sang-Hee Park, Young-Rok Kim, Hyuk Chu, Kyu-Jam Hwang, Mi-Yeoun Park
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Osong Public Health Res Perspect. 2013;4(6):301-307. Published online December 31, 2013
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DOI: https://doi.org/10.1016/j.phrp.2013.10.002
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 Abstract
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- Objectives
The objective of this study was to isolate a Brucella lon mutant and to analyze the cytokine response of B. lon mutant during macrophage infection.
Methods
A wild-type Brucella abortus strain was mutagenized by Tn5 transposition. From the mouse macrophage J774.A1 cells, total RNA was isolated at 0 hours, 6 hours, 12 hours, and 24 hours after infection with Brucella. Using mouse cytokine microarrays, we measured transcriptional levels of the cytokine response, and validated our results with a reverse transcriptase-polymerase chain reaction (RT-PCR) assay to confirm the induction of cytokine messenger RNA (mRNA).
Results
In host J774.A1 macrophages, mRNA levels of T helper 1 (Th1)-type cytokines, including tumor necrosis factor-alpha (TNF-α), interferon-gamma (IFN-γ), interleukin-2 (IL-2), and IL-3, were significantly higher in the lon mutant compared to wild-type Brucella and the negative control. TNF-α levels in cell culture media were induced as high as 2 μg/mL after infection with the lon mutant, a greater than sixfold change.
Conclusion
In order to understand the role of the lon protein in virulence, we identified and characterized a novel B. lon mutant. We compared the immune response it generates to the wild-type Brucella response in a mouse macrophage cell line. We demonstrated that the B. lon mutants induce TNF-α expression from the host J774.A1 macrophage.
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Citations
Citations to this article as recorded by  - Structure, Substrate Specificity and Role of Lon Protease in Bacterial Pathogenesis and Survival
Perumalraja Kirthika, Khristine Kaith Sison Lloren, Vijayakumar Jawalagatti, John Hwa Lee
International Journal of Molecular Sciences.2023; 24(4): 3422. CrossRef - ClpP protease modulates bacterial growth, stress response, and bacterial virulence in Brucella abortus
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Veterinary Research.2023;[Epub] CrossRef - Brucella abortus Encodes an Active Rhomboid Protease: Proteome Response after Rhomboid Gene Deletion
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Proteomes.2020; 8(2): 8. CrossRef - Outer Membrane Vesicles From Brucella melitensis Modulate Immune Response and Induce Cytoskeleton Rearrangement in Peripheral Blood Mononuclear Cells
Eric Daniel Avila-Calderón, Olín Medina-Chávez, Leopoldo Flores-Romo, José Manuel Hernández-Hernández, Luis Donis-Maturano, Ahidé López-Merino, Beatriz Arellano-Reynoso, Ma. Guadalupe Aguilera-Arreola, Enrico A. Ruiz, Zulema Gomez-Lunar, Sharon Witonsky,
Frontiers in Microbiology.2020;[Epub] CrossRef - RNA-seq reveals the critical role of Lon protease in stress response and Brucella virulence
Yufu Liu, Hao Dong, Xiaowei Peng, Qiang Gao, Hui Jiang, Guanlong Xu, Yuming Qin, Jianrui Niu, Shijing Sun, Peng Li, Jiabo Ding, Ruiai Chen
Microbial Pathogenesis.2019; 130: 112. CrossRef - Brucella Downregulates Tumor Necrosis Factor-α to Promote Intracellular Survival via Omp25 Regulation of Different MicroRNAs in Porcine and Murine Macrophages
Xiaomao Luo, Xiujuan Zhang, Xingchen Wu, Xuefeng Yang, Cong Han, Zhengyu Wang, Qian Du, Xiaomin Zhao, Shan-Lu Liu, Dewen Tong, Yong Huang
Frontiers in Immunology.2018;[Epub] CrossRef
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