Transcriptome Landscape of Intracellular Brucella ovis Surviving in RAW264.7 Macrophage Immune System
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ORIGINAL ARTICLE
Transcriptome Landscape of Intracellular Brucella ovis Surviving in RAW264.7 Macrophage Immune System Hanwei Jiao,1,2,3,4 Bowen Li,1,2,3 Zonglin Zheng,2 Zhixiong Zhou,1,2,3 Wenjie Li,1,2,3 Guojing Gu ,2 Juan Liu,1,2,3 Yichen Luo,1,2,3 Xuehong Shuai,1,2,3 Yu Zhao,1,2,3 Yuxuan Liu,2 Yidan Wang,2 Xinglong Wang,2 Xiaoyan Hu,2 Li Wu,2,3 Jixuan Chen,2,3 and Qingzhou Huang2,3
Abstract— Brucella ovis infection results in genital damage and epididymitis in rams, placental inflammation and rare abortion in ewes, and neonatal mortality in lambs. However, the mechanism underlying B. ovis infection remains unclear. In the present study, we used prokaryotic transcriptome sequencing to identify the differentially expressed genes (DEGs) between wild-type B. ovis and intracellular B. ovis in RAW264.7 macrophages. Gene ontology (GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed, and quantitative reverse transcriptase PCR (qRT-PCR) was used to validate the top 10 upregulated and downregulated DEGs. The results showed that 212 genes were differentially expressed, including 68 upregulated and 144 downregulated genes, which were mainly enriched in 30 GO terms linked to biological process, cellular component, and molecular function. KEGG analysis showed that the DEGs were enriched in the hypoxia-inducible factor 1 (HIF-1) signaling pathway, mitogen-activated protein kinase (MAPK) signaling pathway, beta-alanine metabolism, and quorum sensing pathway. BME_RS01160, BME_RS04270, BME_RS08185, BME_RS12880, BME_RS25875, predicted_RNA865, and predicted_RNA953 were confirmed with the transcriptome sequencing data. Hence, our findings not only reveal the intracellular parasitism of B. ovis in the macrophage immune system, but also help to understand the mechanism of chronic B. ovis infection. KEY WORDS: Brucella ovis; macrophage; immune system; intracellular; prokaryotic transcriptome sequencing.
INTRODUCTION Hanwei Jiao, Bowen Li and Zonglin Zheng contributed equally to this work. 1
Immunology Research Center, Medical Research Institute, Southwest University, Chongqing, 402460, People’s Republic of China 2 College of Animal Science, Southwest University, Chongqing, 402460, People’s Republic of China 3 Chongqing Veterinary Scientific Engineering Research Center, Southwest University, Chongqing, 402460, People’s Republic of China 4 To whom correspondence should be addressed at Immunology Research Center, Medical Research Institute, Southwest University, Chongqing, 402460, People’s Republic of China. E-mail: [email protected]
Brucella is a type of gram-negative, non-motile aerobic bacteria without a capsule (smooth type with microcapsule). Further, it is oxidase-positive, reduces nitrates, functions as an intracellular parasite, and can survive in various domestic animals [1–3]. Brucellosis, caused by Brucella, is a chronic infectious disease in both humans and animals. The World Health Organization (WHO) divided Brucella into the following six species according to their
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