BCG感染牛肺泡巨噬细胞的转录组测序和分析

doi:10.13481/j.1671-587x.20190103

吉林大学学报(医学版) ›› 2019, Vol. 45 ›› Issue (01): 12-16.doi: 10.13481/j.1671-587x.20190103

• 基础研究 • 上一篇

BCG感染牛肺泡巨噬细胞的转录组测序和分析

徐金瑞, 吕翠萍, 杨易, 周彦兵, 骆佳, 王玉炯

宁夏大学西部特色生物资源保护与利用教育部重点实验室, 宁夏银川 750021

收稿日期:2018-05-24 发布日期:2019-01-28
通讯作者: 王玉炯,教授,博士研究生导师(Tel:0951-2062033,E-mail:wyj@nxu.edu.cn) E-mail:wyj@nxu.edu.cn
作者简介:徐金瑞(1975-),男,宁夏回族自治区永宁县人,教授,遗传学硕士,主要从事病原微生物学方面的研究。

Transcriptomesequencing and analysis of bovine alveolar macrophages infected with BCG

XU Jinrui, LYU Cuiping, YANG Yi, ZHOU Yanbing, LUO Jia, WANG Yujiong

Key Laboratory of Protection and Utilization of Special Biological Resources in Western China, Ministry of Education, Ningxia University, Yinchuan 750021, China

Received:2018-05-24 Published:2019-01-28
Contact: 国家自然科学基金资助课题（31572494，31760733）；宁夏回族自治区科技厅自然科学基金资助课题（2018AAC03019）；宁夏回族自治区重点研发计划项目资助课题（东西部合作，2017BN04） E-mail:wyj@nxu.edu.cn
Supported by:
This work was supported by Fundaçaõ de Amparo à Pesquisa do Estado de Saõ Paulo (FAPESP) (Grant 2009/52237-9 to WV, fellowship 2014/23951-3 to LMSM).

摘要/Abstract

摘要： 目的：通过卡介苗（BCG）感染牛肺泡巨噬细胞（BAM）的转录组测序及生物信息学分析，揭示差异表达的基因和长链非编码RNA （lncRNA），为巨噬细胞抗结核分枝杆菌感染的免疫调控机制研究提供理论依据。方法：采用肺脏灌洗离心法收集并培养BAM，分为感染组和未感染组，感染组经BCG感染后12h，利用转录组测序技术（RNA-Seq）检测感染组和未感染组BAM mRNA表达谱及lncRNA表达谱，并进行生物信息学相关分析。结果：与未感染组比较，感染组BAM差异表达的lncRNAs共有119个（P<0.05），差异表达的mRNA共有1 111个（P<0.05）。在差异表达基因的功能富集分析中，最显著富集项与免疫功能相关（GO：0006955，P<0.05），共有125个基因，其中有63个基因表达上调，62个基因表达下调，且白细胞介素6（IL-6）、白细胞介素17（IL-17）和白细胞介素23A （IL-23A）等促炎因子表达上调。lncRNA靶基因预测，差异表达的lncRNA参与转化生长因子β（TGF-β）信号通路及ABC转运体信号通路的调控（P<0.05）。结论：BCG感染BAM后，激发宿主细胞产生强烈的免疫反应，引起lncRNA和mRNA表达谱的改变。

关键词: 卡介苗, 牛肺泡巨噬细胞, 转录组测序, 长链非编码RNA

Abstract: Objective: To reveal the differentially expressed genes and long non-coding RNAs (lncRNAs) by sequencing the transcriptome of bovine alveolar macrophages (BAM) infected with Bacillus Calmette-Guérin (BCG) and analyzing their bioinformations, and to provide theoretical foundation for the research on immune regulation mechanism of anti-infection of Mycobacterium tuberculosis of macrophages.Methods: The BAM were collected by pulmonary lavage and centrifugation and cultured and divided into infected group and uninfected group.After infection for 12 h in infected group,the expression profiles of mRNA and lncRNA in infected group and uninfected group were detected by RNA-Seq, and bioinformatics analysis was carried out.Results: compared with uninfected group, there were 119 differentially expressed lncRNAs and 1111 differentially expressed mRNA in infected group (P<0.05). Gene Ontology functional enrichment analysis showed that the most significant enrichment was immune response (GO:0006955, P<0.05), including 125 genes, in which 63 were up-regulated and 62 were down-regulated, and the expressions of proinflammatory factors interleukin-1 (IL-6), interleukin-7 (IL-7), and interleukin-23A (IL-23A) were up-regulated. Cis target gene prediction and KEGG pathway analysis showed that the differentially expressed lncRNAs were involved in the regulation of transforming growth factor-β (TGF-β) signaling pathway and ATP-binding cassette transporter (ABC transporter) signaling pathway (P<0.05).Conclusion: The host cells are stimulated to produce a strong immune response after the BAM are infected by BCG and results in the changes of lncRNA and mRNA expression profiles.

Key words: Bacillus Calmette-Guérin, bovine alveolar macrophage, transcriptome sequencing, long non-coding RNA

中图分类号:

Q939.91

徐金瑞, 吕翠萍, 杨易, 周彦兵, 骆佳, 王玉炯. BCG感染牛肺泡巨噬细胞的转录组测序和分析[J]. 吉林大学学报(医学版), 2019, 45(01): 12-16.

XU Jinrui, LYU Cuiping, YANG Yi, ZHOU Yanbing, LUO Jia, WANG Yujiong. Transcriptomesequencing and analysis of bovine alveolar macrophages infected with BCG[J]. Journal of Jilin University(Medicine Edition), 2019, 45(01): 12-16.

参考文献

[1] AN M,RYU D R,WON PARK J, et al. ULK1 prevents cardiac dysfunction in obesity through autophagy-meditated regulation of lipid metabolism[J]. Cardiovas Res, 2017, 113(10):1137-1147.
[2] WATERS WR,PALMER MV. Mycobacterium bovis infection of cattle and white-tailed deer:Translational research of relevance to human tuberculosis[J]. Ilar J, 2015, 56(1):26-43.
[3] OUYANG J,HU J,CHEN JL. lncRNAs regulate the innate immune response to viral infection[J]. Wiley Interdiscip Rev RNA, 2016, 7(1):129-143.
[4] FU Y, XU X, XUE J, et al. Deregulated lncRNAs in B cells from patients with active tuberculosis[J]. PLoS ONE, 2017, 12(1):1-14.
[5] YI Z, LI J, GAO K, et al. Identifcation of differentially expressed long non-coding RNAs in CD4+ T cells response to latent tuberculosis infection[J]. J Infect, 2014, 69(6):558-568.
[6] YANG X, YANG J, WANG J, et al. Microarray analysis of long noncoding RNA and mRNA expression profiles in human macrophages infected with ycobacterium tuberculosis[J]. Sci Rep, 2016,14(6):38963.
[7] MASTERS S L,MIELKE L A,CORNISH A L, et al. Regulation of interleukin-1beta by interferon-gamma is species specific, limited by suppressor of cytokine signalling 1 and influences interleukin-17 production[J]. EMBO Rep, 2010, 11(8):640-646.
[8] MAGEE D A,CONLON K M,NALPAS N C, et al. Innate cytokine profiling of bovine alveolar macrophages reveals commonalities and divergence in the response to Mycobacterium bovis and Mycobacterium tuberculosis infection[J]. Tuberculosis, 2014, 94(4):441-450.
[9] HMAMA Z,PENA-DIZA S, JOSEPH S, et al. Immunoevasion and immunosuppression of the macrophage by Mycobacterium tuberculosis[J]. Immunol Rev, 2015, 264(1):220-232.
[10] MORACO A H,KORNFELD H. Cell death and autophagy in tuberculosis[J]. Semin Immunol, 2014, 26(6):497-511.
[11] VAN DER WEL N,DAVA D,HOUBEN D, et al. M. tuberculosis and M. leprae translocate from the phagolysosome to the cytosol in myeloid cells[J]. Cell, 2007, 129(7):1287-1298.
[12] BACH H, PAPAVINASASUNDARAM K G, WONG D, et al. Mycobacterium tuberculosis virulence is mediated by PtpA dephosphorylation of human vacuolar protein sorting 33B[J]. Cell Host Microbe, 2008, 3(5):316-322.
[13] PAWAR K,HANISCH C,PALMA VERA S E, et al. Down regulated lncRNA MEG3 eliminates mycobacteria in macrophages via autophagy[J]. Sci Rep, 2016,14(6):19416.
[14] VAN DER DEEN M,DE VRIES E G,TIMENS W, et al. ATP-binding cassette (ABC) transporters in normal and pathological lung[J]. Respir Res, 2005.DOI:10.1186/1465-9921-6-59.
[15] YIN K,LIAO D F,TANG C K.. ATP-binding membrane cassette transporter A1(ABCA1):A possible link between inflammation and reverse cholesterol transport[J]. Mol Med, 2010, 16(9/10):438-449.
[16] CASTRILLO A,JOSEPH S B,VAIDYA S A, et al. Crosstalk between LXR and Toll-like receptor signaling mediates bacterial and viral antagonism of cholesterol metabolism[J]. Mol Cell, 2003, 12(4):805-816.
[17] KHOVIDHUNKIT W,KIM M S,MEMON R A, et al. Effects of infection and inflammation on lipid and lipoprotein metabolism:mechanisms and consequences to the host[J]. Lipid Res, 2004, 45(7):1169-1196.
[18] AZZAM K M,FESSLER M B. Crosstalk between reverse cholesterol transport and innate immunity[J]. Trends Endocrin Met, 2012, 23(4):169-178.
[19] 刘萍,张丽,庞楠楠,等.血清IFN-γ、TGF-β1、IL-10检测对儿童潜伏性结核感染的诊断价值[J].中国病原生物学杂志,2011,6(9):650-652.
[20] WARSINSKE H C,PIENAAR E,LINDERMAN J J, et al. Deletion of TGF-beta1 increases bacterial clearance by cytotoxic T cells in a tuberculosis granuloma model[J]. Front Immunol, 2017, 8:1843.

BCG感染牛肺泡巨噬细胞的转录组测序和分析

Transcriptomesequencing and analysis of bovine alveolar macrophages infected with BCG

RichHTML

PDF (PC)

赞

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 6

Metrics

本文评价

推荐阅读 0

[1]	黄琦, 沙辉, 陈龙, 吕龙龙, 徐声鸣, 张泽天, 赵松, 靳凤艳, 牛丰. 多发性骨髓瘤和慢性淋巴细胞白血病患者血浆lncRNA MALAT1、LincRNA-p21和GAS5的表达水平及其诊断意义[J]. 吉林大学学报(医学版), 2018, 44(02): 339-345.
[2]	刘相良, 袁铭成, 纪伟, 徐鑫宇, 梅起腾, 李理, 陈晓, 李薇. 长链非编码RNA在肺癌发生发展中作用的研究进展[J]. 吉林大学学报(医学版), 2017, 43(02): 450-453.
[3]	王朝义, 王强, 郑曰勇, 李聪, 郭敦伟, 唐华. 卵泡抑素对肿瘤恶病质小鼠骨骼肌消耗的影响及其作用机制[J]. 吉林大学学报(医学版), 2016, 42(04): 653-658.
[4]	龚萍, 李云, 罗睿, 李红, 谭钰嫔, 龙毅. 皮内注射灭活卡介苗对哮喘大鼠CD4⁺CD25⁺Treg细胞数量和CTLA-4 mRNA表达的影响[J]. 吉林大学学报(医学版), 2015, 41(03): 517-521.
[5]	李贺, 王华, 吴秀丽, 卫红飞, 孙陆果, 张永胜, 于永利, 王丽颖, 万敏. 卡介苗热休克蛋白70对肿瘤细胞裂解物免疫原性的影响[J]. J4, 2011, 37(1): 68-71.
[6]	董恩军，张灵霞，吴雪琼，张翠英，朱琳，陆阳. PstS1-卡介苗重组疫苗的构建[J]. J4, 2006, 32(2): 297-300.