[1]郭苗苗,尹 鑫,姚秋成,等.稳定表达马属动物SAMHD1蛋白的U937细胞系的建立及其抗病毒活性初步研究[J].中国预防兽医学报,2017,(10):831-835.[doi:10.3969/j.issn.1008-0425.201701002]
 GUO Miao-miao,YIN Xin,YAO Qiu-cheng,et al.Establishment of U937 cell lines stably expressing Equus SAMHD1[J].Chinese journal of preventive veterinary medicine,2017,(10):831-835.[doi:10.3969/j.issn.1008-0425.201701002]
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稳定表达马属动物SAMHD1蛋白的U937细胞系的建立及其抗病毒活性初步研究()
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2017年10
页码:
831-835
栏目:
免疫学
出版日期:
2017-10-15

文章信息/Info

Title:
Establishment of U937 cell lines stably expressing Equus SAMHD1
文章编号:
1008-0589(2017)10-0831-05
作者:
 

郭苗苗尹 鑫姚秋成胡 哲王晓钧*

 (中国农业科学院哈尔滨兽医研究所 兽医生物技术国家重点实验室/马传染病和慢病毒病研究创新团队,黑龙江 哈尔滨 150069)
Author(s):
 

GUO Miao-miao YIN Xin YAO Qiu-cheng HU Zhe WANG Xiao-jun*

 (State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute,
Chinese Academy of Agricultural Sciences, Harbin 150069,China)
关键词:
SAMHD1U937细胞系HIV-1EIAV
Keywords:
 equine  donkey  SAMHD1  U937 cell lines  HIV-1  EIAV
分类号:
S852.65
DOI:
10.3969/j.issn.1008-0425.201701002
文献标志码:
A
摘要:
为研究马属动物SAMHD1的抗病毒活性,本研究构建了稳定表达马SAMHD1 (eqSAMHD1)、驴SAMHD1 (doSAMHD1)的U937细胞系。利用pLPCX慢病毒表达载体构建重组质粒pLPCX-doSAMHD1-HA和 pLPCX-eqSAMHD1-HA。将重组质粒和假病毒包装重组质粒(pCgp和pVSV-G)共转染于293T细胞中,制备表达SAMHD1的假病毒,将其感染人急性单核细胞白血病细胞系U937,利用嘌呤霉素筛选,建立了稳定表达doSAMHD1和eqSAMHD1的U937细胞系。经western blot鉴定,该细胞系能够稳定表达doSAMHD1和eqSAMHD1蛋白。将人类免疫缺陷病毒(HIV)和马传染性贫血病毒(EIAV)的假病毒感染该细胞系,结果显示表达eqSAMHD1和doSAMHD1的细胞系均能够显著抑制HIV假病毒复制,限制倍数分别为17~19倍和14~18倍;两种细胞系也能够抑制EIAV假病毒复制,限制倍数分别为3~4倍和2~3倍。该细胞系的建立为进一步评价和研究马属动物SAMHD1的抗病毒机制奠定了基础。
  
Abstract:
The SAM domain and HD domain-containing protein 1 (SAMHD1) is a host restriction that inhibits HIV-1 replication in dendritic cells, monocytes, and macrophages. We found equus also encode SAMHD1 proteins. To evaluate the antiviral activity of equine and donkey SAMHD1, two recombinant plasmids containing the equine SAMHD1 and donkey SAMHD1 genes based on pLPCX plasmid were constructed, respectively. The pseudoviruses were produced by co-transfected HEK293T cell with recombinant plasmids and lentivirus packaging recombinant plasmid (pCgp and pVSV-G). Then cell lines were established by infecting U937 cell with the pseudovirus, designed U937-eqSAMHD1 and U937-doSAMHD1, which were confirmed by western blot. To test their anti-retroviral activity, the cell lines were infected with the HIV-Luc or EIAV-Luc pseudovirus. The results showed that U937-eqSAMHD1 and U937-doSAMHD1 could  restrict human immunodeficiency virus (HIV) by 17 to 19-fold and 14 to 18-fold. While they could slightly restrict equine infectious anemia virus (EIAV) infection, by 3 to 4-fold and 2 to 3-fold. Our results provided evidence that eqSAMHD1 and doSAMHD1 could restrict HIV-1 and EIAV replication and help to further study on restriction mechanism of lentivirus infection.
 

参考文献/References:

[1] 杨光文,张金阳,夏雪山. 具有广泛抗病毒活性的SAMHD1蛋白的研究进展[J]. 生命科学研究,2014,18(4):367-371.
[2]Gramberg T, Kahle T, Bloch N, et al. Restriction of diverse retroviruses by SAMHD1 [J]. Retrovirology, 2013, 10: 26.
[3]Hrecka K, Hao Cai-li, Gierszewska M, et al. Vpx relieves inhibition of HIV-1 infection of macrophages mediated by the SAMHD1 protein [J]. Nature, 2011, 474(7353): 658-661.
[4]Goldstone D C, Ennis-Adeniran V, Hedden J J, et al. HIV-1 restriction factor SAMHD1 is a deoxynucleoside triphosphate triphosphohydrolase [J]. Nature, 2011, 480(7377): 379-382.
[5]Baldauf H M, Pan Xiao-yu, Erikson E, et al. SAMHD1 restricts HIV-1 infection in resting CD4(+) T cells [J]. Nat Med, 2012, 18(11): 1682-1687.
[6]Welbourn S and Strebel K. Low dNTP levels are necessary but may not be sufficient for lentiviral restriction by SAMHD1 [J]. Virology, 2016, 488: 271-277.
[7]Hollenbaugh J A, Gee P, Baker J, et al. Host factor SAMHD1 restricts DNA viruses in non-dividing myeloid cells [J]. PLoS Pathog, 2013, 9(6): e1003481.
[8]Kim E T, White T E, Brandariz-Nunez A, et al. SAMHD1 restricts herpes simplex virus 1 in macrophages by limiting DNA replication [J]. J Virol, 2013, 87(23): 12949-12956.
[9]Kim B, Nguyen L A, Daddacha W, et al. Tight interplay among SAMHD1 protein level, cellular dNTP levels, and HIV-1 proviral DNA synthesis kinetics in human primary monocyte-derived macrophages [J]. J Biol Chem, 2012, 287(26): 21570-21574.
[10]Lahouassa H, Daddacha W, Hofmann H, et al. SAMHD1 restricts HIV-1 by reducing the intracellular pool of deoxynucleotide triphosphates [J]. Nat Immunol, 2012, 13: 223-228.
[11]Beloglazova N, Flick R, Tchigvintsev A, et al. Nuclease activity of the human SAMHD1 protein implicated in the Aicardi-Goutieres syndrome and HIV-1 restriction [J]. J Biol Chem, 2013, 288(12): 8101-8110.
[12]Cribier A, Descours B, Valadao A L, et al. Phosphorylation of SAMHD1 by cyclin []A2/CDK1 regulates its restriction activity toward HIV-1 [J]. Cell Rep, 2013, 3(4): 1036-1043.
[13]Welbourn S, Dutta S M, Semmes O J, et al. Restriction of virus infection but not catalytic dNTPase activity is regulated by phosphorylation of SAMHD1 [J]. J Virol, 2013, 87(21): 11516- 11524.
[14]White T E, Brandariz-Nunez A, Valle-Casuso J C, et al. The retroviral restriction ability of SAMHD1, but not its deoxynucleotide triphosphohydrolase activity, is regulated by phosphorylation [J]. Cell Host Microbe, 2013, 13(4): 441-451.
[15]Berger G, Turpin J, Cordeil S, et al. Functional analysis of the relationship between Vpx and the restriction factor SAMHD1 [J]. J Biol Chem, 2012, 287(49): 41210-41217.
[16]Ahn J, Hao Cai-li, Yan Jun-peng, et al. HIV/simian immunodeficiency virus (SIV) accessory virulence factor Vpx loads the host cell restriction factor SAMHD1 onto the E3 ubiquitin ligase complex CRL4DCAF1 [J]. J Biol Chem, 2012, 287(15): 12550-12558.
[17]Rice G I, Bond J, Asipu A, et al. Mutations involved in Aicardi-Goutieres syndrome implicate SAMHD1 as regulator of the innate immune response [J]. Nat Genet, 2009, 41(7): 829-832.
[18]Ayinde D, Casartelli N, and Schwartz O. Restricting HIV the SAMHD1 way: through nucleotide starvation [J]. Nat Rev Microbiol, 2012, 10(10): 675-680.
[19]Zhu Chun-feng, Gao Wen-ying, Zhao Ke, et al. Structural insight into dGTP-dependent activation of tetrameric SAMHD1 deoxynucleoside triphosphate triphosphohydrolase [J]. Nat Commun, 2013, 4: 2722.
[20]Wittmann S, Behrendt R, Eissmann K, et al. Phosphorylation of murine SAMHD1 regulates its antiretroviral activity [J]. Retrovirology, 2015, 12: 103.
(本文编辑:李 娜)

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更新日期/Last Update: 2017-11-06