Aedes aegypti (Aag2)-derived clonal mosquito cell lines reveal the effects of pre-existing persistent infection with the insect-specific bunyavirus Phasi Charoen-like virus on arbovirus replication.
Aedes
/ virology
Alphavirus
/ growth & development
Animals
Arboviruses
/ genetics
Base Sequence
Cell Culture Techniques
/ methods
Cell Line
Coinfection
/ virology
Dengue Virus
/ growth & development
Flavivirus
/ genetics
Genome, Viral
Host-Pathogen Interactions
/ physiology
Mosquito Vectors
/ virology
Orthobunyavirus
/ genetics
RNA Viruses
/ genetics
Rhabdoviridae
/ growth & development
Sindbis Virus
/ growth & development
Transfection
Virus Replication
Zika Virus
/ growth & development
Journal
PLoS neglected tropical diseases
ISSN: 1935-2735
Titre abrégé: PLoS Negl Trop Dis
Pays: United States
ID NLM: 101291488
Informations de publication
Date de publication:
11 2019
11 2019
Historique:
received:
22
03
2019
accepted:
24
10
2019
revised:
18
11
2019
pubmed:
7
11
2019
medline:
19
2
2020
entrez:
7
11
2019
Statut:
epublish
Résumé
Aedes aegypti is a vector mosquito of major public health importance, transmitting arthropod-borne viruses (arboviruses) such as chikungunya, dengue, yellow fever and Zika viruses. Wild mosquito populations are persistently infected at high prevalence with insect-specific viruses that do not replicate in vertebrate hosts. In experimental settings, acute infections with insect-specific viruses have been shown to modulate arbovirus infection and transmission in Ae. aegypti and other vector mosquitoes. However, the impact of persistent insect-specific virus infections, which arboviruses encounter more commonly in nature, has not been investigated extensively. Cell lines are useful models for studying virus-host interactions, however the available Ae. aegypti cell lines are poorly defined and heterogenous cultures. We generated single cell-derived clonal cell lines from the commonly used Ae. aegypti cell line Aag2. Two of the fourteen Aag2-derived clonal cell lines generated harboured markedly and consistently reduced levels of the insect-specific bunyavirus Phasi Charoen-like virus (PCLV) known to persistently infect Aag2 cells. In contrast to studies with acute insect-specific virus infections in cell culture and in vivo, we found that pre-existing persistent PCLV infection had no major impact on the replication of the flaviviruses dengue virus and Zika virus, the alphavirus Sindbis virus, or the rhabdovirus vesicular stomatitis virus. We also performed a detailed characterisation of the morphology, transfection efficiency and immune status of our Aag2-derived clonal cell lines, and have made a clone that we term Aag2-AF5 available to the research community as a well-defined cell culture model for arbovirus-vector interaction studies. Our findings highlight the need for further in vivo studies that more closely recapitulate natural arbovirus transmission settings in which arboviruses encounter mosquitoes harbouring persistent rather than acute insect-specific virus infections. Furthermore, we provide the well-characterised Aag2-derived clonal cell line as a valuable resource to the arbovirus research community.
Sections du résumé
BACKGROUND
Aedes aegypti is a vector mosquito of major public health importance, transmitting arthropod-borne viruses (arboviruses) such as chikungunya, dengue, yellow fever and Zika viruses. Wild mosquito populations are persistently infected at high prevalence with insect-specific viruses that do not replicate in vertebrate hosts. In experimental settings, acute infections with insect-specific viruses have been shown to modulate arbovirus infection and transmission in Ae. aegypti and other vector mosquitoes. However, the impact of persistent insect-specific virus infections, which arboviruses encounter more commonly in nature, has not been investigated extensively. Cell lines are useful models for studying virus-host interactions, however the available Ae. aegypti cell lines are poorly defined and heterogenous cultures.
METHODOLOGY/PRINCIPLE FINDINGS
We generated single cell-derived clonal cell lines from the commonly used Ae. aegypti cell line Aag2. Two of the fourteen Aag2-derived clonal cell lines generated harboured markedly and consistently reduced levels of the insect-specific bunyavirus Phasi Charoen-like virus (PCLV) known to persistently infect Aag2 cells. In contrast to studies with acute insect-specific virus infections in cell culture and in vivo, we found that pre-existing persistent PCLV infection had no major impact on the replication of the flaviviruses dengue virus and Zika virus, the alphavirus Sindbis virus, or the rhabdovirus vesicular stomatitis virus. We also performed a detailed characterisation of the morphology, transfection efficiency and immune status of our Aag2-derived clonal cell lines, and have made a clone that we term Aag2-AF5 available to the research community as a well-defined cell culture model for arbovirus-vector interaction studies.
CONCLUSIONS/SIGNIFICANCE
Our findings highlight the need for further in vivo studies that more closely recapitulate natural arbovirus transmission settings in which arboviruses encounter mosquitoes harbouring persistent rather than acute insect-specific virus infections. Furthermore, we provide the well-characterised Aag2-derived clonal cell line as a valuable resource to the arbovirus research community.
Identifiants
pubmed: 31693659
doi: 10.1371/journal.pntd.0007346
pii: PNTD-D-19-00489
pmc: PMC6860454
doi:
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
e0007346Subventions
Organisme : Medical Research Council
ID : MR/R010315/1
Pays : United Kingdom
Organisme : Wellcome Trust
ID : 096062
Pays : United Kingdom
Organisme : NIAID NIH HHS
ID : U19 AI118610
Pays : United States
Organisme : NIAID NIH HHS
ID : R01 AI073450
Pays : United States
Organisme : Wellcome Trust
Pays : United Kingdom
Organisme : NIAID NIH HHS
ID : R21 AI116022
Pays : United States
Organisme : Medical Research Council
ID : G0801973
Pays : United Kingdom
Organisme : Wellcome Trust
ID : 096062
Pays : United Kingdom
Déclaration de conflit d'intérêts
The authors have declared that no competing interests exist.
Références
J Virol. 2009 May;83(9):4462-8
pubmed: 19224998
Nat Rev Microbiol. 2004 Oct;2(10):789-801
pubmed: 15378043
Virology. 2018 Jun 26;521:175-180
pubmed: 29957338
J Virol. 2003 Jan;77(2):1501-11
pubmed: 12502864
mSphere. 2017 May 3;2(3):
pubmed: 28497119
PLoS Negl Trop Dis. 2010 Oct 26;4(10):e856
pubmed: 21049065
Parasit Vectors. 2016 Jul 25;9(1):414
pubmed: 27457250
PLoS Negl Trop Dis. 2017 Oct 17;11(10):e0006010
pubmed: 29040304
Trans R Soc Trop Med Hyg. 1952 Sep;46(5):509-20
pubmed: 12995440
Annu Rev Med. 2018 Jan 29;69:395-408
pubmed: 28846489
Emerg Microbes Infect. 2018 Nov 15;7(1):181
pubmed: 30429457
Nucleic Acids Res. 2015 Jan;43(Database issue):D707-13
pubmed: 25510499
J Insect Physiol. 2013 Feb;59(2):159-70
pubmed: 22824741
BMC Genomics. 2017 Jul 5;18(1):512
pubmed: 28676109
Virology. 1975 Apr;64(2):367-77
pubmed: 806166
J Virol. 2018 Aug 16;92(17):
pubmed: 29950416
PLoS Negl Trop Dis. 2013 Jul 04;7(7):e2295
pubmed: 23861987
Nat Immunol. 2013 Apr;14(4):396-403
pubmed: 23435119
Virol J. 2014 May 20;11:97
pubmed: 24884700
Infect Genet Evol. 2019 Jan;67:191-209
pubmed: 30465912
Methods Enzymol. 2000;322:492-508
pubmed: 10914042
PLoS Negl Trop Dis. 2017 Jan 12;11(1):e0005187
pubmed: 28081143
Clin Microbiol Rev. 2005 Oct;18(4):608-37
pubmed: 16223950
J Gen Virol. 2012 Feb;93(Pt 2):223-234
pubmed: 22012464
Insect Mol Biol. 2010 Aug;19(4):441-9
pubmed: 20456509
Proc Natl Acad Sci U S A. 2012 Jan 3;109(1):255-60
pubmed: 22123944
Parasit Vectors. 2014 Aug 20;7:382
pubmed: 25141888
BMC Genomics. 2017 Jan 19;18(1):101
pubmed: 28103802
Virology. 2018 May;518:406-413
pubmed: 29625404
Parasit Vectors. 2018 May 23;11(1):310
pubmed: 29792223
Virology. 2012 Jun 5;427(2):90-7
pubmed: 22425062
J Virol. 2019 Aug 28;93(18):
pubmed: 31243123
PLoS One. 2013;8(2):e56534
pubmed: 23460804
Virology. 2015 Oct;484:51-8
pubmed: 26068885
Proc Natl Acad Sci U S A. 2012 Sep 4;109(36):14622-7
pubmed: 22908261
J Virol. 2017 Jul 12;91(15):
pubmed: 28539440
Nat Methods. 2012 Jun 28;9(7):676-82
pubmed: 22743772
Cancer Cell. 2003 Oct;4(4):263-75
pubmed: 14585354
Proc Biol Sci. 2013 Jan 22;280(1751):20122437
pubmed: 23193131
J Med Entomol. 2015 Mar;52(2):222-9
pubmed: 26336307
Parasit Vectors. 2012 Jul 24;5:148
pubmed: 22827926
Curr Biol. 2017 Nov 20;27(22):3511-3519.e7
pubmed: 29129531
Nature. 2018 Jul;559(7715):490-497
pubmed: 30046071
J Gen Virol. 2009 Nov;90(Pt 11):2669-2678
pubmed: 19656970
Virus Genes. 2019 Apr;55(2):127-137
pubmed: 30632016
Cell. 2009 Dec 24;139(7):1268-78
pubmed: 20064373
Vector Borne Zoonotic Dis. 2011 Aug;11(8):1099-105
pubmed: 21254845
Microbiol Mol Biol Rev. 2000 Mar;64(1):115-37
pubmed: 10704476
Sci Rep. 2017 Jul 31;7(1):6935
pubmed: 28761113
Proc Natl Acad Sci U S A. 2009 Oct 20;106(42):17841-6
pubmed: 19805194
PLoS Pathog. 2010 Apr 01;6(4):e1000833
pubmed: 20368968
PLoS Pathog. 2008 Jul 04;4(7):e1000098
pubmed: 18604274
Proc Natl Acad Sci U S A. 1996 May 14;93(10):4810-5
pubmed: 8643485
J Gen Virol. 2014 Dec;95(Pt 12):2796-2808
pubmed: 25146007
J Gen Virol. 2012 Jun;93(Pt 6):1215-1225
pubmed: 22377581
J Gen Virol. 2018 Dec;99(12):1739-1745
pubmed: 30394867
J Virol. 2009 Sep;83(18):9532-40
pubmed: 19570865
Nat Commun. 2016 Sep 01;7:12410
pubmed: 27580708
Southeast Asian J Trop Med Public Health. 2013 Jul 4;44(4):586-93
pubmed: 24050091
Viruses. 2019 Mar 18;11(3):
pubmed: 30889941
Curr Opin Virol. 2016 Dec;21:81-86
pubmed: 27580489
Nature. 2011 Aug 24;476(7361):450-3
pubmed: 21866159
Am J Trop Med Hyg. 1990 Dec;43(6):669-76
pubmed: 2267971
PLoS Pathog. 2012 Jan;8(1):e1002470
pubmed: 22241995
Heliyon. 2018 Aug 23;4(8):e00736
pubmed: 30167494
Virology. 2018 Oct;523:35-40
pubmed: 30077072
J Gen Virol. 2004 Jul;85(Pt 7):1971-1980
pubmed: 15218182
Viruses. 2015 Sep 10;7(9):4911-28
pubmed: 26378568
Biochem Pharmacol. 2017 Aug 15;138:7-18
pubmed: 28396196
Genes Dev. 2015 Oct 15;29(20):2168-82
pubmed: 26450910
Yale J Biol Med. 1970 Apr;42(5):276-92
pubmed: 5464392
Viruses. 2015 Nov 11;7(11):5801-12
pubmed: 26569286
Sci Rep. 2018 Mar 16;8(1):4690
pubmed: 29549363
J Virol. 2019 Mar 5;93(6):
pubmed: 30567990
J Gen Virol. 2006 Apr;87(Pt 4):735-48
pubmed: 16528021
Front Cell Infect Microbiol. 2017 Feb 03;7:22
pubmed: 28217557
Arch Virol. 2009;154(1):153-8
pubmed: 19089585
Virology. 2004 Nov 24;329(2):261-9
pubmed: 15518806
Nucleic Acids Res. 2014 Jul;42(13):8732-44
pubmed: 24939903
PLoS Negl Trop Dis. 2010 May 04;4(5):e671
pubmed: 20454569
PLoS Negl Trop Dis. 2010 Oct 26;4(10):e848
pubmed: 21049014