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Whole-Body Analysis of a Viral Infection: Vascular Endothelium is a Primary Target of Infectious Hematopoietic Necrosis Virus in Zebrafish Larvae


The progression of viral infections is notoriously difficult to follow in whole organisms. The small, transparent zebrafish larva constitutes a valuable system to study how pathogens spread. We describe here the course of infection of zebrafish early larvae with a heat-adapted variant of the Infectious Hematopoietic Necrosis Virus (IHNV), a rhabdovirus that represents an important threat to the salmonid culture industry. When incubated at 24°C, a permissive temperature for virus replication, larvae infected by intravenous injection died within three to four days. Macroscopic signs of infection followed a highly predictable course, with a slowdown then arrest of blood flow despite continuing heartbeat, followed by a loss of reactivity to touch and ultimately by death. Using whole-mount in situ hybridization, patterns of infection were imaged in whole larvae. The first infected cells were detectable as early as 6 hours post infection, and a steady increase in infected cell number and staining intensity occurred with time. Venous endothelium appeared as a primary target of infection, as could be confirmed in fli1:GFP transgenic larvae by live imaging and immunohistochemistry. Disruption of the first vessels took place before arrest of blood circulation, and hemorrhages could be observed in various places. Our data suggest that infection spread from the damaged vessels to underlying tissue. By shifting infected fish to a temperature of 28°C that is non-permissive for viral propagation, it was possible to establish when virus-generated damage became irreversible. This stage was reached many hours before any detectable induction of the host response. Zebrafish larvae infected with IHNV constitute a vertebrate model of an hemorrhagic viral disease. This tractable system will allow the in vivo dissection of host-virus interactions at the whole organism scale, a feature unrivalled by other vertebrate models.


Vyšlo v časopise: Whole-Body Analysis of a Viral Infection: Vascular Endothelium is a Primary Target of Infectious Hematopoietic Necrosis Virus in Zebrafish Larvae. PLoS Pathog 7(2): e32767. doi:10.1371/journal.ppat.1001269
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1001269

Souhrn

The progression of viral infections is notoriously difficult to follow in whole organisms. The small, transparent zebrafish larva constitutes a valuable system to study how pathogens spread. We describe here the course of infection of zebrafish early larvae with a heat-adapted variant of the Infectious Hematopoietic Necrosis Virus (IHNV), a rhabdovirus that represents an important threat to the salmonid culture industry. When incubated at 24°C, a permissive temperature for virus replication, larvae infected by intravenous injection died within three to four days. Macroscopic signs of infection followed a highly predictable course, with a slowdown then arrest of blood flow despite continuing heartbeat, followed by a loss of reactivity to touch and ultimately by death. Using whole-mount in situ hybridization, patterns of infection were imaged in whole larvae. The first infected cells were detectable as early as 6 hours post infection, and a steady increase in infected cell number and staining intensity occurred with time. Venous endothelium appeared as a primary target of infection, as could be confirmed in fli1:GFP transgenic larvae by live imaging and immunohistochemistry. Disruption of the first vessels took place before arrest of blood circulation, and hemorrhages could be observed in various places. Our data suggest that infection spread from the damaged vessels to underlying tissue. By shifting infected fish to a temperature of 28°C that is non-permissive for viral propagation, it was possible to establish when virus-generated damage became irreversible. This stage was reached many hours before any detectable induction of the host response. Zebrafish larvae infected with IHNV constitute a vertebrate model of an hemorrhagic viral disease. This tractable system will allow the in vivo dissection of host-virus interactions at the whole organism scale, a feature unrivalled by other vertebrate models.


Zdroje

1. KantherM

RawlsJF

2010 Host-microbe interactions in the developing zebrafish. Curr Opin Immunol 22 10 19

2. LamS

ChuaH

GongZ

LamT

SinY

2003 Development and maturation of the immune system in zebrafish, Danio rerio: a gene expression profiling, in situ hybridization and immunological study. Dev Comp Immunol 28 9 28

3. HerbomelP

ThisseB

ThisseC

1999 Ontogeny and behaviour of early macrophages in the zebrafish embryo. Development 126 3735 3745

4. Le GuyaderD

ReddMJ

Colucci-GuyonE

MurayamaE

KissaK

2008 Origins and unconventional behavior of neutrophils in developing zebrafish. Blood 111 132 141

5. RobertsenB

2006 The interferon system of teleost fish. Fish Shellfish Immunol 20 172 191

6. LevraudJ-P

BoudinotP

ColinI

BenmansourA

PeyrierasN

2007 Identification of the zebrafish IFN receptor: implications for the origin of the vertebrate IFN system. J Immunol 178 4385 4394

7. Lopez-MunozA

RocaFJ

SepulcreMP

MeseguerJ

MuleroV

2010 Zebrafish larvae are unable to mount a protective antiviral response against waterborne infection by spring viremia of carp virus. Dev Comp Immunol 34 546 552

8. SandersG

BattsW

WintonJ

2003 Susceptibility of zebrafish (Danio rerio) to a model pathogen, spring viremia of carp virus. Comp Med 53 514 521

9. PhelanP

PressleyM

WittenP

MellonM

BlakeS

2005 Characterization of snakehead rhabdovirus infection in zebrafish (Danio rerio). J Virol 79 1842 1852

10. LaPatraSE

BaroneL

JonesGR

ZonLI

2000 Effects of Infectious Hematopoietic Necrosis Virus and Infectious Pancreatic Necrosis Virus on hematopoietic precursors of the zebrafish. Blood Cells Mol Dis 26 445 452

11. AggadD

MazelM

BoudinotP

MogensenKE

HammingOJ

2009 The two groups of zebrafish virus-induced interferons signal via distinct receptors with specific and shared chains. J Immunol 183 3924 3931

12. NovoaB

RomeroA

MuleroV

RodriguezI

FernandezI

2006 Zebrafish (Danio rerio) as a model for the study of vaccination against viral haemorrhagic septicemia virus (VHSV). Vaccine 24 5806 5816

13. LuMW

ChaoYM

GuoTC

SantiN

EvensenO

2008 The interferon response is involved in nervous necrosis virus acute and persistent infection in zebrafish infection model. Mol Immunol 45 1146 1152

14. XuX

ZhangL

WengS

HuangZ

LuJ

2008 A zebrafish (Danio rerio) model of infectious spleen and kidney necrosis virus (ISKNV) infection. Virology 376 1 12

15. SpicklerAR

2007 Infectious Hematopoietic Necrosis. Center for Food Security and Public Health, Iowa State University

16. AmendD

YasutakeW

MeadR

1969 A hematopoietic virus of rainbow trout and sockeye. Trans Am Fish Soc 98 796 804

17. DroletB

RohovecJ

LeongJ

1994 The route of entry and progression of infectious haematopoietic necrosis virus in Oncorhyncus mykiss (Walbaum): a sequential immunohistochemical study. J Fish Dis 17 337 347

18. HarmacheA

LeBerreM

DroineauS

GiovanniniM

BrémontM

2006 Bioluminescence imaging of live infected salmonids reveals that the fin bases are the major portal entry of novirhabdoviruses. J Virol 80 3655 3659

19. McHenryMJ

LauderGV

2005 The mechanical scaling of coasting in zebrafish (Danio rerio). J Exp Biol 208 2289 2301

20. BanerjeeAK

1987 Transcription and replication of rhabdoviruses. Microbiol Rev 51 66 87

21. IsogaiS

HoriguchiM

WeinsteinBM

2001 The vascular anatomy of the developing zebrafish: an atlas of embryonic and early larval development. Dev Biol 230 278 301

22. LawsonN

WeinsteinB

2002 In vivo imaging of embryonic vascular development using transgenic zebrafish. Dev Biol 248 307 318

23. TraverD

PawB

PossK

PenberthyW

LinS

2003 Transplantation and in vivo imaging of multilineage engraftment in zebrafish bloodless mutants. Nat Immunol 4 1238 1246

24. Schneider-SchauliesJ

MeulenV

Schneider-SchauliesS

2003 Measles infection of the central nervous system. J Neurovirol 9 247 252

25. AlexakiA

LiuY

WigdahlB

2008 Cellular reservoirs of HIV-1 and their role in viral persistence. Curr HIV Res 6 388 400

26. SeternesT

SorensenK

SmedsrodB

2002 Scavenger endothelial cells of vertebrates: a nonperipheral leukocyte system for high-capacity elimination of waste macromolecules. Proc Natl Acad Sci U S A 99 7594 7597

27. BearzottiM

DelmasB

LamoureuxA

LoustauAM

ChilmonczykS

1999 Fish rhabdovirus cell entry is mediated by fibronectin. J Virol 73 7703 7709

28. LiuX

CollodiP

2002 Novel form of fibronectin from zebrafish mediates infectious hematopoietic necrosis virus infection. J Virol 76 492 498

29. Chelbi-AlixMK

VidyA

El BougriniJ

BlondelD

2006 Rabies viral mechanisms to escape the IFN system: the viral protein P interferes with IRF-3, Stat1, and PML nuclear bodies. J Interferon Cytokine Res 26 271 280

30. WesterfieldM

1993 The zebrafish book. A guide for the laboratory use of zebrafish (Brachydanio rerio). Eugene The University of Oregon Press

31. LevraudJ

Colucci-GuyonE

ReddM

LutfallaG

HerbomelP

2008 In vivo analysis of zebrafish innate immunity.

EwbankJ

VivierE

Innate Immunity Totowa, CA Humana Press 337 363

32. LutfallaG

UzéG

2006 Performing quantitative RT-PCR experiments.

KimmelA

OliverB

DNA microarrays San Diego Elsevier Academic Press 383 397

33. McCurleyAT

CallardGV

2008 Characterization of housekeeping genes in zebrafish: male-female differences and effects of tissue type, developmental stage and chemical treatment. BMC Mol Biol 9 102

34. BiacchesiS

YuYX

BearzottiM

TafallaC

Fernandez-AlonsoM

2000 Rescue of synthetic salmonid rhabdovirus minigenomes. J Gen Virol 81 1941 1945

35. NovakAE

RiberaAB

2003 Immunocytochemistry as a tool for zebrafish developmental neurobiology. Methods Cell Sci 25 79 83

36. BiacchesiS

BearzottiM

BouguyonE

BremontM

2002 Heterologous exchanges of the glycoprotein and the matrix protein in a Novirhabdovirus. J Virol 76 2881 2889

Štítky
Hygiena a epidemiológia Infekčné lekárstvo Laboratórium

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