African swine fever virus DNA: deletions and additions during adaptation to growth in monkey kidney cells

Tabarés, E.; Olivares, Isabel; Santurde, G.; García, Miguel Martínez; Martín, Elena; Carnero, M. E.

doi:10.1007/bf01314431

Cited by 59 publications

(58 citation statements)

References 33 publications

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“…The process of ASFV adaptation to be able to replicate in established cell lines is usually accompanied by attenuation of virulence and significant modifications of the viral genome, generally affecting the variable genomic ends (21,22,23,24,25). Unfortunately, in most of these cases, detailed descriptions of the adaptation process and the associated genetic and phenotypic changes of ASFV are not readily available.…”

Section: Discussionmentioning

confidence: 99%

“…Adaptation of virulent field isolates to replicate in established cell lines from different species has been attempted, and the process has led to decreased virulence of adapted viruses in swine (7,21,22). The process of ASFV adaptation to be able to replicate in established cell lines is usually accompanied by attenuation of virulence and significant modifications of the viral genome, generally affecting the variable genomic ends (21,22,23,24,25).…”

Section: Discussionmentioning

confidence: 99%

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The Progressive Adaptation of a Georgian Isolate of African Swine Fever Virus to Vero Cells Leads to a Gradual Attenuation of Virulence in Swine Corresponding to Major Modifications of the Viral Genome

Krug

Holinka

O’Donnell

et al. 2015

J Virol

148

159

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African swine fever virus (ASFV) causes a contagious and often lethal disease of feral and domestic swine. Experimental vaccines derived from naturally occurring, genetically modified, or cell culture-adapted ASFV have been evaluated, but no commercial vaccine is available to control African swine fever (ASF). We report here the genotypic and phenotypic analysis of viruses obtained at different passages during the process of adaptation of a virulent ASFV field isolate from the Republic of Georgia (ASFV-G) to grow in cultured cell lines. ASFV-G was successively passaged 110 times in Vero cells. Viruses obtained at passages 30, 60, 80, and 110 were evaluated in vitro for the ability to replicate in Vero cells and primary swine macrophages cultures and in vivo for assessing virulence in swine. Replication of ASFV-G in Vero cells increased with successive passages, corresponding to a decreased replication in primary swine macrophages cultures. In vivo, progressive loss of virus virulence was observed with increased passages in Vero cells, and complete attenuation of ASFV-G was observed at passage 110. Infection of swine with the fully attenuated virus did not confer protection against challenge with virulent parental ASFV-G. Full-length sequence analysis of each of these viruses revealed significant deletions that gradually accumulated in specific areas at the right and left variable ends of the genome. Mutations that result in amino acid substitutions and frameshift mutations were also observed, though in a rather limited number of genes. The potential importance of these genetic changes in virus adaptation/attenuation is discussed. IMPORTANCEThe main problem in controlling ASF is the lack of vaccines. Attempts to produce vaccines by adaptation of ASFV to cultured cell lines have been made. These attempts led to the production of attenuated viruses that conferred only homologous protection. Specifics regarding adaptation of these isolates to cell cultures have been insufficiently described. Details like the numbers of passages required to obtain attenuated viruses, genetic modifications introduced into the virus genomes along passages, and the extent of attenuation and induced protective efficacy are not readily available. In this study, we assessed the changes that lead to decreased growth in swine macrophages and to attenuation in swine. Loss of virulence, probably associated with limited replication in vivo, may lead to the lack of protective immunity in swine observed after challenge. This report provides valuable information that can be used to further the understanding of ASFV gene function, virus attenuation, and protection against infection.A frican swine fever (ASF) is a contagious disease of swine caused by ASF virus (ASFV), an enveloped virus containing a large (190-kbp) double-stranded DNA (dsDNA) genome. ASFV infection of domestic pigs can induce a spectrum of disease severity, from highly lethal to subclinical, depending on host characteristics and the particular circulating virus strain (1).A...

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

The Progressive Adaptation of a Georgian Isolate of African Swine Fever Virus to Vero Cells Leads to a Gradual Attenuation of Virulence in Swine Corresponding to Major Modifications of the Viral Genome

Krug

Holinka

O’Donnell

et al. 2015

J Virol

148

159

View full text Add to dashboard Cite

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“…ASFV variable regions comprise the left 35-kb and the right 15-kb ends of the genome and contain at least five multigene families (MGFs): MGF 100, MGF 110, MGF 300, MGF 360, and MGF 530 (3,4,15,27,61,64). Variations in these regions, including gene deletion events, are observed during ASFV adaptation to monkey cell lines (6,58). Recently, MGF 360 and MGF 530 genes in the left variable region of the ASFV genome have been shown to function in macrophage host range (65).…”

mentioning

confidence: 99%

Novel Swine Virulence Determinant in the Left Variable Region of the African Swine Fever Virus Genome

Neilan

Zsák

et al. 2002

J Virol

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Previously we have shown that the African swine fever virus (ASFV) NL gene deletion mutant E70⌬NL is attenuated in pigs. Our recent observations that NL gene deletion mutants of two additional pathogenic ASFV isolates, Malawi Lil-20/1 and Pr4, remained highly virulent in swine (100% mortality) suggested that these isolates encoded an additional virulence determinant(s) that was absent from E70. To map this putative virulence determinant, in vivo marker rescue experiments were performed by inoculating swine with infectiontransfection lysates containing E70 NL deletion mutant virus (

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“…It is the sole member of the newly named Asfarviridae and the only known DNA arbovirus (10). ASFV is a large icosahedral virus which contains a linear doublestranded DNA genome (170 to 190 kbp) encoding approximately 165 viral proteins (8,29).…”

mentioning

confidence: 99%

Preclinical Diagnosis of African Swine Fever in Contact-Exposed Swine by a Real-Time PCR Assay

et al. 2005

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A fluorogenic probe hydrolysis (TaqMan) PCR assay for African swine fever virus (ASFV) was developed and evaluated in experimentally infected swine. This sensitive and specific one-step single-tube assay, which can be performed in 2 h or less, detected viral DNA in tonsil scraping samples 2 to 4 days prior to onset of clinical disease. Thus, the assay would have application for preclinical diagnosis of African swine fever and surveillance and/or emergency management of a disease outbreak

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African swine fever virus DNA: deletions and additions during adaptation to growth in monkey kidney cells

Cited by 59 publications

References 33 publications

The Progressive Adaptation of a Georgian Isolate of African Swine Fever Virus to Vero Cells Leads to a Gradual Attenuation of Virulence in Swine Corresponding to Major Modifications of the Viral Genome

The Progressive Adaptation of a Georgian Isolate of African Swine Fever Virus to Vero Cells Leads to a Gradual Attenuation of Virulence in Swine Corresponding to Major Modifications of the Viral Genome

Novel Swine Virulence Determinant in the Left Variable Region of the African Swine Fever Virus Genome

Preclinical Diagnosis of African Swine Fever in Contact-Exposed Swine by a Real-Time PCR Assay

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