Immunological properties of attenuated variants of African swine fever virus isolated in the Russian Federation

Балышева, В. И.; Prudnikova, E. Yu.; Galnbek, T.V.; Балышев, В. М.

doi:10.3103/s1068367415020056

Cited by 13 publications

(18 citation statements)

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“…RU 2439152 C1. Balyshev et al., ); (ii) attenuated variants of ASFV strain Stavropol 2008 after 24 and 33 passages in the continuous cell line A 4 C 2 /9k (Stavropol 2008 A 4 C 2 /9k) with a virus titre of 6.0–6.5 log 10 HAD 50 /ml (Prudnikova, ; Balysheva et al., ); (iii) virulent ASFV strain Congo‐49 (К‐49) (genotype I, seroimmunotype II; isolated in Central Africa), deposited in the form of virus‐containing blood with a virus titre of 7.0–7.5 log 10 HAD 50 /ml; and (iv) attenuated ASFV strain КК‐262/C (genotype I, seroimmunotype II), obtained by adaptation of the strain K‐49 to the continuous porcine kidney cell line PPK‐66b and following selection in a primary culture of porcine bone marrow (PBM) based on ‘loose’ haemadsorption (virus titre of 6.0–6.5 log 10 HAD 50 /ml) (Pat. RU 2452511 C1.…”

Section: Methodsmentioning

confidence: 99%

Recombinant Protein p30 for Serological Diagnosis of African Swine Fever by Immunoblotting Assay

Kazakova

Imatdinov

Dubrovskaya

et al. 2016

Transbound Emerg Dis

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This article is devoted to the development and evaluation of the immunoblotting test system for serological diagnosis of African swine fever (ASF), based on the highly purified recombinant p30 of ASF virus (ASFV) strain Stavropol 01/08 (Stavropol 2008), representative of the ASFV currently circulating in the Russian Federation. The main project stages are as follows: (i) cloning of the central hydrophilic region of the ASFV gene CP204L (p30) into a prokaryotic vector; (ii) expression and chromatographic purification of the recombinant product p30 with thioredoxin and poly-histidine site (p30e1_TrxA_6xHis); (iii) development of the immunoblotting test system (Rec p30-IB) using the highly purified recombinant p30; and (iv) evaluation of Rec p30-IB using sera and organ samples from domestic pigs and wild boars experimentally or naturally infected by ASFV. Testing of the Rec p30-IB showed the diagnostic specificity and sensitivity of the assay to be 98.75% and 100.00%, respectively. High sensitivity of the Rec p30-IB allowed the detection of ASFV-specific antibodies in samples of organs of the immune system and blood sera, collected from domestic pigs and wild boars, starting from 6 to 8 days post-infection, regardless of virus virulence, seroimmunotype and geographic origin of the samples (East Europe, South Europe, West Europe, Central and south-east Africa).

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Section: Methodsmentioning

confidence: 99%

Recombinant Protein p30 for Serological Diagnosis of African Swine Fever by Immunoblotting Assay

Kazakova

Imatdinov

Dubrovskaya

et al. 2016

Transbound Emerg Dis

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“…No cytopathic effects were observed during 20 passages in CV1, although virus presence in these cells was detected by immunofluorescence. When pigs were inoculated with the cultural virus of CV1 passage 20, it was observed that the virus lost pathogenicity; nevertheless, it was unable to confer protection against challenge with a virulent parental strain [ 93 ].…”

Section: Monkey-derived Continuous Cell Linesmentioning

confidence: 99%

“…A 4 C 2 and A 4 C 2 /9k are hybrid cell lines of SPEV TK with swine lymphocytes and were used to adapt the Russian ASFV strain Stavropol 01/08. Both epitheliocytes and lymphocyte-like cells were present in A 4 C 2 , whereas A 4 C 2 /9k presented an increased content of lymphocyte-like cells [ 93 ]. Adaptation was performed with several passage methods.…”

Section: Porcine Continuous Cell Linesmentioning

confidence: 99%

“…Other isolates that multiplied in this cell line were Sveromorsk 2010, Volgograd/Kalach 2012, Tver/Zavidovo 2012, etc. Nevertheless, the virus titer determined in the A 4 C 2 /9k cell culture was 0.7–1.25 log lower than that in the peripheral blood monocyte cell culture [ 93 ]. Interestingly, in both cell lines, hemadsorption was visible after adding 0.5% suspension of pig erythrocytes [ 93 ].…”

Section: Porcine Continuous Cell Linesmentioning

confidence: 99%

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Cell Lines for the Development of African Swine Fever Virus Vaccine Candidates: An Update

2022

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African swine fever virus (ASFV) is the etiological agent of a highly lethal disease in both domestic and wild pigs. The virus has rapidly spread worldwide and has no available licensed vaccine. An obstacle to the construction of a safe and efficient vaccine is the lack of a suitable cell line for ASFV isolation and propagation. Macrophages are the main targets for ASFV, and they have been widely used to study virus–host interactions; nevertheless, obtaining these cells is time-consuming and expensive, and they are not ethically suitable for the production of large-scale vaccines. To overcome these issues, different virulent field isolates have been adapted on monkey or human continuous cells lines; however, several culture passages often lead to significant genetic modifications and the loss of immunogenicity of the adapted strain. Thus, several groups have attempted to establish a porcine cell line able to sustain ASFV growth. Preliminary data suggested that some porcine continuous cell lines might be an alternative to primary macrophages for ASFV research and for large-scale vaccine production, although further studies are still needed. In this review, we summarize the research to investigate the most suitable cell line for ASFV isolation and propagation.

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“…Using a similar strategy, the virulent ASFV strain Stavropol 01/08 was attenuated by passages in a hybrid cell line A 4 C 2 /9k. The ASFV of passages 24 and 33 in A 4 C 2 /9k cells and passage 20 in CV-1 cells lost pathogenicity to pigs but did not protect pigs against virulent strain infection [ 15 ]. The virulent genotype II ASFV Georgia strain (ASFV-G) lost pathogenicity and immunogenicity after 110 passages in Vero cells [ 16 ].…”

Section: Strategies For Generating Asf Lav Candidatesmentioning

confidence: 99%

Current efforts towards safe and effective live attenuated vaccines against African swine fever: challenges and prospects

et al. 2021

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Background African swine fever (ASF) is a fatal hemorrhagic disease in domestic pigs and wild boar caused by African swine fever virus (ASFV). Since ASF has been introduced into Europe and Asia, the major pig-raising areas, posing a huge threat to the pork industry worldwide. Currently, prevention and control of ASF are basically dependent on strict biosecurity measures and stamping-out policy once ASF occurs. Main text The major risks of ASF spread are insufficient biosecurity measures and human behaviors. Therefore, a safe and effective vaccine seems to be a reasonable demand for the prevention and control of ASF. Due to the efficacy advantage over other types of vaccines, live attenuated vaccines (LAVs), especially virulence-associated genes deleted vaccines, are likely to be put into emergency and conditional use in restricted areas if ASF is out of control in a country with a huge pig population and pork consumption, like China. However, the safety, efficacy, and genetic stability of current candidate ASF LAVs require comprehensive clinical evaluations prior to country-wide field application. Several critical issues need to be addressed to commercialize an ideal ASF LAV, including a stable cell line for manufacturing vaccines, differentiation of infected from vaccinated animals (DIVA), and cross-protection from different genotypes. Conclusion A safe and effective DIVA vaccine and an accompanying diagnostic assay will facilitate the prevention, control, and eradication of ASF, which is quite challenging in the near future. Graphical Abstract

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Immunological properties of attenuated variants of African swine fever virus isolated in the Russian Federation

Cited by 13 publications

References 1 publication

Recombinant Protein p30 for Serological Diagnosis of African Swine Fever by Immunoblotting Assay

Recombinant Protein p30 for Serological Diagnosis of African Swine Fever by Immunoblotting Assay

Cell Lines for the Development of African Swine Fever Virus Vaccine Candidates: An Update

Current efforts towards safe and effective live attenuated vaccines against African swine fever: challenges and prospects

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