Sel’skokhozyaistvennaya Biologiya [Agricultural Biology], 2018, Vol. 53, ¹ 4, p. 860-867.
(how to cite this article)

doi: 10.15389/agrobiology.2018.4.860eng

UDC 636.4:619:578:[577.2.08+51-76

Acknowledgements:

Supported financially by Russian Science Foundation (the research project “Design of African swine fever virus candidate vaccine based on chimeric viruses” No 16-16-00090)

 

THE STUDY OF ANTIGENICITY, IMMUNOGENICITY AND PROTECTIVE POTENTIAL OF DNA CONSTRUCTS CONTAINING FRAGMENTS OF GENES CP204L, E183L OR EP402R OF AFRICAN SWINE FEVER VIRUS STRAIN MK-200

A.R. Imatdinov, O.A. Dubrovskaya, D.Yu. Morozova, V.M. Lyska,
A.D. Sereda

Federal Research Center for Virology and Microbiology, Federal Agency of Scientific Organizations, 1, ul. Akademika Bakuleva, pos. Vol’ginskii, Petushinskii Region, Vladimir Province, 601125 Russia, e-mail almazlcf@yandex.ru, olgadubrovskaya@list.ru, lady_d.morozova@mail.ru, diagnoz3@yandex.ru, sereda-56@mail.ru (✉ corresponding author)

ORCID:
Imatdinov A.R. orcid.org/0000-0003-2889-6112
Lyska V.M. orcid.org/0000-0001-5302-3108
Dubrovskaya O.A. orcid.org/0000-0002-3168-7947
Sereda A.D. orcid.org/0000-0001-8300-5234
Morozova D.Yu. orcid.org/0000-0001-5486-9981
The authors declare no conflict of interests

Received April 16, 2018

 

African swine fever (ASF) is a viral, contagious and septic disease affecting wild and domestic pigs of all breeds and age groups. In both domestic pigs and wild European boars affected, some hyperacute or acute forms of the infection are observed which are characterized by fever, signs of toxicosis, hemorrhagic diathesis with mortality rates of up to 100 %. In endemic regions (e.g., some countries of East Africa), a subacute form of the disease with a mortality of 30 to 70 %, as well as a chronic one with very low mortality levels are reported (S. Blome et al., 2013; C. Gabriel et al., 2011; JM Sánchez -Vizcaíno et al., 2015). No vaccine against African swine fever is currently available, and the research works aimed at the development of live, inactivated and subunit vaccines have not achieved the intended result yet (P.J. Sánchez-Cordón et al., 2017; V. O’Donnell et al., 2016; S. Blome et al., 2014). Nevertheless, the opportunity of obtaining a DNA vaccine using the genes of potentially protective ASFV proteins p30, p54 and/or CD2v is considered to be a promising option in a number of laboratories worldwide. The proteins p30 and p54 are functionally important in attaching the ASF virus to the target cell. The antibody to p54 blocks the virion binding to the macrophage, whereas the antibody to p30 inhibits the virion penetration into the cell. The protein CD2v determines the hemadsorbing properties of the virus (S.D. Kollnberger et al., 2002; M.G. Barderas et al., 2001; J.G. Neilan et al., 2004). This work has been performed to study effects of the translation products of recombinant plasmids pCI-neo/ASFV/p30, pCI-neo/ASFV/p54 and pCI-neo/ASFV/CD2v induced in adhesive cells (A-cells) after transfection. The antigenic activity of the recombinant proteins produced with these DNA constructs was compared in permanent cell line HEK-293T and swine leukocyte (SL) autologous primary cultures using a direct fluorescence technique. The highest expression of the antigen-active translation products in HEK-293T and SL cells transfected with pCI-neo/ASFV/p30, pCI-neo/ASFV/p54 or pCI-neo/ASFV/CD2v was observed on day 2. The peculiarity of the pig immunization schedule applied was that the animals were thrice immunized at a 14-day interval with autologous LS A cells transfected in vitro with the above recombinant plasmids. For this, as much as 90 cm3 of LC cell culture was produced using blood samples from each of the animals No.No. 1-4. On day 2 of the culturing, 90 μg of pCI-neo/ASFV/p30 (No. 1), pCI-neo/ASFV/p54 (No. 2) or pCI-neo/ASFV/CD2v (No. 3) was added thereto. The LS culture obtained from the pig No. 4 in a volume of 90 cm3 was divided into three portions of 30 cm3, and each one was transfected with one of the three constructs (i.e., pCI-neo/ASFV/p30, pCI-neo/ASFV/p54 or pCI-neo/ASFV/CD2v) by adding 30 μg of the plasmid DNA. Two days later, the pigs No. 1 to No. 4 were inoculated into the central auricular vein with about 107 autologous transfected A-cells of LS cultures. On days 14, 28 and 42, no antibody against ASFV proteins was detected in the blood of the immunized pigs using indirect solid-phase ELISA and immunoblotting. After the pigs were infected into the neck with 102 HAU50 of an ASFV strain Mozambique-78 on day 42, the four pre-immunized pigs (No.No. 1-4) died on day 6 to 8 and the control one died (No. 5) on day 13. Thus, the immunization of pigs with the autologous and antigenically active LS cells transfected with the recombinant plasmids pCI-neo/ASFV/p30, pCI-neo/ASFV/p54 or pCI-neo/ASFV/CD2v failed to provide antibody response or protection against the challenge.

Keywords: African swine fever, ASFV, recombinant plasmids, ASFV proteins ð30, ð54 and CD2v, antigenicity, immunogenicity.

 

Full article (Rus)

Full article (Eng)

 

REFERENCES

  1. Blome S., Gabriel C., Beer M. Pathogenesis of African swine fever in domestic pigs and European wild boar. Virus Res., 2013, 173(1): 122-130 CrossRef
  2. Gabriel C., Blome S., Malogolovkin A., Parilov S., Kolbasov D., Teifke J.P., Beer M. Characterization of African swine fever virus Caucasus isolate in European wild boars. Emerg. Infect. Dis., 2011, 17(12): 2342-2345 CrossRef
  3. Sánchez-Vizcaíno J.M., Mur L., Gomez-Villamandos J.C., Carrasco L. An update on the epidemiology and pathology of African swine fever. J. Comp. Pathol., 2015, 152(1): 9-21 CrossRef
  4. Sánchez-Cordón P.J., Chapman D., Jabbar T., Reis A.L., Goatley L., Netherton C.L., Taylor G., Montoya M., Dixon L. Different routes and doses influence protection in pigs immunised with the naturally attenuated African swine fever virus isolate OURT88/3. Antiviral Res., 2017, 138: 1-8 CrossRef
  5. O’Donnell V., Holinka L.G., Sanford B., Krug P.W., Carlson J., Pacheco J.M., Reese B., Risatti G.R., Gladue D.P., Borca M.V. African swine fever virus Georgia isolate harboring deletions of 9GL and MGF360/505 genes is highly attenuated in swine but does not confer protection against parental virus challenge. Virus Res., 2016, 221: 8-14 CrossRef
  6. Blome S., Gabriel C., Beer M. Modern adjuvants do not enhance the efficacy of an inactivated African swine fever virus vaccine preparation. Vaccine, 2014, 32(31): 3879-3882 CrossRef
  7. Lacasta A., Ballester M., Monteagudo P.L., Rodríguez J.M., Salas M.L., Accensi F., Pina-Pedrero S., Bensaid A., Argilaguet J., López-Soria S., Hutet E., Le Potier M.F., Rodríguez F. Expression library immunization can confer protection against lethal challenge with African swine fever virus. J. Virol., 2014, 88(22): 13322-13332 CrossRef
  8. Argilaguet J.M., Pérez-Martín E., Nofrarías M., Gallardo C., Accensi F., Lacasta A., Mora M., Ballester M., Galindo-Cardiel I., López-Soria S., Escribano J.M., Reche P.A., Rodríguez F. DNA vaccination partially protects against African swine fever virus lethal challenge in the absence of antibodies. PLoS ONE, 2012, 7(9): e40942 CrossRef
  9. Sereda A.D., Kazakova A.S., Imatdinov A.R., Kolbasov D.V. Humoral and cell immune mechanisms under African swine fever (review). Sel’skokhozyaistvennaya Biologiya [Agricultural Biology], 2015, 50(6): 709-718 CrossRef
  10. Lacasta A., Monteagudo P.L., Jiménez-Marín Á., Accensi F., Ballester M., Argilaguet J., Galindo-Cardiel I., Segalés J., Salas M.L., Domínguez J., Moreno Á., Garrido J.J., Rodríguez F. Live attenuated African swine fever viruses as ideal tools to dissect the mechanisms involved in viral pathogenesis and immune protection. Vet. Res., 2015, 46: 135 CrossRef
  11. Takamatsu H.H., Denyer M.S., Lacasta A., Stirling C.M., Argilaguet J.M., Netherton C.L., Oura C.A., Martins C., Rodríguez F. Cellular immunity in ASFV responses. Virus Res., 2013, 173(1): 110-121 CrossRef
  12. Kollnberger S.D., Gutierrez-Castañeda B., Foster-Cuevas M., Corteyn A., Parkhouses R.M.E. Identification of the principal serological immunodeterminants of African swine fever virus by screening a virus cDNA library with antibody. J. Gen. Virol., 2002, 83: 1331-1342 CrossRef
  13. Barderas M.G., Rodriguez F., Gomez-Puertas P., Aviles M., Beitia F., Alonso C., Escribano J.M. Antigenic and immunogenic properties of a chimera of two immunodominant African swine fever virus proteins. Arch. Virol., 2001, 146: 1681-1691 CrossRef
  14. Neilan J.G., Zsak L., Lu Z., Burrage T.G., Kutish G.F., Rock D.L. Neutralizing antibodies to African swine fever virus proteins p30, p54, and p72 are not sufficient for antibody-mediated protection. Virology, 2004, 319: 337-342 CrossRef
  15. Ruiz-Gonzalvo F., Rodríguez F., Escribano J.M. Functional and immunological properties of the baculovirus-expressed hemagglutinin of African swine fever virus. Virology, 1996, 218: 285-289 CrossRef
  16. Takamatsu H.H., Denyer M.S., Lacasta A., Stirling C.M., Argilaguet J.M., Netherton C.L., Oura C.A., Martins C., Rodríguez F. Cellular immunity in ASFV responses. Virus Res., 2013, 173(1): 110-121 CrossRef
  17. Argilaguet J.M., Pérez-Martín E., López S., Goethe M., Escribano J.M., Giesow K., Keil G.M., Rodríguez F. BacMam immunization partially protects pigs against sublethal challenge with African swine fever virus. Antiviral Res., 2013, 98(1): 61-65 CrossRef
  18. Imatdinov A.R., Sereda A.D., Imatdinov I.R., Kazakova A.S., Dubrovskaya O.A., Kolbasov D.V. Expression of recombinant genes encoding fragments of the protective important proteins of African swine fever virus in eucaryotic cells. Sel’skokhozyaistvennaya Biologiya [Agricultural Biology], 2016, 51(6): 837-844 CrossRef
  19. Gómez-Puertas P., Rodriguez F., Oviedo J.M., Brun A., Alonso C., Escribano J.M. The African swine fever virus proteins p54 and p30 are involved in two distinct steps of virus attachment and both contribute to the antibody-mediated protective immune response. Virology, 1998, 243: 461-471 CrossRef
  20. Rodriguez F., Alcaraz S., Yanez R.J., Rodriguez J.M., Alonso C., Rodriguez J.F., Escribano J.M. Characterization and molecular basis of heterogeneity of the African swine fever virus envelope protein p54. J. Virol., 1994, 68(11): 7244-7252.
  21. Goatley L.C., Dixon L.K. Processing and Localization of the African swine fever virus CD2v transmembrane protein. J. Virol., 2011, 85(7): 3294-3305 CrossRef
  22. Malogolovkin A., Burmakina G., Titov I., Sereda A., Gogin A., Baryshnikova E., Kolbasov D. Comparative analysis of African swine fever virus genotypes and serogroups. Emerg. Infect. Dis., 2015, 21(2): 312-315 CrossRef
  23. Strizhakova O.M., Lyska V.M., Malogolovkin A.S., Novikova M.B., Sidlik M.V., Nogina I.V., Shkaev A.E., Balashova E.A., Kurinnov V.V., Vasil'ev A.P. Validation of an ELISA kit for detection of antibodies against ASF virus in blood or spleen of domestic pigs and wild boars. Sel’skokhozyaistvennaya Biologiya [Agricultural Biology], 2016, 51(6): 845-852 CrossRef
  24. Immunoblotting OIE for serological diagnosis of African swine fever (SOP/CISA/ASF/IB/1/2008). Available http://asf-referencelab.info/asf/images/files/SOPs/SOP-AFSIB12008.pdf. No date.
  25. Ryzhova E.V., Belyanin S.A., Kolbasov D.V., Balyshev V.M., Kurinnov V.V., Pronin V.V., Korneva G.V. Rossiiskii veterinarnyi zhurnal, 2012, 1: 10-14 (in Russ.).
Jancovich J.K., Chapman D., Hansen D.T., Robida M.D., Loskutov A., Craciunescu F.,  Borovkov A.,  Kibler K.,  Goatley L., King K.,  Netherton C,L.,  Taylor G., Jacobs B.,  Sykes K., Dixon L.K. Immunization of pigs by DNA prime and recombinant vaccinia virus boost to identify and rank African swine fever virus immunogenic and protective proteins. J. Virol., 2018, 92(8): e02219-17 CrossRef

 

back