Resistance to African swine fever virus among African domestic pigs appears to be associated with a distinct polymorphic signature in the RelA gene and upregulation of RelA transcription.


Journal

Virology journal
ISSN: 1743-422X
Titre abrégé: Virol J
Pays: England
ID NLM: 101231645

Informations de publication

Date de publication:
24 Apr 2024
Historique:
received: 07 01 2024
accepted: 24 03 2024
medline: 25 4 2024
pubmed: 25 4 2024
entrez: 24 4 2024
Statut: epublish

Résumé

African swine fever virus (ASFV) is a highly contagious and fatal hemorrhagic disease of domestic pigs, which poses a major threat to the swine industry worldwide. Studies have shown that indigenous African pigs tolerate ASFV infection better than European pigs. The porcine v-rel avian reticuloendotheliosis viral oncogene homolog A (RelA) encoding a p65 kD protein, a major subunit of the NF-kB transcription factor, plays important roles in controlling both innate and adaptive immunity during infection with ASFV. In the present study, RelA genes from ASFV-surviving and symptomatic pigs were sequenced and found to contain polymorphisms revealing two discrete RelA amino acid sequences. One was found in the surviving pigs, and the other in symptomatic pigs. In total, 16 nonsynonymous SNPs (nsSNPs) resulting in codon changes were identified using bioinformatics software (SIFT and Polyphen v2) and web-based tools (MutPre and PredictSNP). Seven nsSNPs (P374-S, T448-S, P462-R, V464-P, Q478-H, L495-E, and P499-Q) were predicted to alter RelA protein function and stability, while 5 of these (P374-S, T448-S, P462-R, L495-E, and Q499-P) were predicted as disease-related SNPs.Additionally, the inflammatory cytokine levels of IFN-α, IL-10, and TNF-α at both the protein and the mRNA transcript levels were measured using ELISA and Real-Time PCR, respectively. The resulting data was used in correlation analysis to assess the association between cytokine levels and the RelA gene expression. Higher levels of IFN-α and detectable levels of IL-10 protein and RelA mRNA were observed in surviving pigs compared to healthy (non-infected). A positive correlation of IFN-α cytokine levels with RelA mRNA expression was also obtained. In conclusion, 7 polymorphic events in the coding region of the RelA gene may contribute to the tolerance of ASFV in pigs.

Identifiants

pubmed: 38658979
doi: 10.1186/s12985-024-02351-9
pii: 10.1186/s12985-024-02351-9
doi:

Substances chimiques

Transcription Factor RelA 0

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

93

Informations de copyright

© 2024. The Author(s).

Références

Montgomery RE. On a form of swine fever occurring in British East Africa (Kenya Colony). J Compar Pathol. 1921;4:159–91.
doi: 10.1016/S0368-1742(21)80031-4
Dixon LK, Chapman DA, Netherton CL, Upton C. African swine fever virus replication and genomics. Virus Res. 2013;173(1):3–14. https://doi.org/10.1016/j.virusres.2012.10.020
doi: 10.1016/j.virusres.2012.10.020 pubmed: 23142553
Penrith ML, Thomson GR, Bastos ADS. African swine fever. In: Coetzer JAW, Tustin RC, editors. Infectious diseases of livestock. Volume 2, 2nd ed. Oxford, United Kingdom: Oxford University Press; 2004:1088–119.
Mulumba–Mfumu LK, Achenbach EJ, Mauldin MR, Dixon LK, Tshilenge CG, Thiry E, Moreno N, Blanco E, Saegerman C, Lamien CE, Diallo A. Genetic assessment of African swine fever isolates involved in outbreaks in the Democratic Republic of Congo between 2005 and 2012 reveals co-circulation of p72 genotypes I, IX and XIV, including 19 variants. Viruses. 2017;9:31. https://doi.org/10.3390/v9020031
doi: 10.3390/v9020031 pubmed: 28218698 pmcid: 5332950
Bisimwa NP, Eunice MM, Joshua AO, Dedan G, Gedéon B, Charles M, et al. Evidence for the presence of African swine fever virus in apparently healthy pigs in South Kivu province of the Democratic Republic of Congo. Vet Microbiol. 2019;240:108521.
Bisimwa NP, Ongus JR, Tiambo CK, Machuka ME, Bisimwa BE, Steinaa L, Pelle R. First detection of African swine fever (ASF) virus genotype X and serogroup 7 in symptomatic pigs in the Democratic Republic of Congo. Virol J. 2020;17:135. https://doi.org/10.1186/s12985-020-01398
doi: 10.1186/s12985-020-01398 pubmed: 32883295 pmcid: 7468181
Anderson EC, Hutchings GH, Mukarati N, Wilkinson PJ. African swine fever virus infection of the bushpig (Potamochoerus porcus) and its significance in the epidemiology of the disease. Vet Microbiol. 1998;62:1–15. https://doi.org/10.1016/s0378-1135(98)00187-4
doi: 10.1016/s0378-1135(98)00187-4 pubmed: 9659687
Katale BZ, Fyumagwa RD, Mdaki ML, Hoare R. Prevalence of African swine fever virus in warthogs in the serengeti ecosytem. Tanzan Res Opin Anim Vet Sci. 2012;2(5):339–43.
Jori F: and, Bastos ADS. Role of wild suids in the epidemiology of African swine fever. EcoHealth. 2009;6:296–300.
doi: 10.1007/s10393-009-0248-7 pubmed: 19915917
Palgrave CJ, Gilmour L, Lowden CS, Lillico SG, Mellencamp MA, Whitelaw CB. Species-specific variation in RELA underlies differences in NF-κB activity: a potential role in African swine fever pathogenesis. J Virol. 2011;85(12):6008–14.
doi: 10.1128/JVI.00331-11 pubmed: 21450812 pmcid: 3126315
Uttenthal A, Braae UC, Ngowi HA, Rasmussen TB, Nielsen J, Johansen MV. ASFV in Tanzania: asymptomatic pigs harbor virus of molecular similarity to Georgia 2007. Vet Microbiol. 2013;165(1–2):173–6. https://doi.org/10.1016/j.vetmic.2013.01.003
doi: 10.1016/j.vetmic.2013.01.003 pubmed: 23398669
Abworo EO, Onzere C, Amimo JO, Riitho V, Mwangi W, Davies J, et al. Detection of African swine fever virus in the tissues of asymptomatic pigs in smallholder farming systems along the Kenya–Uganda border: implications for transmission inendemic areas and ASF surveillance in East Africa. J Gener Virol. 2017;98:1806–DOI1814. https://doi.org/10.1099/jgv.0.000848
doi: 10.1099/jgv.0.000848
Salguero FJ. Comparative pathology and pathogenesis of African swine fever infection in swine. Front Vet Sci. 2020;7:282.
doi: 10.3389/fvets.2020.00282 pubmed: 32509811 pmcid: 7248413
Murtaugh MP, Baarsch MJ, Zhou Y, Scamurra RW, Lin G. Inflammatory cytokines in animal health and disease. Vet Immunol Immunopathol. 1996;54(1–4):45–55.
doi: 10.1016/S0165-2427(96)05698-X pubmed: 8988847
Golding JP, Goatley L, Goodbourn S, Dixon LK, Taylor G, Netherton CL. Sensitivity of African swine fever virus to type I interferon is linked to genes within multigene families 360 and 505. Virol. 2016;493:154–61. https://doi.org/10.1016/j.virol.2016.03.019
doi: 10.1016/j.virol.2016.03.019
Zhu JJ, Ramanathan P, Bishop EA, O’Donnell V, Gladue DP, Borca MV. Mechanisms of African swine fever virus pathogenesis and immune evasion inferred from gene expression changes in infected swine macrophages. PLoS ONE. 2019;14(11):e0223955. https://doi.org/10.1371/journal.pone.0223955
doi: 10.1371/journal.pone.0223955 pubmed: 31725732 pmcid: 6855437
García-Belmonte R, Pérez-Núñez D, Pittau M, Richt JA, Revilla Y. African Swine Fever Virus Armenia/07 Virulent Strain Controls Interferon Beta Production through the cGAS-STING Pathway. J Virol. 2019;93(12):e02298–18. https://doi.org/10.1128/JVI.02298-18
Granja AG, Nogal ML, Hurtado C, Del Aguila C, Carrascosa AL, Salas ML, et al. The viral protein A238L inhibits TNF-alpha expression through a CBP/p300 transcriptional coactivators pathway. J Immunol. 2006;176:451–62. https://doi.org/10.4049/jimmunol.176.1.451
doi: 10.4049/jimmunol.176.1.451 pubmed: 16365438
Oeckinghaus A, Sankar, Ghosh S. The NF-κB family of transcription factors and its regulation. Cold Spring Harb Perspect Biol. 2009;1(4):a000034. https://doi.org/10.1101/cshperspect.a000034
doi: 10.1101/cshperspect.a000034 pubmed: 20066092 pmcid: 2773619
Dakal TC, Kala D, Dhiman G, Yadav V, Krokhotin A, Dokholyan NV. Predicting the functional consequences of nonsynonymous single nucleotide polymorphisms in IL8 gene. Scient Rep. 2017;7:6525. https://doi.org/10.1038/s41598-017-06575-4
doi: 10.1038/s41598-017-06575-4
Afonso CL, Piccone ME, Zaffuto KM, Neilan J, Kutish GF, Lu Z, et al. African swine fever virus multigene family 360 and 530 genes affect host interferon response. J Virol. 2004;78:1858–64.
doi: 10.1128/JVI.78.4.1858-1864.2004 pubmed: 14747550 pmcid: 369441
Revilla Y, Callejo M, Rodríguez JM, Culebras E, Nogal ML, Salas ML, Viñuela E, Fresno M. Inhibition of nuclear factor kappaB activation by a virus-encoded IkappaB-like protein. J Biol Chem. 27;273(9):5405–11. https://doi.org/10.1074/jbc.273.9.5405
Granja AG, Sabina P, Salas ML, Fresno M, Revilla Y. Regulation of inducible nitric oxide synthase expression by viral A238L-mediated inhibition of p65/RelA acetylation and p300 transactivation. J Virol. 2006;80(21):10487–96. https://doi.org/10.1128/JVI.00862-06.
Gerritsen ME, Williams AJ, Neish AS, Moore S, Shi Y, Collins T. CREB-binding protein/p300 are transcriptional coactivators of p65. Proc Natl Acad Sci USA. 1997;94:2927–32.
doi: 10.1073/pnas.94.7.2927 pubmed: 9096323 pmcid: 20299
Miskin JE, Abrams CC, Dixon LK. African swine fever virus protein A238L interacts with the cellular phosphatase calcineurin via a binding domain similar to that of NFAT. J Virol. 2000;74:9412–20.
doi: 10.1128/JVI.74.20.9412-9420.2000 pubmed: 11000210 pmcid: 112370
Wang P, Li W, Yang Y, Cheng N, Zhang Y, Zhang N, Yin Y, Tong L, Li Z, Luo JA. Polypeptide inhibitor of calcineurin blocks the calcineurin-NFAT signalling pathway in vivo and in vitro. J Enzym Inhib Med Chem. 2022;37:202–210.
Chen H, Wang Z, Gao X et al. ASFV pD345L protein negatively regulates NF-κB signalling by inhibiting IKK kinase activity. Vet Res. 2022;53:32 (2022). https://doi.org/10.1186/s13567-022-01050-z
Chen X, Li L, Yang Z, Li M, Fan S, Shi L, Ren Z, Cao X, Zhang Y, Han S, Wan B, Qiu H, Zhang G, He W. 2023. The African swine fever virus I10L protein inhibits the NF-κB signaling pathway by targeting IKKβ. J Virol. 2023;97:e00569–23. https://doi.org/10.1128/jvi.00569-23
Rodríguez CI, Nogal ML, Carrascosa AL, Salas ML, Fresno M, Revilla Y. African swine fever virus IAP-like protein induces the activation of nuclear factor kappa B. J Virol. 2002;76(8):3936–42. https://doi.org/10.1128/jvi.76.8.3936-3942.2002
Sim NL, Kumar P, Hu J, Henikoff S, Schneider G, Ng PC. SIFT web server: predicting effects of amino acid substitutions on proteins. Nucleic Acids Res. 2012;40:452–7.
doi: 10.1093/nar/gks539
Adzhubei I, Jordan DM, Sunyaev SR. Predicting functional effect of human missense mutations using PolyPhen-2. Curr Protoc Hum Genet. 2013;07(Unit720). https://doi.org/10.1002/0471142905.hg0720s76
Bendl J, Stourac J, Salanda O, Pavelka A, Wieben ED, Zendulka J, et al. PredictSNP: robust and accurate consensus classifier for prediction of disease-related mutations. PLOS Computat Biol. 2014;10:e1003440.
doi: 10.1371/journal.pcbi.1003440
Pejaver V, Urresti J, Lugo-Martinez J, Pagel KA, Lin GN, Nam H, et al. MutPred2: inferring the molecular and phenotypic impact of amino acid variants. bioRxiv. 2017;134981. https://doi.org/10.1101/134981
Cheng J, Randal A, Baldi P. Prediction of protein stability changes for single-site mutations using support vector machines. Proteins. 2006;1(624):1125–32.
doi: 10.1002/prot.20810
Narayana SA, Valasala H, Kamma S. In silico evaluation of nonsynonymous single nucleotide polymorphisms in the ADIPOQ gene associated with diabetes, obesity, and inflammation. Avicenna J Med Biotechnol. 2015;7(3):121–7.
Gil S, Sepúlveda N, Albina E, Leitaó A, Martins C. The low-virulent African swine fever virus (ASFV/NH/P68) induces enhanced expression and production of relevant regulatory cytokines (IFNa, TNFa and IL12p40) on porcine macrophages in comparison to the highly virulent ASFV/L60. Arch Virol. 2008;153:1845–54. https://doi.org/10.1007/s00705-008-0196-5
doi: 10.1007/s00705-008-0196-5 pubmed: 18787755 pmcid: 2831186
Blancho G, Gianello P, Germana S, Baetscher M, Sachs DH, LeGuern C. Molecular identification of porcine interleukin 10: regulation of expression in a kidney allograft model. Proc Natl Acad Sci USA. 1995;28;92(7):2800–4. https://doi.org/10.1073/pnas.92.7.2800
Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2-DDCT method. Methods. 2001;25:402–8. https://doi.org/10.1006/meth.2001.1262
doi: 10.1006/meth.2001.1262 pubmed: 11846609
Schmitz ML, Baeuerle PA. The p65 subunit is responsible for the strong transcription activating potential of NF-kappa B. EMBO J. 1991;10:3805–17.
doi: 10.1002/j.1460-2075.1991.tb04950.x pubmed: 1935902 pmcid: 453117
Li H, Ma G, Gui D, Zhao S, Wang P, He K, et al. Characterization of the porcine p65 subunit of NF-κB and its association with virus antibody levels. Mol Immunol. 2011;48(6–7):914–23. https://doi.org/10.1016/j.molimm.2010.12.015
doi: 10.1016/j.molimm.2010.12.015 pubmed: 21269694
Law M, Corsino P, Parker NT, Law BK. Identification of a small molecule inhibitor of serine 276 phosphorylation of the p65 subunit of NF-kappaB using in silico molecular docking. Cancer Lett. 2010;291:217–24.
doi: 10.1016/j.canlet.2009.10.015 pubmed: 19910110
Stephen WG, Tait EB, Reid DR, Greaves TE, Wileman PP. Mechanism of inactivation of NF-kB by a viral homologue of IkBa. J Biol Cherm. 2000;275(44):34656–64.
doi: 10.1074/jbc.M000320200
Milanovic M, Kracht M, Schmitz ML. The cytokine-induced conformational switch of nuclear factor Kappab P65 is mediated by P65 phosphorylation. Biochem J. 2014;457:401–13. https://doi.org/10.1042/BJ20130780
doi: 10.1042/BJ20130780 pubmed: 24175631
Hoffmann A, Levchenko A, Scott ML, Baltimore D. The IkappaB-NF-kappaB signaling module: temporal control and selective gene activation. Science. 2002;298:1241–5. https://doi.org/10.1126/science.1071914
doi: 10.1126/science.1071914 pubmed: 12424381
Barnes P, Karin M. Nuclear factor kappaB: a pivotal transcription factor in chronic inflammatory diseases. N Engl J Med. 1997;336(15):1066–71.
doi: 10.1056/NEJM199704103361506 pubmed: 9091804
Akira S, Uematsu S, Takeuchi O. Pathogen recognition and innate immunity. Cell. 2006;124(4):783–801. https://doi.org/10.1016/j.cell.2006.02.015
doi: 10.1016/j.cell.2006.02.015 pubmed: 16497588
Brasier AR. The NF-κB Signaling Network: insights from systems approaches. In: Brasier AR, Lemon SM, Garcia-Sastre A, editors. Cellular Signaling and Innate Immune responses to RNA virus infections. Am. Soc. Microb. 2008:119–35. Washington D. C.
doi: 10.1128/9781555815561
Rubio D, Xu RH, Remakus S, Krouse TE, Truckenmiller ME, Thapa RJ, Balachandran S, Alcamí A, Norbury CC. L.J. Cross-talk between the type 1 interferon and nuclear factor Kappa B pathways confers resistance to a lethal virus infection. Cell Host Microbe. 2013;12(6):701–10. https://doi.org/10.1016/j.chom.2013.04.015
doi: 10.1016/j.chom.2013.04.015
Moschonas A, Ioannou M, Eliopoulos AG. CD40 stimulates a feed-forward NF-κB–driven molecular pathway that regulates IFN-β expression in carcinoma cells. J Immunol. 2012;188(11):5521–7. https://doi.org/10.4049/jimmunol.1200133
doi: 10.4049/jimmunol.1200133 pubmed: 22547704
Bertolusso R, Tian B, Zhao Y, Vergara L, Sabree A, Iwanaszko M, et al. Dynamic cross talk model of the epithelial innate immune response to double-stranded RNA stimulation: coordinated dynamics emerging from cell-level noise. PLoS ONE. 2014;9(4):e93396. https://doi.org/10.1371/journal.pone.0093396
doi: 10.1371/journal.pone.0093396 pubmed: 24710104 pmcid: 3977818
Gómez-Villamandos JC, Hervás J, Méndez A, Carrasco L, Villeda CJ, et al. Experimental African swine fever: apoptosis of lymphocytes and virus replication in other cells. J Gen Virol. 1995;76:2399–405.
doi: 10.1099/0022-1317-76-9-2399 pubmed: 7561784
Powell PP, Dixon LK, Parkhouse RM. An IkappaB homologue encoded by African swine fever virus provides a novel mechanism for downregulation of proinflamatory cytokine responses in host macrophages. J Virol. 1996;70:8527–33.
doi: 10.1128/jvi.70.12.8527-8533.1996 pubmed: 8970976 pmcid: 190944
Cabezón O, Muñoz-González S, Colom-Cadena A, Pérez-Simó M, Rosell R, Lavín S, Marco I, et al. African swine fever virus infection in classical swine fever subclinically infected wild boars. Vet Res. 2017;13:227. https://doi.org/10.1186/s12917-017-1150-0
doi: 10.1186/s12917-017-1150-0
Driessler F, Ventrom K, Sabat R, Asadullah K, Schottelius AJ. Molecular mechanisms of interleukin-10-mediated inhibition of NF-κB activity: a role for p50. Clin Exp Immunol. 2004;135:64–73.
doi: 10.1111/j.1365-2249.2004.02342.x pubmed: 14678266 pmcid: 1808913
McCleary S, Strong R, McCarthy RR, et al. Substitution of warthog NF-κB motifs into RELA of domestic pigs is not sufficient to confer resilience to African swine fever virus. Sci Rep. 2020;10:8951. https://doi.org/10.1038/s41598-020-658 .
doi: 10.1038/s41598-020-658 pubmed: 32488046 pmcid: 7265332

Auteurs

Patrick N Bisimwa (PN)

Molecular Biology Laboratory, Department of Animal Sciences and Production, Université Evangélique en Afrique, Bukavu, Democratic Republic of Congo. patrick.ntagereka@gmail.com.
Institut Supérieur de Dévelopement Rural de Kaziba, Kaziba, Democratic Republic of Congo. patrick.ntagereka@gmail.com.

Juliette R Ongus (JR)

Department of Medical Laboratory Sciences, Jomo Kenyatta University of Agriculture and Technology, Nairobi, Kenya.
Biotechnology Laboratory, Departement of Molecular Biology and Biotechnology, Pan African University Institute of Basic Sciences, Technology and Innovation, Nairobi, Kenya.

Ronald Tonui (R)

Department of Medical Laboratory Sciences, Jomo Kenyatta University of Agriculture and Technology, Nairobi, Kenya.

Espoir B Bisimwa (EB)

Molecular Biology Laboratory, Department of Animal Sciences and Production, Université Evangélique en Afrique, Bukavu, Democratic Republic of Congo.

Lucilla Steinaa (L)

Animal and Human Health Program, International Livestock Research Institute, Nairobi, Kenya.

Articles similaires

Genome, Chloroplast Phylogeny Genetic Markers Base Composition High-Throughput Nucleotide Sequencing
Robotic Surgical Procedures Animals Humans Telemedicine Models, Animal

Odour generalisation and detection dog training.

Lyn Caldicott, Thomas W Pike, Helen E Zulch et al.
1.00
Animals Odorants Dogs Generalization, Psychological Smell
Animals TOR Serine-Threonine Kinases Colorectal Neoplasms Colitis Mice

Classifications MeSH