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Home / Equine Research Bank / Virus Genes / Amino acid mutations in the env gp90 protein that modify N-l...
Virus genes2016; 52(6); 814-822; doi: 10.1007/s11262-016-1382-2

Amino acid mutations in the env gp90 protein that modify N-linked glycosylation of the Chinese EIAV vaccine strain enhance resistance to neutralizing antibodies.

Abstract: The Chinese EIAV vaccine is an attenuated live virus vaccine obtained by serial passage of a virulent horse isolate (EIAV) in donkeys (EIAV) and, subsequently, in donkey cells in vitro. In this study, we compare the env gene of the original horse virulent virus (EIAV) with attenuated strains serially passaged in donkey MDM (EIAV) and donkey dermal cells (EIAV). Genetic comparisons among parental and attenuated strains found that vaccine strains contained amino acid substitutions/deletions in gp90 that resulted in a loss of three potential N-linked glycosylation sites, designated g5, g9, and g10. To investigate the biological significance of these changes, reverse-mutated viruses were constructed in the backbone of the EIAV infectious molecular clone (pLGFD3). The resulting virus stocks were characterized for replication efficiency in donkey dermal cells and donkey MDM, and were tested for sensitivity to neutralization using sera from two ponies experimentally infected with EIAV. Results clearly show that these mutations generated by site-directed mutagenesis resulted in cloned viruses with enhanced resistance to serum neutralizing antibodies that were also able to recognize parental viruses. This study indicates that these mutations played an important role in the attenuation of the EIAV vaccine strains.
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Publication Date: 2016-08-29 PubMed ID: 27572122DOI: 10.1007/s11262-016-1382-2Google Scholar: Lookup
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  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This study explores how certain amino acid changes in the envelope protein of the Chinese Equine Infectious Anemia Virus (EIAV) vaccine strain can enhance the strain’s resistance to neutralizing antibodies, thereby improving the vaccine’s effectiveness.

Context

  • This research focuses on the Chinese EIAV vaccine, a weakened live-virus inoculation created through repeated transmission of a virulent EIAV strain, sourced from horses, through donkeys and their cells.
  • The objective was to understand the genetic regulators of the EIAV vaccine’s resistance to neutralizing antibodies, which are part of the body’s immune response to fight off pathogens.

Study Design

  • The researchers compared the gene ‘env’ of the original horse-powered virus (EIAV) with the attenuated strains that were repeatedly passaged in two kinds of donkey cells (MDM and dermal cells).
  • This comparison sought to identify genetic variances between the virulent and attenuated strains, specifically looking for amino acid substitutions or deletions within the gp90 protein that could alter N-linked glycosylation – a process essential for protein folding and stability.

Findings

  • The research discovered that the vaccinated strains had amino acid changes in the gp90 protein that led to a loss of three potential N-linked glycosylation sites, termed g5, g9, and g10.
  • To comprehend the biological implications of these changes, the researchers built reverse-mutated viruses within the EIAV infectious molecular clone.
  • These genetically modified viruses were then evaluated for their replication efficiency within both donkey dermal cells and donkey MDM, and were tested for their resistance to neutralization using serum from two ponies infected with EIAV.

Conclusion

  • The observations reveal that the mutations made via site-directed mutagenesis resulted in cloned viruses that were more resistant to serum neutralizing antibodies and could detect the original viruses.
  • This indicates that these particular amino acid changes are significant contributors to the weakening, or attenuation, of the EIAV vaccine strains, implying their potentially crucial role in enhancing the vaccine’s efficacy.

Cite This Article

APA
Han X, Zhang P, Yu W, Xiang W, Li X. (2016). Amino acid mutations in the env gp90 protein that modify N-linked glycosylation of the Chinese EIAV vaccine strain enhance resistance to neutralizing antibodies. Virus Genes, 52(6), 814-822. https://doi.org/10.1007/s11262-016-1382-2

Publication

Virus genes
ISSN: 1572-994X
NlmUniqueID: 8803967
Country: United States
Language: English
Volume: 52
Issue: 6
Pages: 814-822

Researcher Affiliations

Han, Xiue
  • College of Food Science, Northeast Agricultural University, Harbin, 150030, China.
Zhang, Ping
  • College of Food Science, Northeast Agricultural University, Harbin, 150030, China.
Yu, Wei
  • College of Food Science, Northeast Agricultural University, Harbin, 150030, China.
Xiang, Wenhua
  • National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, 150030, China.
Li, Xiaodong
  • College of Food Science, Northeast Agricultural University, Harbin, 150030, China. hrblxd1@126.com.

MeSH Terms

  • Amino Acid Sequence
  • Amino Acid Substitution
  • Animals
  • Antibodies, Neutralizing / immunology
  • Antibodies, Viral / immunology
  • Cell Line
  • Cells, Cultured
  • Gene Products, env / chemistry
  • Gene Products, env / genetics
  • Gene Products, env / immunology
  • Gene Products, env / metabolism
  • Genome, Viral
  • Glycosylation
  • Horses
  • Infectious Anemia Virus, Equine / genetics
  • Infectious Anemia Virus, Equine / immunology
  • Mutation
  • Neutralization Tests
  • Viral Vaccines / genetics
  • Viral Vaccines / immunology

References

This article includes 29 references
  1. Wei X, Decker JM, Wang S, Hui H, Kappes JC, Wu X, Salazar-Gonzalez JF, Salazar MG, Kilby JM, Saag MS, Komarova NL, Nowak MA, Hahn BH, Kwong PD, Shaw GM. Antibody neutralization and escape by HIV-1.. Nature 2003 Mar 20;422(6929):307-12.
    pubmed: 12646921doi: 10.1038/nature01470google scholar: lookup
  2. Reitter JN, Desrosiers RC. Identification of replication-competent strains of simian immunodeficiency virus lacking multiple attachment sites for N-linked carbohydrates in variable regions 1 and 2 of the surface envelope protein.. J Virol 1998 Jul;72(7):5399-407.
    pubmed: 9620994doi: 10.1128/JVI.72.7.5399-5407.1998google scholar: lookup
  3. Cook RF, Leroux C, Cook SJ, Berger SL, Lichtenstein DL, Ghabrial NN, Montelaro RC, Issel CJ. Development and characterization of an in vivo pathogenic molecular clone of equine infectious anemia virus.. J Virol 1998 Feb;72(2):1383-93.
    pubmed: 9445039doi: 10.1128/JVI.72.2.1383-1393.1998google scholar: lookup
  4. Wei L, Fan X, Lu X, Zhao L, Xiang W, Zhang X, Xue F, Shao Y, Shen R, Wang X. Genetic variation in the long terminal repeat associated with the transition of Chinese equine infectious anemia virus from virulence to avirulence.. Virus Genes 2009 Apr;38(2):285-8.
    pubmed: 19130201doi: 10.1007/s11262-008-0317-ygoogle scholar: lookup
  5. Howe L, Leroux C, Issel CJ, Montelaro RC. Equine infectious anemia virus envelope evolution in vivo during persistent infection progressively increases resistance to in vitro serum antibody neutralization as a dominant phenotype.. J Virol 2002 Nov;76(21):10588-97.
    pubmed: 12368301doi: 10.1128/jvi.76.21.10588-10597.2002google scholar: lookup
  6. Leroux C, Issel CJ, Montelaro RC. Novel and dynamic evolution of equine infectious anemia virus genomic quasispecies associated with sequential disease cycles in an experimentally infected pony.. J Virol 1997 Dec;71(12):9627-39.
    pubmed: 9371627doi: 10.1128/JVI.71.12.9627-9639.1997google scholar: lookup
  7. Ma J, Jiang C, Lin Y, Wang X, Zhao L, Xiang W, Shao Y, Shen R, Kong X, Zhou J. In vivo evolution of the gp90 gene and consistently low plasma viral load during transient immune suppression demonstrate the safety of an attenuated equine infectious anemia virus (EIAV) vaccine.. Arch Virol 2009;154(5):867-73.
    pubmed: 19363668doi: 10.1007/s00705-009-0378-9google scholar: lookup
  8. Craigo JK, Leroux C, Howe L, Steckbeck JD, Cook SJ, Issel CJ, Montelaro RC. Transient immune suppression of inapparent carriers infected with a principal neutralizing domain-deficient equine infectious anaemia virus induces neutralizing antibodies and lowers steady-state virus replication.. J Gen Virol 2002 Jun;83(Pt 6):1353-1359.
    pubmed: 12029150doi: 10.1099/0022-1317-83-6-1353google scholar: lookup
  9. Abel K, Compton L, Rourke T, Montefiori D, Lu D, Rothaeusler K, Fritts L, Bost K, Miller CJ. Simian-human immunodeficiency virus SHIV89.6-induced protection against intravaginal challenge with pathogenic SIVmac239 is independent of the route of immunization and is associated with a combination of cytotoxic T-lymphocyte and alpha interferon responses.. J Virol 2003 Mar;77(5):3099-118.
    pubmed: 12584336doi: 10.1128/jvi.77.5.3099-3118.2003google scholar: lookup
  10. Wang X, Wang S, Lin Y, Jiang C, Ma J, Zhao L, Lv X, Wang F, Shen R, Kong X, Zhou J. Genomic comparison between attenuated Chinese equine infectious anemia virus vaccine strains and their parental virulent strains.. Arch Virol 2011 Feb;156(2):353-7.
    pubmed: 21136127doi: 10.1007/s00705-010-0877-8google scholar: lookup
  11. Harrold SM, Cook SJ, Cook RF, Rushlow KE, Issel CJ, Montelaro RC. Tissue sites of persistent infection and active replication of equine infectious anemia virus during acute disease and asymptomatic infection in experimentally infected equids.. J Virol 2000 Apr;74(7):3112-21.
    pubmed: 10708426doi: 10.1128/jvi.74.7.3112-3121.2000google scholar: lookup
  12. Cook RF, Cook SJ, Berger SL, Leroux C, Ghabrial NN, Gantz M, Bolin PS, Mousel MR, Montelaro RC, Issel CJ. Enhancement of equine infectious anemia virus virulence by identification and removal of suboptimal nucleotides.. Virology 2003 Sep 1;313(2):588-603.
    pubmed: 12954224doi: 10.1016/s0042-6822(03)00351-9google scholar: lookup
  13. Wang X, Wang S, Lin Y, Jiang C, Ma J, Zhao L, Lv X, Wang F, Shen R, Zhou J. Unique evolution characteristics of the envelope protein of EIAV(LN₄₀), a virulent strain of equine infectious anemia virus.. Virus Genes 2011 Apr;42(2):220-8.
    pubmed: 21369830doi: 10.1007/s11262-010-0563-7google scholar: lookup
  14. Johnson WE, Sanford H, Schwall L, Burton DR, Parren PW, Robinson JE, Desrosiers RC. Assorted mutations in the envelope gene of simian immunodeficiency virus lead to loss of neutralization resistance against antibodies representing a broad spectrum of specificities.. J Virol 2003 Sep;77(18):9993-10003.
    pubmed: 12941910doi: 10.1128/jvi.77.18.9993-10003.2003google scholar: lookup
  15. Spyrou V, Papanastassopoulou M, Koumbati M, Nikolakaki SV, Koptopoulos G. Molecular analysis of the proviral DNA of equine infectious anemia virus in mules in Greece.. Virus Res 2005 Jan;107(1):63-72.
    pubmed: 15567035doi: 10.1016/j.virusres.2004.06.016google scholar: lookup
  16. Wang XF, Jiang CG, Guo W, Xiang W, Lv XL, Zhao LP, Wang FL, Kong XG, Zhang XY, Shao YM, Zhou JH. [Comparison of proviral genomes between the Chinese EIAV donkey leukocyte-attenuated vaccine and its parental virulent strain].. Bing Du Xue Bao 2008 Nov;24(6):443-50.
    pubmed: 19226953
  17. Wang XF, Lin YZ, Li Q, Liu Q, Zhao WW, Du C, Chen J, Wang X, Zhou JH. Genetic Evolution during the development of an attenuated EIAV vaccine.. Retrovirology 2016 Feb 3;13:9.
    pubmed: 26842878doi: 10.1186/s12977-016-0240-6google scholar: lookup
  18. Craigo JK, Zhang B, Barnes S, Tagmyer TL, Cook SJ, Issel CJ, Montelaro RC. Envelope variation as a primary determinant of lentiviral vaccine efficacy.. Proc Natl Acad Sci U S A 2007 Sep 18;104(38):15105-10.
    pubmed: 17846425doi: 10.1073/pnas.0706449104google scholar: lookup
  19. Ma J, Shi N, Jiang CG, Lin YZ, Wang XF, Wang S, Lv XL, Zhao LP, Shao YM, Kong XG, Zhou JH, Shen RX. A proviral derivative from a reference attenuated EIAV vaccine strain failed to elicit protective immunity.. Virology 2011 Feb 5;410(1):96-106.
    pubmed: 21094511doi: 10.1016/j.virol.2010.10.032google scholar: lookup
  20. Hammond SA, Li F, McKeon BM Sr, Cook SJ, Issel CJ, Montelaro RC. Immune responses and viral replication in long-term inapparent carrier ponies inoculated with equine infectious anemia virus.. J Virol 2000 Jul;74(13):5968-81.
    pubmed: 10846078doi: 10.1128/jvi.74.13.5968-5981.2000google scholar: lookup
  21. Leroux C, Craigo JK, Issel CJ, Montelaro RC. Equine infectious anemia virus genomic evolution in progressor and nonprogressor ponies.. J Virol 2001 May;75(10):4570-83.
    pubmed: 11312327doi: 10.1128/JVI.75.10.4570-4583.2001google scholar: lookup
  22. Hussain KA, Issel CJ, Schnorr KL, Rwambo PM, Montelaro RC. Antigenic analysis of equine infectious anemia virus (EIAV) variants by using monoclonal antibodies: epitopes of glycoprotein gp90 of EIAV stimulate neutralizing antibodies.. J Virol 1987 Oct;61(10):2956-61.
    pubmed: 2442410doi: 10.1128/JVI.61.10.2956-2961.1987google scholar: lookup
  23. Lichtenstein DL, Issel CJ, Montelaro RC. Genomic quasispecies associated with the initiation of infection and disease in ponies experimentally infected with equine infectious anemia virus.. J Virol 1996 Jun;70(6):3346-54.
    pubmed: 8648664doi: 10.1128/JVI.70.6.3346-3354.1996google scholar: lookup
  24. Pikora CA. Glycosylation of the ENV spike of primate immunodeficiency viruses and antibody neutralization.. Curr HIV Res 2004 Jul;2(3):243-54.
    pubmed: 15279588doi: 10.2174/1570162043351264google scholar: lookup
  25. Zheng YH, Sentsui H, Kono Y, Ikuta K. Mutations occurring during serial passage of Japanese equine infectious anemia virus in primary horse macrophages.. Virus Res 2000 Jun;68(1):93-8.
    pubmed: 10930666doi: 10.1016/s0168-1702(00)00147-7google scholar: lookup
  26. Liang H, He X, Shen RX, Shen T, Tong X, Ma Y, Xiang WH, Zhang XY, Shao YM. Combined amino acid mutations occurring in the envelope closely correlate with pathogenicity of EIAV.. Arch Virol 2006 Jul;151(7):1387-403.
    pubmed: 16502285doi: 10.1007/s00705-005-0718-3google scholar: lookup
  27. Gao X, Jiang CG, Han XE, Zhao LP, Shen RX, Xiang WH, Zhou JH. [Construction and in vitro evaluation of an infectious clone of the equine infectious anemia virus vaccine strain EIAV(FDDV) with four reverse-mutated vaccine-specific sites in the S2 gene].. Bing Du Xue Bao 2009 Jul;25(4):309-15.
    pubmed: 19769166
  28. Qi X, Wang X, Wang S, Lin Y, Jiang C, Ma J, Zhao L, Lv X, Shen R, Wang F, Kong X, Su Z, Zhou J. Genomic analysis of an effective lentiviral vaccine-attenuated equine infectious anemia virus vaccine EIAV FDDV13.. Virus Genes 2010 Aug;41(1):86-98.
    pubmed: 20526660doi: 10.1007/s11262-010-0491-6google scholar: lookup
  29. Rwambo PM, Issel CJ, Hussain KA, Montelaro RC. In vitro isolation of a neutralization escape mutant of equine infectious anemia virus (EIAV).. Arch Virol 1990;111(3-4):275-80.
    pubmed: 1693846doi: 10.1007/BF01311062google scholar: lookup

Citations

This article has been cited 1 times.
  1. Liu C, Wang XF, Wang Y, Chen J, Zhong Z, Lin Y, Wang X. Characterization of EIAV env Quasispecies during Long-Term Passage In Vitro: Gradual Loss of Pathogenicity.. Viruses 2019 Apr 24;11(4).
    doi: 10.3390/v11040380pubmed: 31022927google scholar: lookup
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