Localization of conserved and variable antigenic domains of equine infectious anemia virus envelope glycoproteins using recombinant env-encoded protein fragments produced in Escherichia coli.
Abstract: Previous characterizations of equine infectious anemia virus (EIAV) glycoprotein variation by DNA sequence analysis and epitope mapping using monoclonal antibodies (MAbs) have revealed the presence of conserved and variable regions within the EIAV env gene. To extend these studies, fragments of the EIAV envelope proteins gp90 and gp45 were expressed in Escherichia coli and used in Western blot analysis with a diverse panel of equine immune sera to identify antigenic segments. All sera from EIAV-infected animals reacted with the carboxyl terminal portion of gp90 and the amino terminal portion of gp45, indicating the highly conserved and immunodominant nature of these regions. Other gp90 segments, both from conserved and variable env sequences, displayed variable reactivities with the panel of equine sera. A panel of MAbs was also used in Western blot assays with the recombinant protein fragments for physical localization of previously identified MAb epitopes. The binding sites of two neutralizing MAbs were localized to a highly variable region of gp90, while nonneutralizing epitopes were localized to conserved and variable regions of the envelope glycoproteins. These results, in addition to localizing important antigenic sites on EIAV glycoproteins, indicate that previously defined conserved and variable env nucleotide sequences indeed encode protein sequences constituting conserved and variable immunogens during persistent infection by EIAV.
Publication Date: 1989-10-01 PubMed ID: 2552661DOI: 10.1016/0042-6822(89)90203-1Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- Non-U.S. Gov't
- Research Support
- U.S. Gov't
- P.H.S.
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.
The research article discusses the identification and localization of the conserved and variable antigenic domains of the Equine Infectious Anemia Virus (EIAV) using fragments of the virus’s envelope proteins. With the help of recombinant proteins produced in Escherichia coli, the researchers were able to determine the reactivity and precise locations of these regions.
Overview of the Study
- The study aimed to characterize the variations in the EIAV glycoprotein by analyzing its DNA sequence and mapping it using monoclonal antibodies (MAbs) which led them to identify both conserved (unaltered) and variable regions within the EIAV env gene.
- The research aimed at expanding past studies by expressing fragments of the EIAV envelope proteins gp90 and gp45 within Escherichia coli.
- These expressed proteins were then used for Western blot analysis in combination with an array of equine immune sera to identify antigenic regions.
Findings of the Study
- All the sera from EIAV-infected animals reacted to the carboxyl terminal portion of gp90 and the amino terminal portion of gp45, showcasing the consistent and immunodominant nature of these regions.
- Different gp90 segments, originating from both conserved and variable env sequences, showed varied reactivity with equine sera.
- The binding sites of two neutralizing monoclonal antibodies were located within a highly variable region of gp90, whereas non-neutralizing epitopes were found in both conserved and variable areas of the envelope glycoproteins.
- This means that the regions that are defined as conservatively conserved and variable at the genetic level are also correspondingly variable or conserved at the level of immune responses.
Significance of the Research
- This research is fundamental in pointing out crucial antigenic sites on EIAV glycoproteins, providing a basis for potential future studies on EIAV vaccine development.
- The finding that the sequences categorized as conserved and variable reflect similar categorization at the protein immunogenic level means that any changes in these areas have direct implications for immunity, vaccination, and disease progression.
Cite This Article
APA
Payne SL, Rushlow K, Dhruva BR, Issel CJ, Montelaro RC.
(1989).
Localization of conserved and variable antigenic domains of equine infectious anemia virus envelope glycoproteins using recombinant env-encoded protein fragments produced in Escherichia coli.
Virology, 172(2), 609-615.
https://doi.org/10.1016/0042-6822(89)90203-1 Publication
Researcher Affiliations
- Department of Biochemistry, Louisiana State University, Baton Rouge.
MeSH Terms
- Animals
- Antibodies, Monoclonal / immunology
- Antigens, Viral / analysis
- Antigens, Viral / genetics
- Blotting, Western
- Cloning, Molecular
- Electrophoresis, Polyacrylamide Gel
- Escherichia coli / genetics
- Glycoproteins / genetics
- Glycoproteins / immunology
- Immune Sera / immunology
- Infectious Anemia Virus, Equine / genetics
- Infectious Anemia Virus, Equine / immunology
- Recombinant Fusion Proteins / genetics
- Recombinant Fusion Proteins / immunology
- Viral Envelope Proteins / genetics
- Viral Envelope Proteins / immunology
Grant Funding
- AI25850 / NIAID NIH HHS
- CA38851 / NCI NIH HHS
Citations
This article has been cited 22 times.- Craigo JK, Montelaro RC. Lessons in AIDS vaccine development learned from studies of equine infectious, anemia virus infection and immunity. Viruses 2013 Dec 2;5(12):2963-76.
- Craigo JK, Ezzelarab C, Montelaro RC. Development of a high throughput, semi-automated, infectious center cell-based ELISA for equine infectious anemia virus. J Virol Methods 2012 Nov;185(2):221-7.
- Cappelli K, Capomaccio S, Cook FR, Felicetti M, Marenzoni ML, Coppola G, Verini-Supplizi A, Coletti M, Passamonti F. Molecular detection, epidemiology, and genetic characterization of novel European field isolates of equine infectious anemia virus. J Clin Microbiol 2011 Jan;49(1):27-33.
- Craigo JK, Barnes S, Cook SJ, Issel CJ, Montelaro RC. Divergence, not diversity of an attenuated equine lentivirus vaccine strain correlates with protection from disease. Vaccine 2010 Nov 29;28(51):8095-104.
- Craigo JK, Barnes S, Zhang B, Cook SJ, Howe L, Issel CJ, Montelaro RC. An EIAV field isolate reveals much higher levels of subtype variability than currently reported for the equine lentivirus family. Retrovirology 2009 Oct 20;6:95.
- Brindley MA, Zhang B, Montelaro RC, Maury W. An equine infectious anemia virus variant superinfects cells through novel receptor interactions. J Virol 2008 Oct;82(19):9425-32.
- Payne SL, Pei XF, Jia B, Fagerness A, Fuller FJ. Influence of long terminal repeat and env on the virulence phenotype of equine infectious anemia virus. J Virol 2004 Mar;78(5):2478-85.
- Tencza SB, Islam KR, Kalia V, Nasir MS, Jolley ME, Montelaro RC. Development of a fluorescence polarization-based diagnostic assay for equine infectious anemia virus. J Clin Microbiol 2000 May;38(5):1854-9.
- 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.
- Zheng YH, Sentsui H, Nakaya T, Kono Y, Ikuta K. In vivo dynamics of equine infectious anemia viruses emerging during febrile episodes: insertions/duplications at the principal neutralizing domain. J Virol 1997 Jul;71(7):5031-9.
- 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.
- Carvalho M, Kirkland M, Derse D. Protein interactions with DNA elements in variant equine infectious anemia virus enhancers and their impact on transcriptional activity. J Virol 1993 Nov;67(11):6586-95.
- Sellon DC. Equine infectious anemia. Vet Clin North Am Equine Pract 1993 Aug;9(2):321-36.
- Sellon DC, Fuller FJ, McGuire TC. The immunopathogenesis of equine infectious anemia virus. Virus Res 1994 May;32(2):111-38.
- Lichtenstein DL, Rushlow KE, Cook RF, Raabe ML, Swardson CJ, Kociba GJ, Issel CJ, Montelaro RC. Replication in vitro and in vivo of an equine infectious anemia virus mutant deficient in dUTPase activity. J Virol 1995 May;69(5):2881-8.
- Chong YH, Ball JM, Issel CJ, Montelaro RC, Rushlow KE. Analysis of equine humoral immune responses to the transmembrane envelope glycoprotein (gp45) of equine infectious anemia virus. J Virol 1991 Feb;65(2):1013-8.
- Chong YH, Payne SL, Issel CJ, Montelaro RC, Rushlow KE. Characterization of the antigenic domains of the major core protein (p26) of equine infectious anemia virus. J Virol 1991 Feb;65(2):1007-12.
- Knowles DP Jr, Cheevers WP, McGuire TC, Brassfield AL, Harwood WG, Stem TA. Structure and genetic variability of envelope glycoproteins of two antigenic variants of caprine arthritis-encephalitis lentivirus. J Virol 1991 Nov;65(11):5744-50.
- Alexandersen S, Carpenter S. Characterization of variable regions in the envelope and S3 open reading frame of equine infectious anemia virus. J Virol 1991 Aug;65(8):4255-62.
- Ball JM, Rushlow KE, Issel CJ, Montelaro RC. Detailed mapping of the antigenicity of the surface unit glycoprotein of equine infectious anemia virus by using synthetic peptide strategies. J Virol 1992 Feb;66(2):732-42.
- Perry ST, Flaherty MT, Kelley MJ, Clabough DL, Tronick SR, Coggins L, Whetter L, Lengel CR, Fuller F. The surface envelope protein gene region of equine infectious anemia virus is not an important determinant of tropism in vitro. J Virol 1992 Jul;66(7):4085-97.
- McGuire TC, Knowles DP Jr, Davis WC, Brassfield AL, Stem TA, Cheevers WP. Transmembrane protein oligomers of caprine arthritis-encephalitis lentivirus are immunodominant in goats with progressive arthritis. J Virol 1992 May;66(5):3247-50.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists
