Characterization of an equine infectious anemia antigen extracted from infected horse spleen tissue.
Abstract: The spleens of horses infected with equine infectious anemia contain an antigen that is useful for a diagnostic immunodiffusion test. This antigen was extracted from the spleen by homogenization of the tissue, centrifugation, and precipitation from the supernatant fluid at 50% saturation with (NH(4))(2)SO(4). The antigen was purified by subjecting it to two cycles of electrophoresis in a continuous free-flow electrophoresis cell and finally filtering through a column of Sephadex G-200 gel. The antigen was found to be a small protein with a molecular weight of 27,500 and sedimentation coefficient of 2.1S. Its density was about 1.18 and its isoelectric point 5.8. At 45 to 50 C, it coagulated, losing its antigenicity. The antigen was useful for assaying antibody in the serum from infected horses by using the complement fixation test or the immunodiffusion test. Complement-fixing antibodies were found to be more transient than the precipitating antibodies.
Publication Date: 1971-11-01 PubMed ID: 5005308PubMed Central: PMC416346DOI: 10.1128/iai.4.5.528-531.1971Google Scholar: Lookup
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Summary
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The research paper provides a detailed analysis of an antigen found in the spleens of horses infected with equine infectious anemia, describing its extraction, purification, and characterization for use in diagnostic immunodiffusion tests.
Extraction of Antigen from Spleen
The researchers extracted an antigen from the spleen tissue of horses infected with equine infectious anemia by:
- Homogenizing the spleen tissue
- Centrifuging the homogenized tissue
- Using ammonium sulfate ((NH4)2SO4) to precipitate the antigen from the supernatant fluid at 50% saturation
Purification of Antigen
Post extraction, the antigen was purified through:
- Subjecting it to two cycles of electrophoresis in a continuous free-flow electrophoresis cell
- Filtering it through a column of Sephadex G-200 gel.
Characterization of the Antigen
The antigen was found to be a small protein with specific characteristics:
- It had a molecular weight of 27,500.
- Its sedimentation coefficient was 2.1S.
- Its density was about 1.18.
- Its isoelectric point was 5.8.
- At temperatures of 45 to 50 degrees Celsius, it coagulated and lost its antigenicity.
Use of the Antigen in Diagnostic Tests
The antigen was found to be useful for assaying antibodies in the serum from infected horses. The following tests were used:
- Complement fixation test
- Immunodiffusion test.
The research found complement-fixing antibodies to be more short-lived than the precipitating antibodies.
Cite This Article
APA
Norcross NL, Coggins L.
(1971).
Characterization of an equine infectious anemia antigen extracted from infected horse spleen tissue.
Infect Immun, 4(5), 528-531.
https://doi.org/10.1128/iai.4.5.528-531.1971 Publication
Researcher Affiliations
MeSH Terms
- Animals
- Antigens, Viral / analysis
- Antigens, Viral / isolation & purification
- Centrifugation, Density Gradient
- Complement Fixation Tests
- Electrophoresis
- Equine Infectious Anemia / immunology
- Filtration
- Horses / immunology
- Hot Temperature
- Immunodiffusion
- Isoelectric Focusing
- Molecular Weight
- Spleen / immunology
- Ultracentrifugation
References
This article includes 5 references
- Cowan KM, Graves JH. A third antigenic component associated with foot-and-mouth disease infection.. Virology 1966 Nov;30(3):528-40.
- Coggins L, Norcross NL. Immunodiffusion reaction in equine infectious anemia.. Cornell Vet 1970 Apr;60(2):330-5.
- Boulanger P, Bannister GL, Ruckerbauer GM, Corner AH. Equine infectious anemia: preliminary investigation of the complement-fixation test for the demonstration of antibodies and antigen.. Can J Comp Med 1969 Apr;33(2):148-54.
- Saxer E. [Current serologic aspects of infectious equine anemia in perissodactyla].. Schweiz Arch Tierheilkd 1966 Jun;108(6):331-4.
- Nakajima H, Ushimi C. Immunodiffusion studies of purified equine infectious anemia virus.. Infect Immun 1971 Mar;3(3):373-7.
Citations
This article has been cited 9 times.- Hu Z, Guo K, Du C, Sun J, Naletoski I, Chu X, Lin Y, Wang X, Barrandeguy M, Samuel M, Wang W, Lau PI, Wernery U, Raghavan R, Wang X. Development and evaluation of a blocking ELISA for serological diagnosis of equine infectious anemia. Appl Microbiol Biotechnol 2023 May;107(10):3305-3317.
- Nakajima H, Norcross NL, Coggins L. Demonstration of antigenic identity between purified equine infectious anemia virus and an antigen extracted from infected horse spleen. Infect Immun 1972 Sep;6(3):416-7.
- Malmquist WA, Barnett D, Becvar CS. Production of equine infectious anemia antigen in a persistently infected cell line. Arch Gesamte Virusforsch 1973;42(4):361-70.
- Nakajima H, Ushimi C, Fukunaga Y, Hirasawa K. Preparation of equine infectious anemia virus antigen for immunodiffusion test. Arch Gesamte Virusforsch 1973;42(4):339-45.
- Carrier SP, Bannister GL, Boulanger P. Equine infectious anemia: activity of liquid antigen extracts in the agar-gel immunodiffusion and complement-fixation tests. Can J Comp Med 1972 Oct;36(4):377-9.
- Sugiura T, Nakajima H. Purification of equine infectious anemia virus antigen by affinity chromatography. J Clin Microbiol 1977 Jun;5(6):635-9.
- Archer BG, Crawford TB, McGuire TC, Frazier ME. RNA-dependent DNA polymerase associated with equine infectious anemia virus. J Virol 1977 Apr;22(1):16-22.
- Charman HP, Bladen S, Gilden RV, Coggins L. Equine infectious anemia virus: evidence favoring classification as a retravirus. J Virol 1976 Sep;19(3):1073-9.
- Almaqhawi AA, El-Jalii IM, Al-Sabi MNS, Al-Ali A, Khalid AM, Abduljawad M, Shawaf T. Treatment evaluation using ultrasonographic scanning of the spleen in Arabian horses affected by babesiosis. Open Vet J 2025 Nov;15(11):5799-5805.
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