Purification and antigenicity of an M-like protein of Streptococcus equi.
Abstract: A cell wall component of Streptococcus equi analogous to the M protein of group A streptococci has been identified and purified. A highly purified product has been obtained from cells by hot acid extraction, followed by acid precipitation, ammonium sulfate fractionation, and column chromatography. This product reacts with S. equi antiserum. The existence of this fraction in S. equi has been confirmed by the failure of trypsin-treated cells and their extracts to remove the long-chaining capacity of S. equi antiserum. The antigenicity of this M-like protein when incorporated in adjuvant has been assessed in rabbits and horses. In the rabbit, multiple doses of as low as 10 mug resulted in the production of serum capable of inducing long-chain formation and opsonizing and mouse protective antibody. Two doses of 100 mug elicited similar responses. The bactericidal capacity of this serum could be eliminated by absorption with whole S. equi cells or their extracts, but not by absorption with trypsin-treated cells or extracts therefrom. Horses vaccinated 15 months previously with a whole-cell killed vaccine and given 200 mug of the purified protein intramuscularly showed evidence of an anamnestic response to the antigen.
Publication Date: 1974-07-01 PubMed ID: 4210335PubMed Central: PMC414966DOI: 10.1128/iai.10.1.116-122.1974Google Scholar: Lookup
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- Journal Article
Summary
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This study investigates the identification, purification and antigenic potential of an M-like protein found in the cell wall of the Streptococcus equi bacterium, compared to the M protein of group A streptococci. The antigenic response to this protein was evaluated in rabbits and horses.
Identification and Purification of the M-like Protein
- The researchers discovered a component in the cell wall of Streptococcus equi, which was similar to the M protein of group A streptococci.
- To isolate this protein, the scientists used methods such as hot acid extraction, acid precipitation, ammonium sulfate fractionation, and column chromatography.
- The purified product reacted with S. equi antiserum, confirming the existence of the protein in the bacterium.
- Further evidence of the protein’s existence was provided by tests with trypsin-treated cells and their extracts which did not remove the long-chaining capacity of S. equi antiserum, another characteristic of M proteins.
Assessment of Antigenicity
- The researchers studied the antigenicity or immunogenic potential of the M-like protein by evaluating responses in rabbits and horses.
- When small doses, as low as 10 micrograms, of the protein were introduced in rabbits multiple times, it resulted in the production of serum that induced long-chain formation, opsonizing, and mouse protective antibodies.
- Horses that were previously vaccinated with a killed-whole cell S. equi vaccine showed an anamnestic or memory immune response when given a 200 microgram dose of the purified protein.
- Similar immunogenic responses were observed in animals given two doses of 100 micrograms.
- The bactericidal capacity of the serum in rabbits could be eliminated by absorption with S. equi cells or their extracts, indicating the specific targeting action of the protein.
Significant Findings and Future Research
- The study contributes significant insights into the antigenic properties of the M-like protein found in the S. equi bacterium. The results could potentially advance the development of new vaccine strategies against the bacterium that causes strangles, a severe respiratory infection in horses.
- Further research might explore the implications of the anamnestic response observed in horses, with potential for enhancing vaccine efficacy and for understanding the disease pathogenesis.
Cite This Article
APA
Woolcock JB.
(1974).
Purification and antigenicity of an M-like protein of Streptococcus equi.
Infect Immun, 10(1), 116-122.
https://doi.org/10.1128/iai.10.1.116-122.1974 Publication
Researcher Affiliations
MeSH Terms
- Absorption
- Animals
- Antibodies, Bacterial / analysis
- Bacterial Vaccines / isolation & purification
- Bacteriological Techniques
- Blood Bactericidal Activity
- Cell Wall / analysis
- Chromatography
- Ethanol
- Horses
- Immune Sera / pharmacology
- Immunity, Maternally-Acquired
- Immunization Schedule
- Immunodiffusion
- Immunologic Techniques
- Methylcellulose
- Rabbits
- Streptococcus / metabolism
- Tissue Extracts
- Viral Proteins / isolation & purification
References
This article includes 10 references
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Citations
This article has been cited 13 times.- Baker BS, Brown DW, Fischetti VA, Ovigne JM, Porter W, Powles A, Fry L. Skin T cell proliferative response to M protein and other cell wall and membrane proteins of group A streptococci in chronic plaque psoriasis. Clin Exp Immunol 2001 Jun;124(3):516-21.
- Boschwitz JS, Timoney JF. Inhibition of C3 deposition on Streptococcus equi subsp. equi by M protein: a mechanism for survival in equine blood. Infect Immun 1994 Aug;62(8):3515-20.
- Srivastava SK, Barnum DA. The serological response of foals to vaccination against strangles. Can J Comp Med 1981 Jan;45(1):20-5.
- Srivastava SK, Barnum DA. Lymphocyte stimulation response in horses against phytohaemagglutinin and M protein of Streptococcus equi using whole blood. Can J Comp Med 1982 Jan;46(1):51-6.
- Srivastava SK, Barnum DA. Studies on the immunogenicity of Streptococcus equi vaccines in foals. Can J Comp Med 1985 Oct;49(4):351-6.
- Timoney JF, Trachman J. Immunologically reactive proteins of Streptococcus equi. Infect Immun 1985 Apr;48(1):29-34.
- Scott JR, Pulliam WM, Hollingshead SK, Fischetti VA. Relationship of M protein genes in group A streptococci. Proc Natl Acad Sci U S A 1985 Mar;82(6):1822-6.
- Jones KF, Fischetti VA. Biological and immunochemical identity of M protein on group G streptococci with M protein on group A streptococci. Infect Immun 1987 Mar;55(3):502-6.
- Galán JE, Timoney JF. Immunologic and genetic comparison of Streptococcus equi isolates from the United States and Europe. J Clin Microbiol 1988 Jun;26(6):1142-6.
- Galán JE, Timoney JF. Molecular analysis of the M protein of Streptococcus equi and cloning and expression of the M protein gene in Escherichia coli. Infect Immun 1987 Dec;55(12):3181-7.
- Fischetti VA. Streptococcal M protein: molecular design and biological behavior. Clin Microbiol Rev 1989 Jul;2(3):285-314.
- Woods RD, Ross RF. Purification and serological characterization of a type-specific antigen of Streptococcus equisimilis. Infect Immun 1975 Oct;12(4):881-7.
- Soliman R, Yousef M, Gelil SA, Aboul-Ella H. Development of novel Streptococcus equi vaccines with an assessment of their immunizing potentials and protective efficacies. BMC Vet Res 2024 May 3;20(1):173.
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