Identification of 15- to 17-kilodalton antigens associated with virulent Rhodococcus equi.
Abstract: Antigens of Rhodococcus equi were analyzed by immunoblotting with naturally infected foal sera. Immunoblots of whole-cell antigen preparations of clinical isolates of R. equi revealed that major protein bands with molecular masses of 15 to 17 kDa were present in all clinical isolates tested and all isolates virulent for mice. In contrast, the 15- to 17-kDa antigens were not identified by immunoblotting in ATCC 6939, a type strain of R. equi that was avirulent for mice. Whole-cell antigens of 102 environmental isolates were investigated by immunoblotting and the mouse pathogenicity test. Twenty-five of these isolates were demonstrated to contain the 15- to 17-kDa antigens by immunoblotting and were virulent for mice. The remaining 77 environmental isolates lacked the 15- to 17-kDa antigens and were avirulent for mice. These data suggest that the diffuse 15- to 17-kDa proteins are virulence-associated antigens with immunogenicity in foals and that they may be useful in marking virulent R. equi contamination in the environment of a horse-breeding farm.
Publication Date: 1991-03-01 PubMed ID: 2037660PubMed Central: PMC269796DOI: 10.1128/jcm.29.3.439-443.1991Google 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
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 is about the identification of particular antigens in the Rhodococcus equi bacterium that are associated with its virulence or disease causing capability. The presence of these 15 to 17 kDa proteins distinguishes virulent forms of R. equi from non-virulent strains, and they could serve as markers for the presence of this harmful bacterium in the horse-breeding farm environment.
Analysis of R. equi Antigens
- The research began with the study of antigens present in the Rhodococcus equi bacterium by utilizing immunoblotting, a technique that allows identification of proteins in a mixture. Serum samples from naturally infected foals were used for this blotting process.
- Immunoblots of whole-cell proteins extracted from clinical isolates of R. equi highlighted that there were major protein bands of 15 to 17 kDa size in all clinical isolates tested. These protein bands were also present in all isolates that were harmfully virulent for mice.
Contrasting Virulent and Avirulent Strains
- On the other hand, a comparison was made with ATCC 6939 which is a known type strain of R. equi but does not cause infection in mice. This strain did not show the presence of 15 to 17 kDa antigens in immunoblotting.
- This comparison signifies that these proteins are particularly linked to the virulence of this bacterium, thus differentiating disease-causing versions from harmless ones.
Studying Environmental Isolates
- Moving beyond clinical isolates, the researchers directed their focus towards environmental isolates. They studied whole-cell antigens of 102 environmental isolates using the same technique of immunoblotting and carried out testing for pathogenicity in mice.
- Out of these 102, 25 isolates were carrying the 15 to 17 kDa antigens as per the immunoblotting results and were also determined to be harmful for mice. However, the rest of the 77 isolates didn’t have the 15 to 17 kDa antigens and were found to be harmless for mice.
Significance of the Research
- The consistent finding of 15 to 17 kDa proteins in virulent clinical and environmental samples of R. equi suggests that these proteins hold substantial role in causing infection.
- The proteins also exhibited immunogenicity in foals, triggering an immune response. As such, these antigens can potentially serve as markers for the virulent strains of R. equi, aiding in the identification and control of this bacterium in the environment of horse breeding farms.
Cite This Article
APA
Takai S, Koike K, Ohbushi S, Izumi C, Tsubaki S.
(1991).
Identification of 15- to 17-kilodalton antigens associated with virulent Rhodococcus equi.
J Clin Microbiol, 29(3), 439-443.
https://doi.org/10.1128/jcm.29.3.439-443.1991 Publication
Researcher Affiliations
- Department of Animal Hygiene, School of Veterinary Medicine and Animal Sciences, Kitasato University, Aomori, Japan.
MeSH Terms
- Actinomycetales Infections / immunology
- Actinomycetales Infections / veterinary
- Animals
- Antigens, Bacterial / chemistry
- Antigens, Bacterial / isolation & purification
- Biomarkers
- Horse Diseases / immunology
- Horses
- Immunoblotting
- Lung / immunology
- Lymph Nodes / immunology
- Mice
- Molecular Weight
- Rhodococcus / immunology
- Rhodococcus / isolation & purification
- Rhodococcus / pathogenicity
- Serotyping
- Soil Microbiology
- Virulence / immunology
References
This article includes 22 references
- Takai S, Michizoe T, Matsumura K, Nagai M, Sato H, Tsubaki S. Correlation of in vitro properties of Rhodococcus (Corynebacterium) equi with virulence for mice.. Microbiol Immunol 1985;29(12):1175-84.
- Martens RJ, Martens JG, Fiske RA, Hietala SK. Rhodococcus equi foal pneumonia: protective effects of immune plasma in experimentally infected foals.. Equine Vet J 1989 Jul;21(4):249-55.
- Takai S, Narita K, Ando K, Tsubaki S. Ecology of Rhodococcus (Corynebacterium) equi in soil on a horse-breeding farm.. Vet Microbiol 1986 Jul;12(2):169-77.
- Chirino-Trejo JM, Prescott JF. Polyacrylamide gel electrophoresis of whole-cell preparations of Rhodococcus equi.. Can J Vet Res 1987 Jul;51(3):297-300.
- Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.. Nature 1970 Aug 15;227(5259):680-5.
- Yanagawa R, Honda E. Presence of pili in species of human and animal parasites and pathogens of the genuscorynebacterium.. Infect Immun 1976 Apr;13(4):1293-5.
- Woolcock JB, Mutimer MD. The capsules of Corynebacterium equi and Streptococcus equi.. J Gen Microbiol 1978 Nov;109(1):127-30.
- Bernheimer AW, Linder R, Avigad LS. Stepwise degradation of membrane sphingomyelin by corynebacterial phospholipases.. Infect Immun 1980 Jul;29(1):123-31.
- Prescott JF. Capsular serotypes of Corynebacterium equi.. Can J Comp Med 1981 Apr;45(2):130-4.
- Martens RJ, Fiske RA, Renshaw HW. Experimental subacute foal pneumonia induced by aerosol administration of Corynebacterium equi.. Equine Vet J 1982 Apr;14(2):111-6.
- Prescott JF, Lastra M, Barksdale L. Equi factors in the identification of Corynebacterium equi Magnusson.. J Clin Microbiol 1982 Nov;16(5):988-90.
- Nakazawa M, Haritani M, Sugimoto C, Isayama Y. Virulence of Rhodococcus equi for mice.. Nihon Juigaku Zasshi 1983 Oct;45(5):679-82.
- Nakazawa M, Kubo M, Sugimoto C, Isayama Y. Serogrouping of Rhodococcus equi.. Microbiol Immunol 1983;27(10):837-46.
- Prescott JF, Coshan-Gauthier R, Barksdale L. Antibody to equi factor(s) in the diagnosis of Corynebacterium equi pneumonia of foals.. Can J Comp Med 1984 Oct;48(4):370-3.
- Ellenberger MA, Kaeberle ML, Roth JA. Equine humoral immune response to Rhodococcus (Corynebacterium) equi.. Am J Vet Res 1984 Nov;45(11):2428-30.
- Hietala SK, Ardans AA, Sansome A. Detection of Corynebacterium equi-specific antibody in horses by enzyme-linked immunosorbent assay.. Am J Vet Res 1985 Jan;46(1):13-5.
- Skalka B, Svastová A. Serodiagnostics of Corynebacterium (Rhodococcus) equi.. Zentralbl Veterinarmed B 1985 Feb;32(2):137-42.
- Takai S, Kawazu S, Tsubaki S. Enzyme-linked immunosorbent assay for diagnosis of Corynebacterium (Rhodococcus) equi infection in foals.. Am J Vet Res 1985 Oct;46(10):2166-70.
- Chirino-Trejo JM, Prescott JF. Antibody response of horses to Rhodococcus equi antigens.. Can J Vet Res 1987 Jul;51(3):301-5.
- Bowles PM, Woolcock JB, Mutimer MD. Experimental infection of mice with Rhodococcus equi: differences in virulence between strains.. Vet Microbiol 1987 Aug;14(3):259-68.
- Takai S, Kawazu S, Tsubaki S. Humoral immune response of foals to experimental infection with Rhodococcus equi.. Vet Microbiol 1987 Aug;14(3):321-7.
- Takai S, Kawazu S, Tsubaki S. Immunoglobulin and specific antibody responses to Rhodococcus (Corynebacterium) equi infection in foals as measured by enzyme-linked immunosorbent assay.. J Clin Microbiol 1986 May;23(5):943-7.
Citations
This article has been cited 47 times.- Salazar-Rodríguez D, Aleaga-Santiesteban Y, Iglesias E, Plascencia-Hernández A, Pérez-Gómez HR, Calderón EJ, Vázquez-Boland JA, de Armas Y. Virulence Plasmids of Rhodococcus equi Isolates From Cuban Patients With AIDS.. Front Vet Sci 2021;8:628239.
- Bujold AR, Lani NR, Sanz MG. Strain-to-strain variation of Rhodococcus equi growth and biofilm formation in vitro.. BMC Res Notes 2019 Aug 19;12(1):519.
- Willingham-Lane JM, Coulson GB, Hondalus MK. Identification of a VapA virulence factor functional homolog in Rhodococcus equi isolates housing the pVAPB plasmid.. PLoS One 2018;13(10):e0204475.
- Kalinowski M, Grądzki Z, Jarosz Ł, Adaszek Ł. Molecular analysis of the chromosomal 16S rRNA gene and vapA plasmid gene of Polish field strains of R. equi.. PLoS One 2018;13(9):e0204024.
- Bryan LK, Alexander ER, Lawhon SD, Cohen ND. Detection of vapN in Rhodococcus equi isolates cultured from humans.. PLoS One 2018;13(1):e0190829.
- Wright LM, Carpinone EM, Bennett TL, Hondalus MK, Starai VJ. VapA of Rhodococcus equi binds phosphatidic acid.. Mol Microbiol 2018 Feb;107(3):428-444.
- Willingham-Lane JM, Berghaus LJ, Giguère S, Hondalus MK. Influence of Plasmid Type on the Replication of Rhodococcus equi in Host Macrophages.. mSphere 2016 Sep-Oct;1(5).
- Giles C, Ndi O, Barton MD, Vanniasinkam T. An Adenoviral Vector Based Vaccine for Rhodococcus equi.. PLoS One 2016;11(3):e0152149.
- Okoko T, Blagova EV, Whittingham JL, Dover LG, Wilkinson AJ. Structural characterisation of the virulence-associated protein VapG from the horse pathogen Rhodococcus equi.. Vet Microbiol 2015 Aug 31;179(1-2):42-52.
- Van Tyne D, Gilmore MS. Virulence Plasmids of Nonsporulating Gram-Positive Pathogens.. Microbiol Spectr 2014 Oct;2(5).
- Whittingham JL, Blagova EV, Finn CE, Luo H, Miranda-CasoLuengo R, Turkenburg JP, Leech AP, Walton PH, Murzin AG, Meijer WG, Wilkinson AJ. Structure of the virulence-associated protein VapD from the intracellular pathogen Rhodococcus equi.. Acta Crystallogr D Biol Crystallogr 2014 Aug;70(Pt 8):2139-51.
- Sydor T, von Bargen K, Hsu FF, Huth G, Holst O, Wohlmann J, Becken U, Dykstra T, Söhl K, Lindner B, Prescott JF, Schaible UE, Utermöhlen O, Haas A. Diversion of phagosome trafficking by pathogenic Rhodococcus equi depends on mycolic acid chain length.. Cell Microbiol 2013 Mar;15(3):458-73.
- Kuskie KR, Smith JL, Wang N, Carter CN, Chaffin MK, Slovis NM, Stepusin RS, Cattoi AE, Takai S, Cohen ND. Effects of location for collection of air samples on a farm and time of day of sample collection on airborne concentrations of virulent Rhodococcus equi at two horse breeding farms.. Am J Vet Res 2011 Jan;72(1):73-9.
- Poolkhet C, Chumsing S, Wajjwalku W, Minato C, Otsu Y, Takai S. Plasmid Profiles and Prevalence of Intermediately Virulent Rhodococcus equi from Pigs in Nakhonpathom Province, Thailand: Identification of a New Variant of the 70-kb Virulence Plasmid, Type 18.. Vet Med Int 2010;2010:491624.
- Coulson GB, Agarwal S, Hondalus MK. Characterization of the role of the pathogenicity island and vapG in the virulence of the intracellular actinomycete pathogen Rhodococcus equi.. Infect Immun 2010 Aug;78(8):3323-34.
- Hong Y, Hondalus MK. Site-specific integration of Streptomyces PhiC31 integrase-based vectors in the chromosome of Rhodococcus equi.. FEMS Microbiol Lett 2008 Oct;287(1):63-8.
- Cotter PD, Hill C. Surviving the acid test: responses of gram-positive bacteria to low pH.. Microbiol Mol Biol Rev 2003 Sep;67(3):429-53, table of contents.
- Hooper-McGrevy KE, Wilkie BN, Prescott JF. Immunoglobulin G subisotype responses of pneumonic and healthy, exposed foals and adult horses to Rhodococcus equi virulence-associated proteins.. Clin Diagn Lab Immunol 2003 May;10(3):345-51.
- Lopez AM, Hines MT, Palmer GH, Alperin DC, Hines SA. Identification of pulmonary T-lymphocyte and serum antibody isotype responses associated with protection against Rhodococcus equi.. Clin Diagn Lab Immunol 2002 Nov;9(6):1270-6.
- Morton AC, Begg AP, Anderson GA, Takai S, Lämmler C, Browning GF. Epidemiology of Rhodococcus equi strains on Thoroughbred horse farms.. Appl Environ Microbiol 2001 May;67(5):2167-75.
- Vanniasinkam T, Barton MD, Heuzenroeder MW. B-Cell epitope mapping of the VapA protein of Rhodococcus equi: implications for early detection of R. equi disease in foals.. J Clin Microbiol 2001 Apr;39(4):1633-7.
- Byrne BA, Prescott JF, Palmer GH, Takai S, Nicholson VM, Alperin DC, Hines SA. Virulence plasmid of Rhodococcus equi contains inducible gene family encoding secreted proteins.. Infect Immun 2001 Feb;69(2):650-6.
- Takai S, Hines SA, Sekizaki T, Nicholson VM, Alperin DA, Osaki M, Takamatsu D, Nakamura M, Suzuki K, Ogino N, Kakuda T, Dan H, Prescott JF. DNA sequence and comparison of virulence plasmids from Rhodococcus equi ATCC 33701 and 103.. Infect Immun 2000 Dec;68(12):6840-7.
- Takai S, Shoda M, Sasaki Y, Tsubaki S, Fortier G, Pronost S, Rahal K, Becu T, Begg A, Browning G, Nicholson VM, Prescott JF. Restriction fragment length polymorphisms of virulence plasmids in Rhodococcus equi.. J Clin Microbiol 1999 Oct;37(10):3417-20.
- Giguère S, Hondalus MK, Yager JA, Darrah P, Mosser DM, Prescott JF. Role of the 85-kilobase plasmid and plasmid-encoded virulence-associated protein A in intracellular survival and virulence of Rhodococcus equi.. Infect Immun 1999 Jul;67(7):3548-57.
- Kasuga-Aoki H, Takai S, Sasaki Y, Tsubaki S, Madarame H, Nakane A. Tumour necrosis factor and interferon-gamma are required in host resistance against virulent Rhodococcus equi infection in mice: cytokine production depends on the virulence levels of R. equi.. Immunology 1999 Jan;96(1):122-7.
- Takai S, Takeda K, Nakano Y, Karasawa T, Furugoori J, Sasaki Y, Tsubaki S, Higuchi T, Anzai T, Wada R, Kamada M. Emergence of rifampin-resistant Rhodococcus equi in an infected foal.. J Clin Microbiol 1997 Jul;35(7):1904-8.
- Haites RE, Muscatello G, Begg AP, Browning GF. Prevalence of the virulence-associated gene of Rhodococcus equi in isolates from infected foals.. J Clin Microbiol 1997 Jun;35(6):1642-4.
- Vullo V, Mastroianni CM, Lichtner M, Mengoni F, Chiappini E, D'Agostino C, Delia S. Serologic responses to Rhodococcus equi in individuals with and without human immunodeficiency virus infection.. Eur J Clin Microbiol Infect Dis 1996 Jul;15(7):588-94.
- Takai S, Fukunaga N, Ochiai S, Imai Y, Sasaki Y, Tsubaki S, Sekizaki T. Identification of intermediately virulent Rhodococcus equi isolates from pigs.. J Clin Microbiol 1996 Apr;34(4):1034-7.
- Ross TL, Balson GA, Miners JS, Smith GD, Shewen PE, Prescott JF, Yager JA. Role of CD4+, CD8+ and double negative T-cells in the protection of SCID/beige mice against respiratory challenge with Rhodococcus equi.. Can J Vet Res 1996 Jul;60(3):186-92.
- Kanaly ST, Hines SA, Palmer GH. Transfer of a CD4+ Th1 cell line to nude mice effects clearance of Rhodococcus equi from the lung.. Infect Immun 1996 Apr;64(4):1126-32.
- de la Peña-Moctezuma A, Prescott JF. A physical map of the 85 kb virulence plasmid of Rhodococcus equi 103.. Can J Vet Res 1995 Jul;59(3):229-31.
- Takai S, Watanabe Y, Ikeda T, Ozawa T, Matsukura S, Tamada Y, Tsubaki S, Sekizaki T. Virulence-associated plasmids in Rhodococcus equi.. J Clin Microbiol 1993 Jul;31(7):1726-9.
- Takai S, Iie M, Kobayashi C, Morishita T, Nishio T, Ishida T, Fujimura T, Sasaki Y, Tsubaki S. Monoclonal antibody specific to virulence-associated 15- to 17-kilodalton antigens of Rhodococcus equi.. J Clin Microbiol 1993 Oct;31(10):2780-2.
- Takai S, Sasaki Y, Ikeda T, Uchida Y, Tsubaki S, Sekizaki T. Virulence of Rhodococcus equi isolates from patients with and without AIDS.. J Clin Microbiol 1994 Feb;32(2):457-60.
- Nordmann P, Keller M, Espinasse F, Ronco E. Correlation between antibiotic resistance, phage-like particle presence, and virulence in Rhodococcus equi human isolates.. J Clin Microbiol 1994 Feb;32(2):377-83.
- Kanaly ST, Hines SA, Palmer GH. Failure of pulmonary clearance of Rhodococcus equi infection in CD4+ T-lymphocyte-deficient transgenic mice.. Infect Immun 1993 Nov;61(11):4929-32.
- Hondalus MK, Mosser DM. Survival and replication of Rhodococcus equi in macrophages.. Infect Immun 1994 Oct;62(10):4167-75.
- McNeil MM, Brown JM. The medically important aerobic actinomycetes: epidemiology and microbiology.. Clin Microbiol Rev 1994 Jul;7(3):357-417.
- Tan C, Prescott JF, Patterson MC, Nicholson VM. Molecular characterization of a lipid-modified virulence-associated protein of Rhodococcus equi and its potential in protective immunity.. Can J Vet Res 1995 Jan;59(1):51-9.
- Takai S, Ikeda T, Sasaki Y, Watanabe Y, Ozawa T, Tsubaki S, Sekizaki T. Identification of virulent Rhodococcus equi by amplification of gene coding for 15- to 17-kilodalton antigens.. J Clin Microbiol 1995 Jun;33(6):1624-7.
- Kanaly ST, Hines SA, Palmer GH. Cytokine modulation alters pulmonary clearance of Rhodococcus equi and development of granulomatous pneumonia.. Infect Immun 1995 Aug;63(8):3037-41.
- Takai S, Sekizaki T, Ozawa T, Sugawara T, Watanabe Y, Tsubaki S. Association between a large plasmid and 15- to 17-kilodalton antigens in virulent Rhodococcus equi.. Infect Immun 1991 Nov;59(11):4056-60.
- Takai S, Ohbushi S, Koike K, Tsubaki S, Oishi H, Kamada M. Prevalence of virulent Rhodococcus equi in isolates from soil and feces of horses from horse-breeding farms with and without endemic infections.. J Clin Microbiol 1991 Dec;29(12):2887-9.
- Tkachuk-Saad O, Prescott J. Rhodococcus equi plasmids: isolation and partial characterization.. J Clin Microbiol 1991 Dec;29(12):2696-700.
- Takai S, Iie M, Watanabe Y, Tsubaki S, Sekizaki T. Virulence-associated 15- to 17-kilodalton antigens in Rhodococcus equi: temperature-dependent expression and location of the antigens.. Infect Immun 1992 Jul;60(7):2995-7.
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