Recognition of Mycoplasma arthritidis membrane antigens by rats and rabbits: Comparison by immunoblotting and radioimmunoprecipitation

Washburn, Leigh Rice; Ramsay, James R.; Andrews, M.

doi:10.1016/0378-1135(88)90078-8

Cited by 7 publications

(8 citation statements)

References 28 publications

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“…Antisera. Rabbit antisera were prepared against each strain as described previously (51)(52)(53)(54); the resulting antibody responses were directed largely against surface antigens. Monoclonal antibodies (MAbs) G9a and 7a (isotype immunoglobulin G2A [IgG2A]), directed against putative adhesins MAA1 and MAA2 from strain 158p10p9, respectively, were also described previously (52).…”

Section: Methodsmentioning

confidence: 99%

Comparison of Mycoplasma arthritidis strains by enzyme-linked immunosorbent assay, immunoblotting, and DNA restriction analysis

et al. 1995

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Twenty Mycoplasma arthritidis strains or isolates were compared by a combination of enzyme-linked immunosorbent assay by an antiserum adsorption technique, Western immunoblotting, and restriction analysis of chromosomal DNA. Antigenic markers that defined strains related to strains 158p10p9, PG6, and H606 were identified. In addition, restriction analysis allowed all 20 strains to be divided into six groups. Results of restriction analysis corresponded generally with antigenic similarities, although the former did not allow grouping with as fine a precision as the latter. However, intrastrain antigenic variability, which is common among many Mycoplasma species, including M. arthritidis, introduced a complicating factor into our attempts at antigenic analysis. While serologic and antigenic analyses remain useful, we recommend that they be used with caution and in combination with other techniques for identifying and characterizing new isolates and newly acquired strains. Combinations of these techniques have proven to be useful in our laboratory for quality control and for uncovering interesting relationships among strains subjected to animal passage and their less virulent antecedents and among strains originally classified as the same but obtained from different sources and maintained, sometimes for decades, in different laboratories.

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Section: Methodsmentioning

confidence: 99%

Comparison of Mycoplasma arthritidis strains by enzyme-linked immunosorbent assay, immunoblotting, and DNA restriction analysis

et al. 1995

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“…Strain-specific antisera. Female New Zealand White rabbits obtained from a local supplier were immunized with each ofthe four M. arthritidis strains as described previously (38,39). These rabbits were part of an earlier experiment on the immune response of arthritic rats and rabbits to M.…”

Section: Methodsmentioning

confidence: 99%

“…arthritidis membrane antigens (38); therefore, the immunization protocol was actually designed to induce an active infection. Briefly, each rabbit was injected intraarticularly with 1 x 109 CFU in each knee, followed by a second injection of 3 x 109 CFU by the intravenous route 5 weeks later.…”

Section: Methodsmentioning

confidence: 99%

Comparison of four Mycoplasma arthritidis strains by enzyme immunoassay, metabolism inhibition, one- and two-dimensional electrophoresis, and immunoblotting

Washburn

Hirsch

1990

J Clin Microbiol

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Four Mycoplasma arthritidis strains, two virulent (158plOp9 and 14124plO) and two avirulent (PG6 and H606), were examined for differences in their antigenic compositions. Rabbit antisera prepared against each strain were compared for their reactivities against both homologous and heterologous strains by enzyme-linked immunosorbent assay and metabolism inhibition assay. These tests confirmed a close serologic relationship among the four strains. Only by cross-absorbing each antiserum with intact cells from both homologous and heterologous strains could serologic differences be detected. Interestingly, antigenic variability was minimal among those antigens involved in the metabolism inhibition reaction, suggesting that they may be highly conserved within this species. No differences in protein composition could be detected by one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis. However, slight variation was apparent on two-dimensional gels, and antigens possessing strain-specific epitopes were detected by two-dimensional immunoblotting experiments performed with cross-absorbed antisera. The enzyme-linked immunosorbent assay, two-dimensional electrophoresis, and immunoblotting experiments showed that the two virulent strains were closely related to each other, while the avirulent strains were more distantly related to each other and to the other two strains. The relationship of M. arthritidis serologic diversity to virulence remains unclear; however, this study has demonstrated that it may be possible to subdivide M. arthritidis into serogroups on the basis of cross-absorption patterns and to identify those antigens bearing strain-specific epitopes by immunoblotting with cross-absorbed antisera.

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“…(i) pAb anti-ISR1 (Kirchhoff et al, 1983a) and (ii) pAb anti-1 5 8~1 0 were prepared against whole broth-grown cells of M. arthitidis strain ISRl and substrain 158~10, respectively, according to the protocol of Morton & Roberts (1967), as recently described (Rosengarten e t al., 1994). The preparation and characteristics of (iii) pAb anti-PG6 to M. arthritidis strain PG6 and (iv) pAb anti-H606 to strain H606 have been previously described in detail (Washburn et al, 1985(Washburn et al, ,1988Washburn & Hirsch, 1990 Colony immunoblotting. Colony immunoblots for detection of surface antigenic variants were prepared as previously described (Rosengarten & Wise, 1990Rosengarten et al, 1994).…”

Section: Labelling Of Mycoplasmas M Arthritidis Strains Substrainsmentioning

confidence: 99%

Major membrane proteins and lipoproteins as highly variable immunogenic surface components and strain-specific antigenic markers of Mycoplasma arthritidis

et al. 1995

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Ha n nover, B isc hof s hole r Damm 15,30173 Hannover, Germany Microbiology, Surface antigenic variation was investigated in Mycop/asma arthritidis, an agent that produces chronic arthritis in rats which shares several features with many mycoplasma-induced diseases and thus defines a well-characterized model system. Hyperimmune rabbit antisera (anti-ISRI, anti-PG6, anti-H606 and anti-158~10) to whole M. arthritidis organisms were used as immunological probes in Western immunoblots of four M. arthritidis prototype strains (ISRI, PG6, H606 and D263) and five rat-passaged substrains (ISRlpl, ISRlp7, ISRlp8, 1 5 8 ~ 1 0 and D263pl). Several prominent antigens were identified that varied in expression. By Triton X-114 phase fractionation and treatment of whole cells with trypsin and carboxypeptidase Y, these strain-variant antigens were shown to be integral membrane proteins with C-termini and portions of the polypeptide chains oriented outside the membrane. Western blot immunoscreening of a large number of randomly selected clonal isolates and well-established clonal lineages from stock cultures of M. arthritidis ISRlp7, 158~10, PG6 and H606 revealed an expanded repertoire of variant membrane proteins whose expression was subject to independent, reversible phase variation. Colony immunoblots of these clonal populations with a hyperimmune rabbit antiserum to a gel-purif ied variant membrane protein (P36) showed that this phase switching occurred at a high frequency lo-* per generation). Detailed immunological and biochemical characterization of the phase-variant membrane proteins demonstrated that they are: (i) antigenically related or distinct; (ii) apparently specific to particular strain populations; (iii) proteins or lipoproteins; (iv) major immunogens of M. arthritidis, recognized by serum antibodies from convalescent rat; and (v) able to undergo variation in expression during in viwo passage. Thus, M. arthritidis possesses a complex system capable of creating large repertoires of cell surface phenotypes which may affect the multiple interactions of this organism with its host and dictate its potential as a successful infectious agent and pathogen. to

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Recognition of Mycoplasma arthritidis membrane antigens by rats and rabbits: Comparison by immunoblotting and radioimmunoprecipitation

Cited by 7 publications

References 28 publications

Comparison of Mycoplasma arthritidis strains by enzyme-linked immunosorbent assay, immunoblotting, and DNA restriction analysis

Comparison of Mycoplasma arthritidis strains by enzyme-linked immunosorbent assay, immunoblotting, and DNA restriction analysis

Comparison of four Mycoplasma arthritidis strains by enzyme immunoassay, metabolism inhibition, one- and two-dimensional electrophoresis, and immunoblotting

Major membrane proteins and lipoproteins as highly variable immunogenic surface components and strain-specific antigenic markers of Mycoplasma arthritidis

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