Identification of major immunogenic proteins of Mycoplasma synoviae isolates

Berčič, Rebeka Lucijana; Slavec, Brigita; Lavrič, Miha; Narat, Mojca; Bidovec, A.; Dovč, Peter; Benčina, Dušan

doi:10.1016/j.vetmic.2007.07.020

Cited by 55 publications

(39 citation statements)

References 19 publications

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“…It is possible that their ~ 70-kda proteins sharing antigenic determinants were CysP (Bradley et al, 1988). however, they synthesize a number of other proteins of similar molecular mass and some of them can be crossreactive, too (Berčič et al, 2008a;Benčina et al, unpublished data).…”

Section: Discussionmentioning

confidence: 99%

“…Their cells were grown in a modified frey's broth containing 10 % porcine serum and harvested as described (Berčič et al, 2008a;dušanić et al, 2009). recombinant CysP (rCysP) was produced as described in detail in our previous study (Cizelj et al, 2011).…”

Section: Mycoplasma Antigens Recombinantmentioning

confidence: 99%

“…gallisepticum (S6 strain) haemagglutinin (previously termed pMGA 1.1) (Markham et al, 1992;1993) was immuno-affinity purified using mAb 6A10 immobilized to CnBr-sepharose (Sigma) as a specific ligand (Benčina et al, 1994;Miloševič-Berlič et al, 2000). The purified pMGA (Mw ~ 67 kda; pI ~ 5.5) reacted in immunoblots with mAbs 71 to pMGA (Markham et al, 1992), but not with mAb 3 to dnaK (Berčič et al, 2008a) which have in S6 strain similar Mr and pI (demina et al, 2009). …”

Section: Mycoplasma Antigens Recombinantmentioning

confidence: 99%

“…Appropriate antigens were applied as separate dots (2 µl) on strips of the Immobilon P membrane (Millipore) (Benčina et al, 2005). Optimal concentrations of antigens were previously determined by dIBA using reference antibodies to M. synoviae, M. gallisepticum, rCysP, rMSPB, pMGA 1.1 and to synthetic peptides representing partial CysP and nanh (Benčina et al, 1994;2001;2005;Berčič et al, 2008a;2011;Cizelj et al, 2011). So, pelleted mycoplasma cells were diluted in PBS (ph 7.4) 1:2000; proteins (rMSPB, rCysP, pMGA) at concentrations of ~ 5-10 µg/ml and synthetic peptides at a concentration of ~ 10 µg/ml.…”

Section: Enzyme Immunoassays Used To Detectmentioning

confidence: 99%

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Poultry infected with Mycoplasma gallisepticum or Mycoplasma synoviae produce antibo dies to their cysteine proteas e CysP

Cizelj¹,

Berčič²,

Dušanić³

et al. 2013

Acta Agriculturae Slovenica

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Poultry infected with Mycoplasma gallisepticum and Mycoplasma synoviae produce antibodies to their cysteine protease CysPMajor poultry pathogens, Mycoplasma gallisepticum and Mycoplasma synoviae possess cysteine protease CysP, which in vitro cleaves chicken immunoglobulin G (IgG) into fab (antigen-binding fragment) and fc (crystallisable region fragment). we used recombinant CysP of M. synoviae (rCysP) in enzyme immunoassays to detect antibodies to CysP in sera, synovial fluids, and in washings of respiratory tract and oviducts of chickens and turkeys experimentally or naturally infected with M. gallisepticum or M. synoviae. In poultry infected with M. synoviae, 70.4 % of samples contained antibodies reacting with rCysP. In birds infected with M. gallisepticum we detected CysP antibodies in 63.1 % of samples. Our data demonstrate that CysP is immunogenic for chickens and turkeys and indicate that M. gallisepticum and M. synoviae infecting chickens and turkeys synthesise CysP in vivo. This is the first study demonstrating that proteases of any Mycoplasma species can induce production of specific antibodies in the natural host.Key words: poultry / infections / immunology / immunogenicity / cysteine protease / Mycoplasma synoviae / Mycoplasma gallisepticum Perutnina ob okužbi z bakterijo Mycoplasma gallisepticum ali Mycoplasma synoviae proizvede specifična protitelesa proti njuni cisteinski proteazi CysPMycoplasma gallisepticum in Mycoplasma synoviae sta pomembna patogena mikroorganizma pri perutnini. Sintetizirata cisteinsko proteazo CysP, ki in vitro razgrajuje kokošje imunoglobuline G na fab in fc fragment. Za dokazovanje protiteles proti CysP z encimsko imunskimi testi v serumih, sinovialnih tekočinah in v izpirkih dihalnih traktov in jajcevodov ptic naravno ali eksperimentalno okuženih z M. synoviae ali M. gallisepticum, smo uporabili rekombinanten protein CysP (rCysP). Pri perutnini okuženi z M. synoviae je protitelesa proti CysP vsebovalo 70,4 % vzorcev, pri perutnini okuženi z M. gallisepticum pa smo protitelesa proti CysP dokazali pri 63,1 % vzorcih. Pridobljeni podatki potrjujejo, da je CysP za perutnino imunogen protein in nakazujejo, da M. gallisepticum in M. synoviae ob okužbi perutnine sintetizirata CysP in vivo. Z našo študijo prvič dokazujemo, da lahko mikoplazemska proteaza inducira nastanek specifičnih protiteles v naravnem gostitelju.Ključne besede: perutnina / okužbe / imunologija / imunogenost / cisteinska proteaza / mikoplazme / Mycoplasma synoviae / Mycoplasma gallisepticum

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Section: Discussionmentioning

confidence: 99%

Section: Mycoplasma Antigens Recombinantmentioning

confidence: 99%

Section: Mycoplasma Antigens Recombinantmentioning

confidence: 99%

Section: Enzyme Immunoassays Used To Detectmentioning

confidence: 99%

See 2 more Smart Citations

Poultry infected with Mycoplasma gallisepticum or Mycoplasma synoviae produce antibo dies to their cysteine proteas e CysP

Cizelj¹,

Berčič²,

Dušanić³

et al. 2013

Acta Agriculturae Slovenica

Self Cite

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“…The VlhAs and sialidase of M. synoviae are both dominant surface antigens (6). Although numerous residues inferred to experience strong diversifying selection ( Ͼ 3) occur on the exterior surface of the enzyme, strain variation in sialidase activity is not attributable to direct immune-mediated selection (24,25).…”

Section: Wvu1853mentioning

confidence: 99%

Diversity of Expressed vlhA Adhesin Sequences and Intermediate Hemagglutination Phenotypes in Mycoplasma synoviae

May

Brown

2011

J Bacteriol

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A reservoir of pseudogene alleles encoding the primary adhesin VlhA occurs in the avian pathogen Mycoplasma synoviae. Recombination between this reservoir and its single expression site was predicted to result in lineages of M. synoviae that each express a different vlhA allele as a consequence of host immune responses to those antigens. Such interstrain diversity at the vlhA expression site, including major differences in the predicted secondary structures of their expressed adhesins, was confirmed in 14 specimens of M. synoviae. Corresponding functional differences in the extent to which they agglutinated erythrocytes, a quantitative proxy for VlhA-mediated cytadherence, were also evident. There was a >20-fold difference between the highest-and lowest-agglutinating strains and a rheostatic distribution of intermediate phenotypes among the others (Tukey-Kramer honestly significant difference [HSD], P < 0.001). Coincubation with the sialic acid analog 2-deoxy-2,3-didehydro-N-acetylneuraminate inhibited hemagglutination in a pattern correlated with endogenous sialidase activity (r ‫؍‬ 0.91, P < 0.001), although not consistently to the same extent that erythrocyte pretreatment with sialidase purified from Clostridium perfringens did (P < 0.05). The striking correlation between the ranked hemagglutination and endogenous sialidase activities of these strains (Spearman's r ‫؍‬ 0.874, P < 0.001) is evidence that host-induced vlhA allele switching indirectly drives sequence diversity in the passenger sialidase gene of M. synoviae.Mycoplasma synoviae is a pathogen associated with osteoarthritis, synovitis, and respiratory tract lesions of poultry (15,19,20). Cytadherence mediated by its primary adhesin VlhA is a precursor to virulence (30). Posttranslational cleavage of full-length VlhA produces the peptides MSPA (carboxy-terminal portion of VlhA) and MSPB (amino-terminal portion) (Fig. 1A). Receptor binding and cytadherence are attributed to MSPA, while the function of MSPB remains undefined (30). Interrupted expression of the vlhA gene or pretreatment with antisera against MSPA resulted in the inability of M. synoviae colonies to adsorb erythrocytes, a proxy for VlhA-mediated in vivo cytadherence. Strains able to agglutinate erythrocytes in vitro caused synovitis at a significantly higher frequency than did variants incapable of hemagglutination (29).Transcription of vlhA occurs at a single promoter next to a large assemblage of promoterless vlhA pseudogenes constituting a 69-kb locus in the M. synoviae genome. VlhA variants result from unidirectional site-specific recombination of pseudogenes into one of five specific residues at the expression site. Sequential integration of different pseudogene sequences can further result in the formation of chimeric alleles. The selective pressure of anti-VlhA antibodies in vivo is proposed to perpetuate lineages of M. synoviae that each express a different allele of vlhA as an antigen-diversifying mechanism to evade the adaptive immune system of the host (1,30,31,32).Although th...

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Application of Proteomics to Elucidate Bacterium–Host Interactions

Smith

2011

Methods in Animal Proteomics

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Identification of major immunogenic proteins of Mycoplasma synoviae isolates

Cited by 55 publications

References 19 publications

Poultry infected with Mycoplasma gallisepticum or Mycoplasma synoviae produce antibo dies to their cysteine proteas e CysP

Poultry infected with Mycoplasma gallisepticum or Mycoplasma synoviae produce antibo dies to their cysteine proteas e CysP

Diversity of Expressed vlhA Adhesin Sequences and Intermediate Hemagglutination Phenotypes in Mycoplasma synoviae

Application of Proteomics to Elucidate Bacterium–Host Interactions

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