Mycoplasma synoviae invades non-phagocytic chicken cells in vitro

Dušanić, Daliborka; Berčič, Rebeka Lucijana; Cizelj, Ivanka; Salmič, Simona; Narat, Mojca; Benčina, Dušan

doi:10.1016/j.vetmic.2009.02.014

Cited by 52 publications

(48 citation statements)

References 29 publications

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“…Chicken chondrocytes (CCH) were isolated from hyaline cartilage as described previously (Dušanić et al, 2009) and cultured in Dulbecco's modified Eagle's medium (DMEM, Sigma-Aldrich) supplemented with 7.5% fetal bovine serum (FBS, Sigma-Aldrich) and 2.5% chicken serum (Sigma-Aldrich). Cells were cultured at 37 °C in an incubator with 5% CO 2 atmosphere.…”

Section: Cultivation Of Chicken Chondrocytesmentioning

confidence: 99%

“…Acta Veterinaria Hungarica 64, 2016 Cultivation of Mycoplasma synoviae Cultures of M. synoviae type strain WVU 1853 (Morrow et al, 1990) were cultured in modified Frey's medium at 38 °C temperature as described previously (Dušanić et al, 2009). Approximately 20 in vitro passages were performed prior to culture, for which M. synoviae in logarithmic growth phase was used for CCH infection.…”

Section: Cultivation Of Chicken Chondrocytesmentioning

confidence: 99%

“…Based upon the results of our previous studies (Dušanić et al, 2009(Dušanić et al, , 2012, M. synoviae broth culture (MS0) in logarithmic growth phase was used to infect CCH at a ratio of ~ 50 M. synoviae CFU per CCH. After 24, 48 and 72 h of incubation at 37 °C in 5% CO 2 , culture supernatants were collected and CCH were sampled by trypsinisation.…”

Section: Experimental Designmentioning

confidence: 99%

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Mycoplasma and host interaction: In vitro gene expression modulation in Mycoplasma synoviae and infected chicken chondrocytes

Cizelj

Dušanić

Benčina

et al. 2016

Acta Veterinaria Hungarica

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The complex interplay between Mycoplasma synoviae and chicken chondrocytes (CCH), which come into direct contact during infectious synovitis, has been examined at the level of gene expression. Our previous studies demonstrated a significant influence of M. synoviae on the level of CCH gene expression. Here, we show for the first time that in vitro co-cultivation of M. synoviae and CCH also induces upregulation of gene expression in this mycoplasma. We observed significantly increased expression of genes important for M. synoviae pathogenicity, including cysteine protease cysP, neuraminidase nanH, haemagglutinin vlhA, and the putative nuclease MS53_0284. Moreover, the pattern of gene expression was dependent on the infection environment. In CCH, significant changes in the expression of genes encoding catabolic enzymes of the cartilage extracellular matrix (cathepsins B, K and L, aggrecanase ADAM10, and matrix metalloproteinase MMP2) were demonstrated. Infection of CCH with M. synoviae also elevated the expression of the gene encoding peptidyl arginine deiminase, type III (PADI3), which is responsible for the post-translational citrullination of proteins.

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Section: Cultivation Of Chicken Chondrocytesmentioning

confidence: 99%

Section: Cultivation Of Chicken Chondrocytesmentioning

confidence: 99%

Section: Experimental Designmentioning

confidence: 99%

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Mycoplasma and host interaction: In vitro gene expression modulation in Mycoplasma synoviae and infected chicken chondrocytes

Cizelj

Dušanić

Benčina

et al. 2016

Acta Veterinaria Hungarica

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“…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%

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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“…Primary cultures of chicken chondrocytes (CCH) were established from three chickens as described previously (Dušanić et al, 2009). CCH from different animals were cultivated separately in Dulbecco Modified Eagles Medium (DMEM), supplemented with 7,5% fetal bovine serum (FBS) and 2,5% chicken serum (all from SigmaAldrich).…”

Section: Chicken Cellsmentioning

confidence: 99%

Setting up a gene expression study for tissue cells by method of quantitative real-time PCR

Dušanić¹,

Bolha²,

Narat³

et al. 2012

Acta Agriculturae Slovenica

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Mycoplasma synoviae invades non-phagocytic chicken cells in vitro

Cited by 52 publications

References 29 publications

Mycoplasma and host interaction: In vitro gene expression modulation in Mycoplasma synoviae and infected chicken chondrocytes

Mycoplasma and host interaction: In vitro gene expression modulation in Mycoplasma synoviae and infected chicken chondrocytes

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

Setting up a gene expression study for tissue cells by method of quantitative real-time PCR

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