Abstract
Genes related to Mycoplasma hyopneumoniae-induced inflammation were identified using the gene-fishing technology, an improved method for identifying differentially expressed genes (DEGs) using an annealing control primer (ACP) system in RAW264.7 cells. After treatment with M. hyopneumo-niae, 16 DEGs were expressed in RAW264.7 cells using a pre-screening system. Among these 16 DEGs, 11 DEGs (DEGs 1, 4, 5–10, 12-15) were selected and sequenced directly, revealing that DEG12 (Grap), DEG14 (Gadd45), and DEG15 (secreted phosphoprotein 1) were related to inflammatory cytokines. This is the first report that intact M. hyopneumoniae induces the expression of Grap, Gadd 45β, and secreted phosphoprotein 1 in RAW264.7 cells. Subsequently, these genes may be targets for screening novel inhibitors of the mycoplasmal inflammatory response.
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Asai, T., Okada, M., Ono, M., Irisawa, T., Mori, Y., Yokomizo, Y. and Sato, S. (1993). Increased levels of tumor necrosis factor and interleukin 1 in bronchoalveolar lavage fluids from pigs infected with Mycoplasma hyopneumoniae. Vet. Immunol. Immunopathology, 38, 253–260.
Asai, T., Okada, M., Ono, M., Mori, Y., Yokomizo, Y. and Sato, S. (1994). Detection of interleukin-6 and prostaglan-din E2 in bronchoalveolar lavage fluids of pigs experimentally infected with Mycoplasma hyopneumoniae. Vet. Immunol. Immunopathol., 44, 97–102.
DeBey, M.C and Ross, R.F (1994). Ciliostasis and loss of cilia induced by Mycoplasma hyopneumoniae in porcine tracheal organ cultures. Infect. Immun., 62, 5312–5318.
Friis, N.F (1975). Some recommendations concerning primary isolation of Mycoplasma suipneumoniae and Myco-plasma floculare: a survey. Nord. Vet. Med., 27, 337–339.
Hwang, I.T, Kim, Y.J, Kim, S.H, Kwak, C.I, Gu, Y.Y and Chun, J.Y (2003). Annealing control primer system for improving specificity of PCR amplification. BioTechniques, 35, 1180–1184.
Hwang, K.C, Cui, X.S, Park, S.P, Shin, M.R, Park, S.Y, Kim, E.Y and Kim, N.H (2004). Identification of differentially regulated genes in bovine blastocysts using an annealing control primer system. Mol. Reprod. Dev., 69, 43–51.
Hwang, M.H, Lim, J.H, Yun, H.I, Kim, J.C, Jung, B.Y, Hsu, W.H and Park, S.C (2006). The effect of polyclonal antibody on intracellular calcium increase induced by Myco-plasma hyopneumoniae in porcine tracheal cells. Vet. J., 172, 556–560.
Hwang, M.H, Chang, Z.Q, Kang, E.H, Lim, J.H, Yun, H.I, Rhee, M.H, Jeong, K.S and Park, S.C (2008). Surfactin C inhibits Mycoplasma hyopneumoniae-induced transcription of proinflammatory cytokines and nitric oxide production in murine RAW 264.7 cells. Biotechnol. Lett., 30, 229–233.
Kim, Y.J, Kwak, C.I, Gu, Y.Y, Hwang, I.T and Chun, J.Y (2004). Annealing control primer system for identification of differentially expressed genes on agarose gels. BioTechniques, 36, 424–426.
Kohan, M., Bader, R., Puxeddu, I., Levi- Schaffer, F., Breuer, R. and Berkman, N. (2007). Enhanced osteopontin expression in a murine model of allergen-induced airway remodeling. Clin. Exp. Allergy, 37, 1444–1454.
Lu, B., Ferrandino, A.F and Flavell, R.A (2004). Gadd45beta is important for perpetuating cognate and inflammatory signals in T cells. Nat. Immunol., 5, 38–44.
Lu, B., Yu, H., Chow, C., Li, B., Zheng, W., Davis, R.J and Flavell, R.A (2001). GADD45 gamma mediates the activation of the p38 and JNK MAP kinase pathways and cytokine production in effector TH1 cells. Immunity, 14, 583–590.
O’Regan, A.W, Hayden, J., Body, S., Liaw, L., Mulligan, N., Goetschkes, M. and Berman, J.S (2001). Abnormal pulmonary granuloma formation in osteopontin-deficient mice. Am. J. Respir. Crit. Care. Med., 164, 2243–2247.
Park, S.C, Yibchok-Anun, S., Cheng, H., Young, T.F, Thacker, E.L., Minion, F.C, Ross, R.F and Hsu, W.H (2002). Mycoplasma hyopneumoniae increases intracellular calcium release in porcine ciliated tracheal cells. Infect. Immun., 70, 2502–2506.
Randy, S., Ouyang, Y.B, Qu, C., Alonso, A., Sperzel, L., Mustelin, T., Kaplan, M.H and Feng, G.S (2002). Grap negatively regulates T-cell receptor-elicited lymphocyte proliferation and interleukin-2 induction. Mol. Cell. Biol., 22, 3230–3236.
Serlin, D.M, Kuang, P., Subramanian, M., O’Regan, A., Li, X., Berman, J.S and Goldstein, R.H (2006). Interleukin-1beta induces osteopontin expression in pulmonary fibro-blasts. J. Cell. Biochem., 97, 519–529.
Shirai, W., Takei, M., Yoshikawa, T., Azuma, T., Kato, M., Mitamura, K., Ueki, T., Kida, A., Horie, T., Seki, N. and Sawada, S. (2004). Detection of Grb-2-related adaptor protein gene (GRAP) and peptide molecule in salivary glands of MRL/lpr mice and patients with Sjögren’s syndrome. J. Int. Med. Res., 32, 284–291.
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Hwang, MH., Choi, MJ. & Park, SC. Mycoplasma hyopneumoniae Induces Grap, Gadd45β, and secreted phosphoprotein 1 Gene Expression as Part of the Inflammatory Response in RAW264.7 Cells. Toxicol Res. 25, 119–124 (2009). https://doi.org/10.5487/TR.2009.25.3.119
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DOI: https://doi.org/10.5487/TR.2009.25.3.119