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Kim S Wise - One of the best experts on this subject based on the ideXlab platform.
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Gene Families Encoding Phase- and Size-Variable Surface Lipoproteins of Mycoplasma hyorhinis
Journal of bacteriology, 2000Co-Authors: Christine Citti, Robyn Watson-mckown, Martina Droesse, Kim S WiseAbstract:A prototype family of seven genes encoding the variable surface lipoproteins (Vlps) of Mycoplasma hyorhinis is characterized in the pathogenic SK76 strain, using long-range PCR to amplify and analyze the single chromosomal region containing expressed genes vlpA to -G, each of which is subject to phase and size variation. Smaller families of vlp genes in subclones of SK76 or in another strain of M. hyorhinis, GDL, can be attributed to deletions of specific vlp genes from the prototype array described here. Two genes, vlpA and the newly revealed vlpG, contain repeat motifs in their 3' coding regions that differ from the short tandem repeats in other vlp genes yet retain structural features common to all vlp gene products. SK76 and GDL vlp gene families are similarly organized and show sequence similarity between corresponding individual vlp genes. In light of the extensive potential for diversity within the vlp gene system, such conservation provides a provisional basis to hypothesize that vlp genes may exist in specific arrays that endow selected functions while retaining common structural features required during phase-variable expression of this set of gene products.
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Immunologic cross-reaction between HIV type 1 p17 and Mycoplasma hyorhinis variable lipoprotein.
AIDS Research and Human Retroviruses, 1998Co-Authors: Seth H. Pincus, Robert L. Cole, Robyn Watson-mckown, William J. Honnen, Barry C. Cole, Abraham Pinter, Kim S WiseAbstract:ABSTRACT Monoclonal antibodies directed against the HIV-1 matrix protein p17 that react with a component present on the surface of HIV-1-infected cells have previously been described. In this study we show that one of these monoclonal antibodies binds to persistently HIV-1-infected cell lines that are coinfected with Mycoplasma hyorhinis, but not to cell lines that are uninfected with Mycoplasma. Mycoplasma-infected cells secrete HIV-1 at a higher rate, have a slight increase in cell surface expression of gp120 and gp41, and are less sensitive to immunotoxins than uninfected cells. The anti-p17 antibody binds to a protein of M. hyorhinis grown in cell-free culture. The variable expression and size of the protein among strains is typical of the variable lipoprotein (Vlp) system of M. hyorhinis. Confirmation of the reactivity of the antibody with a Vlp was provided by demonstrating its specific binding to recombinant VlpF expressed in E. coli, and to a synthetic peptide representing the carboxy-terminal reg...
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Immunologic cross-reaction between HIV type 1 p17 and Mycoplasma hyorhinis variable lipoprotein.
AIDS Research and Human Retroviruses, 1998Co-Authors: Seth H. Pincus, Robert L. Cole, Robyn Watson-mckown, William J. Honnen, Barry C. Cole, Abraham Pinter, Kim S WiseAbstract:ABSTRACT Monoclonal antibodies directed against the HIV-1 matrix protein p17 that react with a component present on the surface of HIV-1-infected cells have previously been described. In this study we show that one of these monoclonal antibodies binds to persistently HIV-1-infected cell lines that are coinfected with Mycoplasma hyorhinis, but not to cell lines that are uninfected with Mycoplasma. Mycoplasma-infected cells secrete HIV-1 at a higher rate, have a slight increase in cell surface expression of gp120 and gp41, and are less sensitive to immunotoxins than uninfected cells. The anti-p17 antibody binds to a protein of M. hyorhinis grown in cell-free culture. The variable expression and size of the protein among strains is typical of the variable lipoprotein (Vlp) system of M. hyorhinis. Confirmation of the reactivity of the antibody with a Vlp was provided by demonstrating its specific binding to recombinant VlpF expressed in E. coli, and to a synthetic peptide representing the carboxy-terminal reg...
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Processing and surface presentation of the Mycoplasma hyorhinis variant lipoprotein VlpC.
Journal of bacteriology, 1994Co-Authors: C M Cleavinger, Mary F. Kim, Kim S WiseAbstract:The variant surface lipoprotein VlpC of Mycoplasma hyorhinis was shown to be processed by cleavage of a characteristic prokaryotic prolipoprotein signal peptide. In addition, a vlpC::phoA fusion protein expressed and translocated in Escherichia coli was recognized by surface-binding monoclonal antibodies, which identified the characteristic region II of Vlps, containing divergent external sequences proximal to the membrane, as an exposed portion of these surface proteins subject to immune recognition and selection.
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Antigenic variation in Mycoplasma hyorhinis: increased repertoire of variable lipoproteins expanding surface diversity and structural complexity.
Infection and immunity, 1993Co-Authors: R Rosengarten, P M Theiss, David Yogev, Kim S WiseAbstract:VlpE is characterized as a new member in a family of variable surface lipoproteins (Vlps) of Mycoplasma hyorhinis. VlpE shows phenotypic variation in expression and size within isogenic lineages of some strains but is absent from lineages of other strains that express only three previously known Vlps. Expression of four Vlps in some cells further indicates the presence and usage of an expanded reservoir of Vlp coding sequences, which greatly increases the capacity for surface diversification.
Cheng-chao Shou - One of the best experts on this subject based on the ideXlab platform.
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Detection of Mycoplasma hyorhinis Infection in Ovarian Cancer with in situ Hybridization and Immunohistochemistry DOI 10.1007/s11805-010-0534-8
2016Co-Authors: Hua Yang, Jian-zhi Zhang, Cheng-chao ShouAbstract:OBJECTIVE To detect Mycoplasma hyorhinis in ovarian cancer tissues and the relationship between Mycoplasma infection and ovarian cancer. METHODS All specimens obtained from 109 cases with ovarian cancer were fixed in freshly prepared 10 % neutral buffered formalin, embedded in paraffin, and cut into 4-μm sections for insitu hybridization (ISH) and then detected with immunohistochemistry (IHC). The expressions of 16S rRNA and P37 protein from Mycoplasma hyorhinis were detected respectively using ISH and IHC. SPSS 13.0 soĞ ware was employed to analyze the relationship between the results of the study and clinical pathological materials. RESULTS The expression rate of Mycoplasma hyorhinis 16S rRNA gene and P37 protein was 20.2 % (22/109) and 43.1 % (47/109 cases) in ovarian cancer tissues, respectively, but it was 0 (0/30 cases) in the normal ovarian tissues. The diff erence in Mycoplasma infection ratio between ovarian cancer tissues and normal tissues was extremely significant (P < 0.001). Anyhow, we didn’t found any association between the Mycoplasma infection and clinical pathological characters. CONCLUSION There was a Mycoplasma infection in ovarian cancer tissues, which may play a role in oncogenesis of ovarian cancer
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Mycoplasma hyorhinis: a potential risk factor in gastric cancer progression
Cancer Cell & Microenvironment, 2015Co-Authors: Hongying Duan, Cheng-chao ShouAbstract:Persistent infection of Mycoplasma hyorhinis (M. hyorhinis) is associated with various types of cancer. However, to date, there is few study identifying host factors mediating its infection and its effect on cancer patients’ prognosis also remains unknown. Recently, we reported for the first time that M. hyorhinis infects mammalian cells and promotes gastric cancer cell invasiveness via its membrane protein p37, and host Annexin A2 (ANXA2) is the cell receptor mediating M. hyorhinis infection. Downstream of ANXA2, NF-kB pathway is activated and mediates M. hyorhinis -driven cell migration. Furthermore, we demonstrated that M. hyorhinis p37 protein expression in gastric cancer tissues positively correlates with tumor metastasis and predicts poor survival. In conclusion, our study uncovers the mechanism by which M. hyorhinis infects mammalian cells and promotes cancer cell migration and unveils the effect of M. hyorhinis infection on gastric cancer survival.
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Mycoplasma hyorhinis infection promotes nf κb dependent migration of gastric cancer cells
Cancer Research, 2014Co-Authors: Hongying Duan, Hua Yang, Ling Chen, Sonya Wei Song, Yong Han, Wanyuan Chen, Cheng-chao ShouAbstract:Chronic infection of Mycoplasma hyorhinis ( M. hyorhinis ) has been postulated to be associated with several types of cancer, but its effect on patients9 survival and host factors mediating its infection remain unclear. Herein, we demonstrated that M. hyorhinis p37 protein expression in gastric cancer tissues predicts poor survival and associates with metastasis. M. hyorhinis infects mammalian cells and promotes gastric cancer cell invasiveness via its membrane protein p37. Synthesized peptide corresponding to the N -terminus of p37 prevents M. hyorhinis infection. Host Annexin A2 (ANXA2) interacts with the N -terminus of p37. In addition, EGFR forms a complex with p37 and ANXA2, and is required for M. hyorhinis –induced phosphorylation and membrane recruitment of ANXA2. M. hyorhinis infection is inhibited by siRNA-mediated knockdown of ANXA2 or EGFR, but is enhanced by expression of ectopic ANXA2 or EGFR. Downstream of ANXA2 and EGFR, the NF-κB pathway is activated and mediates M. hyorhinis –driven cell migration. In conclusion, our study unveils the effect of M. hyorhinis infection on gastric cancer survival and uncovers the mechanisms by which M. hyorhinis infects mammalian cells and promotes cancer cell migration. Cancer Res; 74(20); 5782–94. ©2014 AACR .
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Mycoplasma hyorhinis induces epithelial-mesenchymal transition in gastric cancer cell MGC803 via TLR4-NF-κB signaling.
Cancer letters, 2014Co-Authors: Hongying Duan, Cheng-chao ShouAbstract:Our previous works showed chronic infection of Mycoplasma hyorhinis (M. hyorhinis) was associated with gastric cancer metastasis, but the mechanisms were unknown. Herein, we found M. hyorhinis induced epithelial-mesenchymal transition (EMT) in gastric cancer cell MGC803, which was counteracted by inhibitor of NF-κB signaling or p65 knockdown. Furthermore, we found that TLR4 associated with p37, a membrane protein of M. hyorhinis. Knock-down or inhibition of TLR4 antagonized M. hyorhinis-induced NF-κB signaling, EMT, and cell migration. Thus, M. hyorhinis induces EMT and promotes cell migration via TLR4-NF-κB signaling, which provides a clue to the pathogenesis of M. hyorhinis in gastric cancer.
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Detection of Mycoplasma hyorhinis infection in ovarian cancer with in situ hybridization and immunohistochemistry
Clinical Oncology and Cancer Research, 2010Co-Authors: Hua Yang, Jian-zhi Zhang, Cheng-chao ShouAbstract:Objective To detect Mycoplasma hyorhinis in ovarian cancer tissues and the relationship between Mycoplasma infection and ovarian cancer.
Robyn Watson-mckown - One of the best experts on this subject based on the ideXlab platform.
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Gene Families Encoding Phase- and Size-Variable Surface Lipoproteins of Mycoplasma hyorhinis
Journal of bacteriology, 2000Co-Authors: Christine Citti, Robyn Watson-mckown, Martina Droesse, Kim S WiseAbstract:A prototype family of seven genes encoding the variable surface lipoproteins (Vlps) of Mycoplasma hyorhinis is characterized in the pathogenic SK76 strain, using long-range PCR to amplify and analyze the single chromosomal region containing expressed genes vlpA to -G, each of which is subject to phase and size variation. Smaller families of vlp genes in subclones of SK76 or in another strain of M. hyorhinis, GDL, can be attributed to deletions of specific vlp genes from the prototype array described here. Two genes, vlpA and the newly revealed vlpG, contain repeat motifs in their 3' coding regions that differ from the short tandem repeats in other vlp genes yet retain structural features common to all vlp gene products. SK76 and GDL vlp gene families are similarly organized and show sequence similarity between corresponding individual vlp genes. In light of the extensive potential for diversity within the vlp gene system, such conservation provides a provisional basis to hypothesize that vlp genes may exist in specific arrays that endow selected functions while retaining common structural features required during phase-variable expression of this set of gene products.
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Immunologic cross-reaction between HIV type 1 p17 and Mycoplasma hyorhinis variable lipoprotein.
AIDS Research and Human Retroviruses, 1998Co-Authors: Seth H. Pincus, Robert L. Cole, Robyn Watson-mckown, William J. Honnen, Barry C. Cole, Abraham Pinter, Kim S WiseAbstract:ABSTRACT Monoclonal antibodies directed against the HIV-1 matrix protein p17 that react with a component present on the surface of HIV-1-infected cells have previously been described. In this study we show that one of these monoclonal antibodies binds to persistently HIV-1-infected cell lines that are coinfected with Mycoplasma hyorhinis, but not to cell lines that are uninfected with Mycoplasma. Mycoplasma-infected cells secrete HIV-1 at a higher rate, have a slight increase in cell surface expression of gp120 and gp41, and are less sensitive to immunotoxins than uninfected cells. The anti-p17 antibody binds to a protein of M. hyorhinis grown in cell-free culture. The variable expression and size of the protein among strains is typical of the variable lipoprotein (Vlp) system of M. hyorhinis. Confirmation of the reactivity of the antibody with a Vlp was provided by demonstrating its specific binding to recombinant VlpF expressed in E. coli, and to a synthetic peptide representing the carboxy-terminal reg...
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Immunologic cross-reaction between HIV type 1 p17 and Mycoplasma hyorhinis variable lipoprotein.
AIDS Research and Human Retroviruses, 1998Co-Authors: Seth H. Pincus, Robert L. Cole, Robyn Watson-mckown, William J. Honnen, Barry C. Cole, Abraham Pinter, Kim S WiseAbstract:ABSTRACT Monoclonal antibodies directed against the HIV-1 matrix protein p17 that react with a component present on the surface of HIV-1-infected cells have previously been described. In this study we show that one of these monoclonal antibodies binds to persistently HIV-1-infected cell lines that are coinfected with Mycoplasma hyorhinis, but not to cell lines that are uninfected with Mycoplasma. Mycoplasma-infected cells secrete HIV-1 at a higher rate, have a slight increase in cell surface expression of gp120 and gp41, and are less sensitive to immunotoxins than uninfected cells. The anti-p17 antibody binds to a protein of M. hyorhinis grown in cell-free culture. The variable expression and size of the protein among strains is typical of the variable lipoprotein (Vlp) system of M. hyorhinis. Confirmation of the reactivity of the antibody with a Vlp was provided by demonstrating its specific binding to recombinant VlpF expressed in E. coli, and to a synthetic peptide representing the carboxy-terminal reg...
Seth H. Pincus - One of the best experts on this subject based on the ideXlab platform.
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Immunologic cross-reaction between HIV type 1 p17 and Mycoplasma hyorhinis variable lipoprotein.
AIDS Research and Human Retroviruses, 1998Co-Authors: Seth H. Pincus, Robert L. Cole, Robyn Watson-mckown, William J. Honnen, Barry C. Cole, Abraham Pinter, Kim S WiseAbstract:ABSTRACT Monoclonal antibodies directed against the HIV-1 matrix protein p17 that react with a component present on the surface of HIV-1-infected cells have previously been described. In this study we show that one of these monoclonal antibodies binds to persistently HIV-1-infected cell lines that are coinfected with Mycoplasma hyorhinis, but not to cell lines that are uninfected with Mycoplasma. Mycoplasma-infected cells secrete HIV-1 at a higher rate, have a slight increase in cell surface expression of gp120 and gp41, and are less sensitive to immunotoxins than uninfected cells. The anti-p17 antibody binds to a protein of M. hyorhinis grown in cell-free culture. The variable expression and size of the protein among strains is typical of the variable lipoprotein (Vlp) system of M. hyorhinis. Confirmation of the reactivity of the antibody with a Vlp was provided by demonstrating its specific binding to recombinant VlpF expressed in E. coli, and to a synthetic peptide representing the carboxy-terminal reg...
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Immunologic cross-reaction between HIV type 1 p17 and Mycoplasma hyorhinis variable lipoprotein.
AIDS Research and Human Retroviruses, 1998Co-Authors: Seth H. Pincus, Robert L. Cole, Robyn Watson-mckown, William J. Honnen, Barry C. Cole, Abraham Pinter, Kim S WiseAbstract:ABSTRACT Monoclonal antibodies directed against the HIV-1 matrix protein p17 that react with a component present on the surface of HIV-1-infected cells have previously been described. In this study we show that one of these monoclonal antibodies binds to persistently HIV-1-infected cell lines that are coinfected with Mycoplasma hyorhinis, but not to cell lines that are uninfected with Mycoplasma. Mycoplasma-infected cells secrete HIV-1 at a higher rate, have a slight increase in cell surface expression of gp120 and gp41, and are less sensitive to immunotoxins than uninfected cells. The anti-p17 antibody binds to a protein of M. hyorhinis grown in cell-free culture. The variable expression and size of the protein among strains is typical of the variable lipoprotein (Vlp) system of M. hyorhinis. Confirmation of the reactivity of the antibody with a Vlp was provided by demonstrating its specific binding to recombinant VlpF expressed in E. coli, and to a synthetic peptide representing the carboxy-terminal reg...
Guoqing Shao - One of the best experts on this subject based on the ideXlab platform.
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Development of oriC-plasmids for use in Mycoplasma hyorhinis.
Scientific reports, 2017Co-Authors: Hassan Z. A. Ishag, Maojun Liu, Qiyan Xiong, Zhixin Feng, Guoqing ShaoAbstract:Mycoplasma hyorhinis (M. hyorhinis) is an opportunistic pig pathogen, belonging to the class Mollicutes. It causes polyserositis, arthritis and cancers in vitro, increasing attention of the researchers. Currently, there is no available genetic tool to manipulate its genome. This study describes a development of oriC-plasmids harboring either large (pGEMT-LoriC) or minimum (pGEMT-MoriC) origin of replication (oriC) of M. hyorhinis along with tetracycline resistance marker.These plasmids were successfully transformed into M. hyorhinis with average transformation frequency of 1.5 × 10−4 and 2.0 × 10−5 transformants/CFU for pGEMT-LoriC and pGEMT-MoriC respectively, and were integrated at the chromosomal oriC as well as remained freely replicating. We also constructed a Mini-oriC-HT1 targeting plasmid by inclusion of hlyC arms and was used to inactivate hlyC at average frequency of 50%. The efficiency of hlyC inactivation was further improved (by 90%) when Mini-oriC-HT2 that contains E. coli recA was used. In both cases, hemolysin mutant bacteria diminished the ability to lyse mouse RBCs compared to wild-type (P
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development of oric plasmids for use in Mycoplasma hyorhinis
Scientific Reports, 2017Co-Authors: Hassan Z. A. Ishag, Maojun Liu, Qiyan Xiong, Zhixin Feng, Guoqing ShaoAbstract:Mycoplasma hyorhinis (M. hyorhinis) is an opportunistic pig pathogen, belonging to the class Mollicutes. It causes polyserositis, arthritis and cancers in vitro, increasing attention of the researchers. Currently, there is no available genetic tool to manipulate its genome. This study describes a development of oriC-plasmids harboring either large (pGEMT-LoriC) or minimum (pGEMT-MoriC) origin of replication (oriC) of M. hyorhinis along with tetracycline resistance marker.These plasmids were successfully transformed into M. hyorhinis with average transformation frequency of 1.5 × 10−4 and 2.0 × 10−5 transformants/CFU for pGEMT-LoriC and pGEMT-MoriC respectively, and were integrated at the chromosomal oriC as well as remained freely replicating. We also constructed a Mini-oriC-HT1 targeting plasmid by inclusion of hlyC arms and was used to inactivate hlyC at average frequency of 50%. The efficiency of hlyC inactivation was further improved (by 90%) when Mini-oriC-HT2 that contains E. coli recA was used. In both cases, hemolysin mutant bacteria diminished the ability to lyse mouse RBCs compared to wild-type (P < 0.001). OriC-plasmids described in this study may, therefore open the way for functional genomics in M. hyorhinis. Furthermore, this is a first study demonstrated the gene associated with a hemolytic phenotype in Mycoplasmas.
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e coli reca gene improves gene targeted homologous recombination in Mycoplasma hyorhinis
Journal of Microbiological Methods, 2017Co-Authors: Hassan Z. A. Ishag, Maojun Liu, Qiyan Xiong, Zhixin Feng, Guoqing ShaoAbstract:Mycoplasma hyorhinis is an opportunistic pathogen of pigs. Recently, it has been shown to transform cell cultures, increasing the attention of the researchers. Studies on the pathogenesis require specific genetic tool that is not yet available for the pathogen. To address this limitation, we constructed two suicide plasmids pGEMT-tetM/LR and pGEMT-recA-tetM/LR having a tetracycline resistance marker flanked by two hemolysin gene arms. The latter plasmid encodes an E. coli recA, a gene involved in DNA recombination, repair and maintenance of DNA. Using inactivation of the hemolysin gene, which results in a detectable and measurable phenotype, we found that each plasmid can disrupt the hemolysin gene of M. hyorhinis through a double cross-over homologous recombination. However, inclusion of the E. coli recA gene in the construct resulted in 9-fold increase in the frequency of hemolysin gene mutants among the screened tetracycline resistance colonies. The resultant hemolysin mutant strain lacks the ability to lyse mouse bed blood cells (RBC) when tested in vitro (p<0.001). The host-plasmid system described in this study, has applications for the genetic manipulation of this pathogen and potentially other Mycoplasmas.
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GFP as a marker for transient gene transfer and expression in Mycoplasma hyorhinis
SpringerPlus, 2016Co-Authors: Hassan Z. A. Ishag, Maojun Liu, Qiyan Xiong, Zhixin Feng, Ruosong Yang, Guoqing ShaoAbstract:Mycoplasma hyorhinis (M. hyorhinis) is an opportunistic pathogen of pigs and has been shown to transform cell cultures, which has increased the interest of researchers. The green florescence proteins (GFP) gene of Aquorea victoria, proved to be a vital marker to identify transformed cells in mixed populations. Use of GFP to observe gene transfer and expression in M. hyorhinis (strain HUB-1) has not been described. We have constructed a pMD18-O/MHRgfp plasmid containing the p97 gene promoter, origin of replication, tetracycline resistance marker and GFP gene controlled by the p97 gene promoter. The plasmid transformed into M. hyorhinis with a frequency of ~4 × 10−3 cfu/µg plasmid DNA and could be detected by PCR amplification of the GFP gene from the total DNA of the transformant Mycoplasmas. Analysis of a single clone grown on KM2-Agar containing tetracycline, showed a green fluorescence color. Conclusively, this report suggests the usefulness of GFP to monitor transient gene transfer and expression in M. hyorhinis, eventually minimizing screening procedures for gene transfer and expression.
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Development of a loop-mediated isothermal amplification assay for rapid detection of Mycoplasma hyorhinis.
Clinical laboratory, 2013Co-Authors: Du Gaimei, Maojun Liu, Qiyan Xiong, Fangfang Bai, Zhixin Feng, Guoqing ShaoAbstract:BACKGROUND To establish a method for sensitive and rapid diagnosis of Mycoplasma hyorhinis in clinical specimens, a simple, sensitive loop-mediated isothermal amplification (LAMP) assay was designed and evaluated. METHODS Three sets of four special primers, recognizing distinct sequences of the target, were designed for sensitive, specific amplification of nucleic acid under isothermal conditions. The LAMP assay was carried out using 35 clinical specimens of bronchoalveolar lavage fluid (BALF) from pigs. For comparison, these specimens were also tested using conventional PCR, real-time PCR, and nested PCR assays. RESULTS After optimization of the reaction condition and reaction system, the LAMP reaction successfully detected Mycoplasma hyorhinis within 40 minutes at 61 degrees C. The LAMP assay achieved a sensitivity of 10(1) copies per microL at 61 degrees C in 40 minutes, compared to real-time PCR and nested PCR, and was over 10(3) times more sensitive than conventional PCR. In the test for the specificity of the LAMP assay, only Mycoplasma hyorhinis genomic DNA was positive and no other microorganisms were positive with the primers, indicating that the LAMP assay is specific to Mycoplasma hyorhinis. Mycoplasma hyorhinis was detected in 32 samples using the LAMP and real-time PCR assays and in 27 and 11 samples using the nested PCR assay and conventional PCR assay, respectively. All the positive samples detected by real-time PCR, nested PCR and conventional PCR assays were positive in the LAMP assay. CONCLUSIONS The LAMP assay is inexpensive, easy to perform, shows a rapid reaction and does not require complex instruments like PCR. Therefore, LAMP is a simple, accurate, fast, and economical assay suitable as an alternative in veterinary practices.
