Assessing the Expression of Enterotoxigenic Escherichia coli-specific Surface Antigens in Recombinant Strains by Transmission Electron Microscopy and Immunolabeling
Infections with enterotoxigenic Escherichia coli (ETEC) are a major cause of travelers' diarrhea worldwide. Colonization of the small intestine mucosa is dependent on specific colonization factor antigens (CFA) and coli surface (CS) antigens. CFA/1, CS3, and CS6 are the most prevalent fimbrial...
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| Veröffentlicht in: | The journal of histochemistry and cytochemistry 2006-04, Vol.54 (4), p.473-477 |
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| creator | Ludi, Stefan Frey, Joachim Favre, Didier Stoffel, Michael H |
| description | Infections with enterotoxigenic Escherichia coli (ETEC) are a major cause of travelers' diarrhea worldwide. Colonization of the small intestine mucosa is dependent on specific colonization factor antigens (CFA) and coli surface (CS) antigens. CFA/1, CS3, and CS6 are the most prevalent fimbrial antigens found in clinical isolates. The goal of our study was to visualize the morphology of CS3 and CS6 fimbriae in wild-type and recombinant E. coli strains by means of transmission electron microscopy in conjunction with negative staining and immunolabeling. Corresponding ETEC genes were cloned into E. coli K12 strain DH10B. Expression of fimbriae was dependent on culture conditions and sample handling. Specific immunolabeling of fimbriae unequivocally demonstrated the presence of all types of surface antigens investigated. Negative staining was effective in revealing CS3 but not CS6. In addition, this technique clearly demonstrated differences in the morphology of genetically and immunologically identical CS3 surface antigens in wild-type and recombinant strains. This paper provides a basis for the assessment of recombinant vaccines. |
| doi_str_mv | 10.1369/jhc.5A6794.2005 |
| format | Article |
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Colonization of the small intestine mucosa is dependent on specific colonization factor antigens (CFA) and coli surface (CS) antigens. CFA/1, CS3, and CS6 are the most prevalent fimbrial antigens found in clinical isolates. The goal of our study was to visualize the morphology of CS3 and CS6 fimbriae in wild-type and recombinant E. coli strains by means of transmission electron microscopy in conjunction with negative staining and immunolabeling. Corresponding ETEC genes were cloned into E. coli K12 strain DH10B. Expression of fimbriae was dependent on culture conditions and sample handling. Specific immunolabeling of fimbriae unequivocally demonstrated the presence of all types of surface antigens investigated. Negative staining was effective in revealing CS3 but not CS6. In addition, this technique clearly demonstrated differences in the morphology of genetically and immunologically identical CS3 surface antigens in wild-type and recombinant strains. 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Colonization of the small intestine mucosa is dependent on specific colonization factor antigens (CFA) and coli surface (CS) antigens. CFA/1, CS3, and CS6 are the most prevalent fimbrial antigens found in clinical isolates. The goal of our study was to visualize the morphology of CS3 and CS6 fimbriae in wild-type and recombinant E. coli strains by means of transmission electron microscopy in conjunction with negative staining and immunolabeling. Corresponding ETEC genes were cloned into E. coli K12 strain DH10B. Expression of fimbriae was dependent on culture conditions and sample handling. Specific immunolabeling of fimbriae unequivocally demonstrated the presence of all types of surface antigens investigated. Negative staining was effective in revealing CS3 but not CS6. In addition, this technique clearly demonstrated differences in the morphology of genetically and immunologically identical CS3 surface antigens in wild-type and recombinant strains. This paper provides a basis for the assessment of recombinant vaccines.</description><subject>Antigens, Bacterial - biosynthesis</subject><subject>Enterotoxins - biosynthesis</subject><subject>Escherichia coli - metabolism</subject><subject>Escherichia coli K12 - genetics</subject><subject>Escherichia coli K12 - metabolism</subject><subject>Escherichia coli Proteins - biosynthesis</subject><subject>Fimbriae Proteins - biosynthesis</subject><subject>Fimbriae, Bacterial - genetics</subject><subject>Fimbriae, Bacterial - metabolism</subject><subject>Fimbriae, Bacterial - ultrastructure</subject><subject>Immunohistochemistry</subject><subject>Microscopy, Electron, Transmission</subject><subject>Recombination, Genetic</subject><issn>0022-1554</issn><issn>1551-5044</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2006</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kcGP1CAUxhujcWdXz94MFz3ZWSjQluNkU91N1pi465lQ-pgyaWGENrPz__iHStNJvBkODx4_vveRL8s-ELwltBS3h15v-a6sBNsWGPNX2YZwTnKOGXudbTAuijw12FV2HeMBY8IYr99mV6SkjNGi3mR_djFCjNbt0dQDal6OYTl6h7xBjZsg-Mm_2D04q1ETdQ_B6t4qpP1g83gEbU26eZqDURrQzk0LG5F16CdoP7bWKTehpykom9rtGT0H5eJo1yHNAHoKafPd6uCj9sczUq5DD-M4Oz-oFoZk7V32xqghwvtLvcl-fW2e7-7zxx_fHu52j7lmmE45L0UJte404XXZKRCCG2pE0RpsmKgMhaoSQvO6wLXRuDK4BqoqURYdYQZKepN9XnWPwf-eIU4y-dQwDMqBn6Msq6osRFEl8HYFF9MxgJHHYEcVzpJguQQjUzByDUYuwaQXHy_ScztC94-_JJGALysQ1R7kwc_Bpa_-R-_Tivd2359sABlHNQxJncjT6cSZTKui9C9vOakH</recordid><startdate>20060401</startdate><enddate>20060401</enddate><creator>Ludi, Stefan</creator><creator>Frey, Joachim</creator><creator>Favre, Didier</creator><creator>Stoffel, Michael H</creator><general>Histochemical Soc</general><general>SAGE Publications</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>20060401</creationdate><title>Assessing the Expression of Enterotoxigenic Escherichia coli-specific Surface Antigens in Recombinant Strains by Transmission Electron Microscopy and Immunolabeling</title><author>Ludi, Stefan ; Frey, Joachim ; Favre, Didier ; Stoffel, Michael H</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c403t-5696e8cdc1586dae995f3f92bf0f497f3e7799c58208fc07f08e3a7962d14fe63</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2006</creationdate><topic>Antigens, Bacterial - biosynthesis</topic><topic>Enterotoxins - biosynthesis</topic><topic>Escherichia coli - metabolism</topic><topic>Escherichia coli K12 - genetics</topic><topic>Escherichia coli K12 - metabolism</topic><topic>Escherichia coli Proteins - biosynthesis</topic><topic>Fimbriae Proteins - biosynthesis</topic><topic>Fimbriae, Bacterial - genetics</topic><topic>Fimbriae, Bacterial - metabolism</topic><topic>Fimbriae, Bacterial - ultrastructure</topic><topic>Immunohistochemistry</topic><topic>Microscopy, Electron, Transmission</topic><topic>Recombination, Genetic</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Ludi, Stefan</creatorcontrib><creatorcontrib>Frey, Joachim</creatorcontrib><creatorcontrib>Favre, Didier</creatorcontrib><creatorcontrib>Stoffel, Michael H</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>The journal of histochemistry and cytochemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Ludi, Stefan</au><au>Frey, Joachim</au><au>Favre, Didier</au><au>Stoffel, Michael H</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Assessing the Expression of Enterotoxigenic Escherichia coli-specific Surface Antigens in Recombinant Strains by Transmission Electron Microscopy and Immunolabeling</atitle><jtitle>The journal of histochemistry and cytochemistry</jtitle><addtitle>J Histochem Cytochem</addtitle><date>2006-04-01</date><risdate>2006</risdate><volume>54</volume><issue>4</issue><spage>473</spage><epage>477</epage><pages>473-477</pages><issn>0022-1554</issn><eissn>1551-5044</eissn><abstract>Infections with enterotoxigenic Escherichia coli (ETEC) are a major cause of travelers' diarrhea worldwide. Colonization of the small intestine mucosa is dependent on specific colonization factor antigens (CFA) and coli surface (CS) antigens. CFA/1, CS3, and CS6 are the most prevalent fimbrial antigens found in clinical isolates. The goal of our study was to visualize the morphology of CS3 and CS6 fimbriae in wild-type and recombinant E. coli strains by means of transmission electron microscopy in conjunction with negative staining and immunolabeling. Corresponding ETEC genes were cloned into E. coli K12 strain DH10B. Expression of fimbriae was dependent on culture conditions and sample handling. Specific immunolabeling of fimbriae unequivocally demonstrated the presence of all types of surface antigens investigated. Negative staining was effective in revealing CS3 but not CS6. In addition, this technique clearly demonstrated differences in the morphology of genetically and immunologically identical CS3 surface antigens in wild-type and recombinant strains. This paper provides a basis for the assessment of recombinant vaccines.</abstract><cop>Los Angeles, CA</cop><pub>Histochemical Soc</pub><pmid>16344328</pmid><doi>10.1369/jhc.5A6794.2005</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Antigens, Bacterial - biosynthesis Enterotoxins - biosynthesis Escherichia coli - metabolism Escherichia coli K12 - genetics Escherichia coli K12 - metabolism Escherichia coli Proteins - biosynthesis Fimbriae Proteins - biosynthesis Fimbriae, Bacterial - genetics Fimbriae, Bacterial - metabolism Fimbriae, Bacterial - ultrastructure Immunohistochemistry Microscopy, Electron, Transmission Recombination, Genetic |
| title | Assessing the Expression of Enterotoxigenic Escherichia coli-specific Surface Antigens in Recombinant Strains by Transmission Electron Microscopy and Immunolabeling |
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