A chimaeric plant virus vaccine protects mice against a bacterial infection
1 Axis Genetics plc, Babraham, Cambridge CB2 4AZ, UK 2 Department of Microbiology and Immunology, Louisiana State University Medical Center, School of Medicine in Shreveport, Shreveport, LA 71130-3932, USA Author for correspondence: William D. O. Hamilton. Tel: +44 1223 837611. Fax: +44 1223 837604....
Gespeichert in:
| Veröffentlicht in: | Microbiology (Society for General Microbiology) 1999-08, Vol.145 (8), p.2061-2067 |
|---|---|
| Hauptverfasser: | , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 2067 |
|---|---|
| container_issue | 8 |
| container_start_page | 2061 |
| container_title | Microbiology (Society for General Microbiology) |
| container_volume | 145 |
| creator | Brennan, Frank R Gilleland, Linda B Staczek, John Bendig, Mary M Hamilton, William D. O Gilleland, Harry E., Jr |
| description | 1 Axis Genetics plc, Babraham, Cambridge CB2 4AZ, UK
2 Department of Microbiology and Immunology, Louisiana State University Medical Center, School of Medicine in Shreveport, Shreveport, LA 71130-3932, USA
Author for correspondence: William D. O. Hamilton. Tel: +44 1223 837611. Fax: +44 1223 837604. e-mail: hamiltonw@axisgenetics.co.uk
ABSTRACT
The plant virus cowpea mosaic virus (CPMV) is an efficient carrier of foreign peptides for the generation of strong humoral immune responses. Peptides derived from both viruses and bacteria are strongly immunogenic when displayed on the surface of CPMV and elicit high titres of peptide-specific antibody. However, the protective effects of antibodies generated using bacterial epitopes in this system have yet to be demonstrated. In this study the ability of chimaeric virus particles (CVPs) to afford protection against bacterial infection was assessed. Immunization of outbred mice with CPMV expressing a peptide derived from outer-membrane protein F of Pseudomonas aeruginosa (CPMV-PAE5) generated high titres of P. aeruginosa -specific IgG that opsonized the bacteria for phagocytosis by human neutrophils and afforded protection upon challenge with two different immunotypes of P. aeruginosa in a model of chronic pulmonary infection. When examined 8 d after challenge, CVP-immunized mice had fewer severe lung lesions and fewer bacteria in their lungs compared to mice immunized with wild-type virus. Different levels of protection were seen with CPMV-PAE5 when Freund's or alum adjuvants were used. These studies highlight the ability of CVPs to generate protective immunity against infectious disease agents.
Keywords: cowpea mosaic virus, chimaeric virus particle, Pseudomonas aeruginosa , outer-membrane protein F, vaccine
Abbreviations: CF, cystic fibrosis; CPMV, cowpea mosaic virus; CVP, chimaeric virus particle; FCA, Freund's complete adjuvant; FD, Fisher-Devlin; FIA, Freund's incomplete adjuvant; KLH, keyhole limpet haemocyanin; NMS, normal mouse serum; OM, outer membrane; PMN, polymorphonuclear leucocyte; wt, wild-type. |
| doi_str_mv | 10.1099/13500872-145-8-2061 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_70000773</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>17380344</sourcerecordid><originalsourceid>FETCH-LOGICAL-c440t-1b5124b1dbbabf06ce2f4b7aeb1eeb17563f7d77b714a413198e40ac13f8bad73</originalsourceid><addsrcrecordid>eNqFkE1LAzEQhoMo1q9fIEgOInhYzWyym-2xiF8oeNFzmKTZNrLdrcm24r93ylb0ZkjIQJ6ZvDyMnYK4AjEeX4MshKh0noEqsirLRQk77ABUWVBdiV2qicg2yIgdpvQuBD0K2GcjEKqUoPMD9jThbh4W6GNwfNlg2_N1iKvE1-hcaD1fxq73rk98EZznOMPQpp4jt-h6asKGh7YmIHTtMdursUn-ZHsfsbe729ebh-z55f7xZvKcOaVEn4EtIFcWptairUXpfF4rq9Fb8HR0UcpaT7W2GhQqkDCuvBLoQNaVxamWR-ximEvZPlY-9WYRkvMNpffdKhktaGkt_wVBy0pIpQiUA-hil1L0tVlGkhK_DAizkW1-ZBuSbSqzkU1dZ9vxK7vw0z89g10CzrcAJodNHbF1If1yY6C9iXk5YPMwm3-G6M3Mt6Q7djZ0FNn9-fMbPbGU4g</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>17380344</pqid></control><display><type>article</type><title>A chimaeric plant virus vaccine protects mice against a bacterial infection</title><source>MEDLINE</source><source>PubMed Central</source><creator>Brennan, Frank R ; Gilleland, Linda B ; Staczek, John ; Bendig, Mary M ; Hamilton, William D. O ; Gilleland, Harry E., Jr</creator><creatorcontrib>Brennan, Frank R ; Gilleland, Linda B ; Staczek, John ; Bendig, Mary M ; Hamilton, William D. O ; Gilleland, Harry E., Jr</creatorcontrib><description>1 Axis Genetics plc, Babraham, Cambridge CB2 4AZ, UK
2 Department of Microbiology and Immunology, Louisiana State University Medical Center, School of Medicine in Shreveport, Shreveport, LA 71130-3932, USA
Author for correspondence: William D. O. Hamilton. Tel: +44 1223 837611. Fax: +44 1223 837604. e-mail: hamiltonw@axisgenetics.co.uk
ABSTRACT
The plant virus cowpea mosaic virus (CPMV) is an efficient carrier of foreign peptides for the generation of strong humoral immune responses. Peptides derived from both viruses and bacteria are strongly immunogenic when displayed on the surface of CPMV and elicit high titres of peptide-specific antibody. However, the protective effects of antibodies generated using bacterial epitopes in this system have yet to be demonstrated. In this study the ability of chimaeric virus particles (CVPs) to afford protection against bacterial infection was assessed. Immunization of outbred mice with CPMV expressing a peptide derived from outer-membrane protein F of Pseudomonas aeruginosa (CPMV-PAE5) generated high titres of P. aeruginosa -specific IgG that opsonized the bacteria for phagocytosis by human neutrophils and afforded protection upon challenge with two different immunotypes of P. aeruginosa in a model of chronic pulmonary infection. When examined 8 d after challenge, CVP-immunized mice had fewer severe lung lesions and fewer bacteria in their lungs compared to mice immunized with wild-type virus. Different levels of protection were seen with CPMV-PAE5 when Freund's or alum adjuvants were used. These studies highlight the ability of CVPs to generate protective immunity against infectious disease agents.
Keywords: cowpea mosaic virus, chimaeric virus particle, Pseudomonas aeruginosa , outer-membrane protein F, vaccine
Abbreviations: CF, cystic fibrosis; CPMV, cowpea mosaic virus; CVP, chimaeric virus particle; FCA, Freund's complete adjuvant; FD, Fisher-Devlin; FIA, Freund's incomplete adjuvant; KLH, keyhole limpet haemocyanin; NMS, normal mouse serum; OM, outer membrane; PMN, polymorphonuclear leucocyte; wt, wild-type.</description><identifier>ISSN: 1350-0872</identifier><identifier>EISSN: 1465-2080</identifier><identifier>DOI: 10.1099/13500872-145-8-2061</identifier><identifier>PMID: 10463172</identifier><language>eng</language><publisher>Reading: Soc General Microbiol</publisher><subject>alum ; Animals ; Antibodies, Bacterial - blood ; Antibody Specificity ; Bacterial Outer Membrane Proteins - chemistry ; Bacterial Vaccines - administration & dosage ; Bacterial Vaccines - immunology ; Bacteriology ; Biological and medical sciences ; Comovirus - genetics ; Comovirus - immunology ; Cowpea mosaic virus ; Female ; Fundamental and applied biological sciences. Psychology ; Humans ; Lung - pathology ; Lung Diseases - microbiology ; Mice ; Mice, Inbred ICR ; Microbiology ; Opsonin Proteins - immunology ; Peptides - genetics ; Peptides - immunology ; Pseudomonas aeruginosa ; Pseudomonas aeruginosa - chemistry ; Pseudomonas aeruginosa - immunology ; Pseudomonas Infections - prevention & control ; Recombinant Fusion Proteins - administration & dosage ; Recombinant Fusion Proteins - immunology ; Vaccination ; Vaccines, antisera, therapeutical immunoglobulins and monoclonal antibodies ; Vaccines, Synthetic - immunology</subject><ispartof>Microbiology (Society for General Microbiology), 1999-08, Vol.145 (8), p.2061-2067</ispartof><rights>1999 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c440t-1b5124b1dbbabf06ce2f4b7aeb1eeb17563f7d77b714a413198e40ac13f8bad73</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27903,27904</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=1911913$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/10463172$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Brennan, Frank R</creatorcontrib><creatorcontrib>Gilleland, Linda B</creatorcontrib><creatorcontrib>Staczek, John</creatorcontrib><creatorcontrib>Bendig, Mary M</creatorcontrib><creatorcontrib>Hamilton, William D. O</creatorcontrib><creatorcontrib>Gilleland, Harry E., Jr</creatorcontrib><title>A chimaeric plant virus vaccine protects mice against a bacterial infection</title><title>Microbiology (Society for General Microbiology)</title><addtitle>Microbiology</addtitle><description>1 Axis Genetics plc, Babraham, Cambridge CB2 4AZ, UK
2 Department of Microbiology and Immunology, Louisiana State University Medical Center, School of Medicine in Shreveport, Shreveport, LA 71130-3932, USA
Author for correspondence: William D. O. Hamilton. Tel: +44 1223 837611. Fax: +44 1223 837604. e-mail: hamiltonw@axisgenetics.co.uk
ABSTRACT
The plant virus cowpea mosaic virus (CPMV) is an efficient carrier of foreign peptides for the generation of strong humoral immune responses. Peptides derived from both viruses and bacteria are strongly immunogenic when displayed on the surface of CPMV and elicit high titres of peptide-specific antibody. However, the protective effects of antibodies generated using bacterial epitopes in this system have yet to be demonstrated. In this study the ability of chimaeric virus particles (CVPs) to afford protection against bacterial infection was assessed. Immunization of outbred mice with CPMV expressing a peptide derived from outer-membrane protein F of Pseudomonas aeruginosa (CPMV-PAE5) generated high titres of P. aeruginosa -specific IgG that opsonized the bacteria for phagocytosis by human neutrophils and afforded protection upon challenge with two different immunotypes of P. aeruginosa in a model of chronic pulmonary infection. When examined 8 d after challenge, CVP-immunized mice had fewer severe lung lesions and fewer bacteria in their lungs compared to mice immunized with wild-type virus. Different levels of protection were seen with CPMV-PAE5 when Freund's or alum adjuvants were used. These studies highlight the ability of CVPs to generate protective immunity against infectious disease agents.
Keywords: cowpea mosaic virus, chimaeric virus particle, Pseudomonas aeruginosa , outer-membrane protein F, vaccine
Abbreviations: CF, cystic fibrosis; CPMV, cowpea mosaic virus; CVP, chimaeric virus particle; FCA, Freund's complete adjuvant; FD, Fisher-Devlin; FIA, Freund's incomplete adjuvant; KLH, keyhole limpet haemocyanin; NMS, normal mouse serum; OM, outer membrane; PMN, polymorphonuclear leucocyte; wt, wild-type.</description><subject>alum</subject><subject>Animals</subject><subject>Antibodies, Bacterial - blood</subject><subject>Antibody Specificity</subject><subject>Bacterial Outer Membrane Proteins - chemistry</subject><subject>Bacterial Vaccines - administration & dosage</subject><subject>Bacterial Vaccines - immunology</subject><subject>Bacteriology</subject><subject>Biological and medical sciences</subject><subject>Comovirus - genetics</subject><subject>Comovirus - immunology</subject><subject>Cowpea mosaic virus</subject><subject>Female</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Humans</subject><subject>Lung - pathology</subject><subject>Lung Diseases - microbiology</subject><subject>Mice</subject><subject>Mice, Inbred ICR</subject><subject>Microbiology</subject><subject>Opsonin Proteins - immunology</subject><subject>Peptides - genetics</subject><subject>Peptides - immunology</subject><subject>Pseudomonas aeruginosa</subject><subject>Pseudomonas aeruginosa - chemistry</subject><subject>Pseudomonas aeruginosa - immunology</subject><subject>Pseudomonas Infections - prevention & control</subject><subject>Recombinant Fusion Proteins - administration & dosage</subject><subject>Recombinant Fusion Proteins - immunology</subject><subject>Vaccination</subject><subject>Vaccines, antisera, therapeutical immunoglobulins and monoclonal antibodies</subject><subject>Vaccines, Synthetic - immunology</subject><issn>1350-0872</issn><issn>1465-2080</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1999</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkE1LAzEQhoMo1q9fIEgOInhYzWyym-2xiF8oeNFzmKTZNrLdrcm24r93ylb0ZkjIQJ6ZvDyMnYK4AjEeX4MshKh0noEqsirLRQk77ABUWVBdiV2qicg2yIgdpvQuBD0K2GcjEKqUoPMD9jThbh4W6GNwfNlg2_N1iKvE1-hcaD1fxq73rk98EZznOMPQpp4jt-h6asKGh7YmIHTtMdursUn-ZHsfsbe729ebh-z55f7xZvKcOaVEn4EtIFcWptairUXpfF4rq9Fb8HR0UcpaT7W2GhQqkDCuvBLoQNaVxamWR-ximEvZPlY-9WYRkvMNpffdKhktaGkt_wVBy0pIpQiUA-hil1L0tVlGkhK_DAizkW1-ZBuSbSqzkU1dZ9vxK7vw0z89g10CzrcAJodNHbF1If1yY6C9iXk5YPMwm3-G6M3Mt6Q7djZ0FNn9-fMbPbGU4g</recordid><startdate>19990801</startdate><enddate>19990801</enddate><creator>Brennan, Frank R</creator><creator>Gilleland, Linda B</creator><creator>Staczek, John</creator><creator>Bendig, Mary M</creator><creator>Hamilton, William D. O</creator><creator>Gilleland, Harry E., Jr</creator><general>Soc General Microbiol</general><general>Society for General Microbiology</general><scope>IQODW</scope><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>7QL</scope><scope>7T7</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>19990801</creationdate><title>A chimaeric plant virus vaccine protects mice against a bacterial infection</title><author>Brennan, Frank R ; Gilleland, Linda B ; Staczek, John ; Bendig, Mary M ; Hamilton, William D. O ; Gilleland, Harry E., Jr</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c440t-1b5124b1dbbabf06ce2f4b7aeb1eeb17563f7d77b714a413198e40ac13f8bad73</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1999</creationdate><topic>alum</topic><topic>Animals</topic><topic>Antibodies, Bacterial - blood</topic><topic>Antibody Specificity</topic><topic>Bacterial Outer Membrane Proteins - chemistry</topic><topic>Bacterial Vaccines - administration & dosage</topic><topic>Bacterial Vaccines - immunology</topic><topic>Bacteriology</topic><topic>Biological and medical sciences</topic><topic>Comovirus - genetics</topic><topic>Comovirus - immunology</topic><topic>Cowpea mosaic virus</topic><topic>Female</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Humans</topic><topic>Lung - pathology</topic><topic>Lung Diseases - microbiology</topic><topic>Mice</topic><topic>Mice, Inbred ICR</topic><topic>Microbiology</topic><topic>Opsonin Proteins - immunology</topic><topic>Peptides - genetics</topic><topic>Peptides - immunology</topic><topic>Pseudomonas aeruginosa</topic><topic>Pseudomonas aeruginosa - chemistry</topic><topic>Pseudomonas aeruginosa - immunology</topic><topic>Pseudomonas Infections - prevention & control</topic><topic>Recombinant Fusion Proteins - administration & dosage</topic><topic>Recombinant Fusion Proteins - immunology</topic><topic>Vaccination</topic><topic>Vaccines, antisera, therapeutical immunoglobulins and monoclonal antibodies</topic><topic>Vaccines, Synthetic - immunology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Brennan, Frank R</creatorcontrib><creatorcontrib>Gilleland, Linda B</creatorcontrib><creatorcontrib>Staczek, John</creatorcontrib><creatorcontrib>Bendig, Mary M</creatorcontrib><creatorcontrib>Hamilton, William D. O</creatorcontrib><creatorcontrib>Gilleland, Harry E., Jr</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Microbiology (Society for General Microbiology)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Brennan, Frank R</au><au>Gilleland, Linda B</au><au>Staczek, John</au><au>Bendig, Mary M</au><au>Hamilton, William D. O</au><au>Gilleland, Harry E., Jr</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A chimaeric plant virus vaccine protects mice against a bacterial infection</atitle><jtitle>Microbiology (Society for General Microbiology)</jtitle><addtitle>Microbiology</addtitle><date>1999-08-01</date><risdate>1999</risdate><volume>145</volume><issue>8</issue><spage>2061</spage><epage>2067</epage><pages>2061-2067</pages><issn>1350-0872</issn><eissn>1465-2080</eissn><abstract>1 Axis Genetics plc, Babraham, Cambridge CB2 4AZ, UK
2 Department of Microbiology and Immunology, Louisiana State University Medical Center, School of Medicine in Shreveport, Shreveport, LA 71130-3932, USA
Author for correspondence: William D. O. Hamilton. Tel: +44 1223 837611. Fax: +44 1223 837604. e-mail: hamiltonw@axisgenetics.co.uk
ABSTRACT
The plant virus cowpea mosaic virus (CPMV) is an efficient carrier of foreign peptides for the generation of strong humoral immune responses. Peptides derived from both viruses and bacteria are strongly immunogenic when displayed on the surface of CPMV and elicit high titres of peptide-specific antibody. However, the protective effects of antibodies generated using bacterial epitopes in this system have yet to be demonstrated. In this study the ability of chimaeric virus particles (CVPs) to afford protection against bacterial infection was assessed. Immunization of outbred mice with CPMV expressing a peptide derived from outer-membrane protein F of Pseudomonas aeruginosa (CPMV-PAE5) generated high titres of P. aeruginosa -specific IgG that opsonized the bacteria for phagocytosis by human neutrophils and afforded protection upon challenge with two different immunotypes of P. aeruginosa in a model of chronic pulmonary infection. When examined 8 d after challenge, CVP-immunized mice had fewer severe lung lesions and fewer bacteria in their lungs compared to mice immunized with wild-type virus. Different levels of protection were seen with CPMV-PAE5 when Freund's or alum adjuvants were used. These studies highlight the ability of CVPs to generate protective immunity against infectious disease agents.
Keywords: cowpea mosaic virus, chimaeric virus particle, Pseudomonas aeruginosa , outer-membrane protein F, vaccine
Abbreviations: CF, cystic fibrosis; CPMV, cowpea mosaic virus; CVP, chimaeric virus particle; FCA, Freund's complete adjuvant; FD, Fisher-Devlin; FIA, Freund's incomplete adjuvant; KLH, keyhole limpet haemocyanin; NMS, normal mouse serum; OM, outer membrane; PMN, polymorphonuclear leucocyte; wt, wild-type.</abstract><cop>Reading</cop><pub>Soc General Microbiol</pub><pmid>10463172</pmid><doi>10.1099/13500872-145-8-2061</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1350-0872 |
| ispartof | Microbiology (Society for General Microbiology), 1999-08, Vol.145 (8), p.2061-2067 |
| issn | 1350-0872 1465-2080 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_70000773 |
| source | MEDLINE; PubMed Central |
| subjects | alum Animals Antibodies, Bacterial - blood Antibody Specificity Bacterial Outer Membrane Proteins - chemistry Bacterial Vaccines - administration & dosage Bacterial Vaccines - immunology Bacteriology Biological and medical sciences Comovirus - genetics Comovirus - immunology Cowpea mosaic virus Female Fundamental and applied biological sciences. Psychology Humans Lung - pathology Lung Diseases - microbiology Mice Mice, Inbred ICR Microbiology Opsonin Proteins - immunology Peptides - genetics Peptides - immunology Pseudomonas aeruginosa Pseudomonas aeruginosa - chemistry Pseudomonas aeruginosa - immunology Pseudomonas Infections - prevention & control Recombinant Fusion Proteins - administration & dosage Recombinant Fusion Proteins - immunology Vaccination Vaccines, antisera, therapeutical immunoglobulins and monoclonal antibodies Vaccines, Synthetic - immunology |
| title | A chimaeric plant virus vaccine protects mice against a bacterial infection |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-23T18%3A04%3A07IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=A%20chimaeric%20plant%20virus%20vaccine%20protects%20mice%20against%20a%20bacterial%20infection&rft.jtitle=Microbiology%20(Society%20for%20General%20Microbiology)&rft.au=Brennan,%20Frank%20R&rft.date=1999-08-01&rft.volume=145&rft.issue=8&rft.spage=2061&rft.epage=2067&rft.pages=2061-2067&rft.issn=1350-0872&rft.eissn=1465-2080&rft_id=info:doi/10.1099/13500872-145-8-2061&rft_dat=%3Cproquest_cross%3E17380344%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=17380344&rft_id=info:pmid/10463172&rfr_iscdi=true |