Unbiased proteomics analysis demonstrates significant variability in mucosal immune factor expression depending on the site and method of collection
Female genital tract secretions are commonly sampled by lavage of the ectocervix and vaginal vault or via a sponge inserted into the endocervix for evaluating inflammation status and immune factors critical for HIV microbicide and vaccine studies. This study uses a proteomics approach to comprehensi...
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| creator | Birse, Kenzie M Burgener, Adam Westmacott, Garrett R McCorrister, Stuart Novak, Richard M Ball, T Blake |
| description | Female genital tract secretions are commonly sampled by lavage of the ectocervix and vaginal vault or via a sponge inserted into the endocervix for evaluating inflammation status and immune factors critical for HIV microbicide and vaccine studies. This study uses a proteomics approach to comprehensively compare the efficacy of these methods, which sample from different compartments of the female genital tract, for the collection of immune factors. Matching sponge and lavage samples were collected from 10 healthy women and were analyzed by tandem mass spectrometry. Data was analyzed by a combination of differential protein expression analysis, hierarchical clustering and pathway analysis. Of the 385 proteins identified, endocervical sponge samples collected nearly twice as many unique proteins as cervicovaginal lavage (111 vs. 61) with 55% of proteins common to both (213). Each method/site identified 73 unique proteins that have roles in host immunity according to their gene ontology. Sponge samples enriched for specific inflammation pathways including acute phase response proteins (p = 3.37×10(-24)) and LXR/RXR immune activation pathways (p = 8.82×10(-22)) while the role IL-17A in psoriasis pathway (p = 5.98×10(-4)) and the complement system pathway (p = 3.91×10(-3)) were enriched in lavage samples. Many host defense factors were differentially enriched (p |
| doi_str_mv | 10.1371/journal.pone.0079505 |
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This study uses a proteomics approach to comprehensively compare the efficacy of these methods, which sample from different compartments of the female genital tract, for the collection of immune factors. Matching sponge and lavage samples were collected from 10 healthy women and were analyzed by tandem mass spectrometry. Data was analyzed by a combination of differential protein expression analysis, hierarchical clustering and pathway analysis. Of the 385 proteins identified, endocervical sponge samples collected nearly twice as many unique proteins as cervicovaginal lavage (111 vs. 61) with 55% of proteins common to both (213). Each method/site identified 73 unique proteins that have roles in host immunity according to their gene ontology. Sponge samples enriched for specific inflammation pathways including acute phase response proteins (p = 3.37×10(-24)) and LXR/RXR immune activation pathways (p = 8.82×10(-22)) while the role IL-17A in psoriasis pathway (p = 5.98×10(-4)) and the complement system pathway (p = 3.91×10(-3)) were enriched in lavage samples. Many host defense factors were differentially enriched (p<0.05) between sites including known/potential antimicrobial factors (n = 21), S100 proteins (n = 9), and immune regulatory factors such as serpins (n = 7). Immunoglobulins (n = 6) were collected at comparable levels in abundance in each site although 25% of those identified were unique to sponge samples. This study demonstrates significant differences in types and quantities of immune factors and inflammation pathways collected by each sampling technique. Therefore, clinical studies that measure mucosal immune activation or factors assessing HIV transmission should utilize both collection methods to obtain the greatest representation of immune factors secreted into the female genital tract.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0079505</identifier><identifier>PMID: 24244515</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Acquired immune deficiency syndrome ; Activation ; Adolescent ; Adult ; AIDS ; AIDS vaccines ; Analysis ; Cervix Uteri - metabolism ; Clinical trials ; Cluster Analysis ; Clustering ; Collection ; Complement activation ; Cytokines ; Data processing ; Disease transmission ; Enrichment ; Female ; Gene Expression ; Genital tract ; HIV ; Human immunodeficiency virus ; Humans ; Immune response ; Immunity ; Immunoglobulins ; Immunologic Factors - genetics ; Immunologic Factors - immunology ; Immunologic Factors - metabolism ; Immunology ; Infections ; Inflammation ; Inflammation - metabolism ; Laboratories ; Mass spectrometry ; Mass spectroscopy ; Medical research ; Methods ; Mucosal immunity ; Mucous Membrane - immunology ; Mucous Membrane - metabolism ; Neutrophils ; Organ Specificity - genetics ; Pathogenesis ; Pathways ; Proteins ; Proteome ; Proteomics ; Proteomics - methods ; Psoriasis ; Public health ; Retinoid X receptors ; Sampling methods ; Sampling techniques ; Scientific imaging ; Secretions ; Serine proteinase inhibitors ; Serpins ; Signal Transduction ; Studies ; Vagina ; Vagina - metabolism ; Young Adult</subject><ispartof>PloS one, 2013-11, Vol.8 (11), p.e79505-e79505</ispartof><rights>COPYRIGHT 2013 Public Library of Science</rights><rights>2013 Birse et al. 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This study uses a proteomics approach to comprehensively compare the efficacy of these methods, which sample from different compartments of the female genital tract, for the collection of immune factors. Matching sponge and lavage samples were collected from 10 healthy women and were analyzed by tandem mass spectrometry. Data was analyzed by a combination of differential protein expression analysis, hierarchical clustering and pathway analysis. Of the 385 proteins identified, endocervical sponge samples collected nearly twice as many unique proteins as cervicovaginal lavage (111 vs. 61) with 55% of proteins common to both (213). Each method/site identified 73 unique proteins that have roles in host immunity according to their gene ontology. Sponge samples enriched for specific inflammation pathways including acute phase response proteins (p = 3.37×10(-24)) and LXR/RXR immune activation pathways (p = 8.82×10(-22)) while the role IL-17A in psoriasis pathway (p = 5.98×10(-4)) and the complement system pathway (p = 3.91×10(-3)) were enriched in lavage samples. Many host defense factors were differentially enriched (p<0.05) between sites including known/potential antimicrobial factors (n = 21), S100 proteins (n = 9), and immune regulatory factors such as serpins (n = 7). Immunoglobulins (n = 6) were collected at comparable levels in abundance in each site although 25% of those identified were unique to sponge samples. This study demonstrates significant differences in types and quantities of immune factors and inflammation pathways collected by each sampling technique. Therefore, clinical studies that measure mucosal immune activation or factors assessing HIV transmission should utilize both collection methods to obtain the greatest representation of immune factors secreted into the female genital tract.</description><subject>Acquired immune deficiency syndrome</subject><subject>Activation</subject><subject>Adolescent</subject><subject>Adult</subject><subject>AIDS</subject><subject>AIDS vaccines</subject><subject>Analysis</subject><subject>Cervix Uteri - metabolism</subject><subject>Clinical trials</subject><subject>Cluster Analysis</subject><subject>Clustering</subject><subject>Collection</subject><subject>Complement activation</subject><subject>Cytokines</subject><subject>Data processing</subject><subject>Disease transmission</subject><subject>Enrichment</subject><subject>Female</subject><subject>Gene Expression</subject><subject>Genital tract</subject><subject>HIV</subject><subject>Human immunodeficiency virus</subject><subject>Humans</subject><subject>Immune response</subject><subject>Immunity</subject><subject>Immunoglobulins</subject><subject>Immunologic Factors - genetics</subject><subject>Immunologic Factors - immunology</subject><subject>Immunologic Factors - metabolism</subject><subject>Immunology</subject><subject>Infections</subject><subject>Inflammation</subject><subject>Inflammation - metabolism</subject><subject>Laboratories</subject><subject>Mass spectrometry</subject><subject>Mass spectroscopy</subject><subject>Medical research</subject><subject>Methods</subject><subject>Mucosal immunity</subject><subject>Mucous Membrane - immunology</subject><subject>Mucous Membrane - metabolism</subject><subject>Neutrophils</subject><subject>Organ Specificity - genetics</subject><subject>Pathogenesis</subject><subject>Pathways</subject><subject>Proteins</subject><subject>Proteome</subject><subject>Proteomics</subject><subject>Proteomics - methods</subject><subject>Psoriasis</subject><subject>Public health</subject><subject>Retinoid X receptors</subject><subject>Sampling methods</subject><subject>Sampling techniques</subject><subject>Scientific imaging</subject><subject>Secretions</subject><subject>Serine proteinase inhibitors</subject><subject>Serpins</subject><subject>Signal Transduction</subject><subject>Studies</subject><subject>Vagina</subject><subject>Vagina - 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Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Birse, Kenzie M</au><au>Burgener, Adam</au><au>Westmacott, Garrett R</au><au>McCorrister, Stuart</au><au>Novak, Richard M</au><au>Ball, T Blake</au><au>Gray, Clive M.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Unbiased proteomics analysis demonstrates significant variability in mucosal immune factor expression depending on the site and method of collection</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2013-11-14</date><risdate>2013</risdate><volume>8</volume><issue>11</issue><spage>e79505</spage><epage>e79505</epage><pages>e79505-e79505</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>Female genital tract secretions are commonly sampled by lavage of the ectocervix and vaginal vault or via a sponge inserted into the endocervix for evaluating inflammation status and immune factors critical for HIV microbicide and vaccine studies. This study uses a proteomics approach to comprehensively compare the efficacy of these methods, which sample from different compartments of the female genital tract, for the collection of immune factors. Matching sponge and lavage samples were collected from 10 healthy women and were analyzed by tandem mass spectrometry. Data was analyzed by a combination of differential protein expression analysis, hierarchical clustering and pathway analysis. Of the 385 proteins identified, endocervical sponge samples collected nearly twice as many unique proteins as cervicovaginal lavage (111 vs. 61) with 55% of proteins common to both (213). Each method/site identified 73 unique proteins that have roles in host immunity according to their gene ontology. Sponge samples enriched for specific inflammation pathways including acute phase response proteins (p = 3.37×10(-24)) and LXR/RXR immune activation pathways (p = 8.82×10(-22)) while the role IL-17A in psoriasis pathway (p = 5.98×10(-4)) and the complement system pathway (p = 3.91×10(-3)) were enriched in lavage samples. Many host defense factors were differentially enriched (p<0.05) between sites including known/potential antimicrobial factors (n = 21), S100 proteins (n = 9), and immune regulatory factors such as serpins (n = 7). Immunoglobulins (n = 6) were collected at comparable levels in abundance in each site although 25% of those identified were unique to sponge samples. This study demonstrates significant differences in types and quantities of immune factors and inflammation pathways collected by each sampling technique. Therefore, clinical studies that measure mucosal immune activation or factors assessing HIV transmission should utilize both collection methods to obtain the greatest representation of immune factors secreted into the female genital tract.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>24244515</pmid><doi>10.1371/journal.pone.0079505</doi><tpages>e79505</tpages><oa>free_for_read</oa></addata></record> |
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| recordid | cdi_plos_journals_1458576899 |
| source | MEDLINE; DOAJ Directory of Open Access Journals; Public Library of Science (PLoS) Journals Open Access; EZB-FREE-00999 freely available EZB journals; PubMed Central; Free Full-Text Journals in Chemistry |
| subjects | Acquired immune deficiency syndrome Activation Adolescent Adult AIDS AIDS vaccines Analysis Cervix Uteri - metabolism Clinical trials Cluster Analysis Clustering Collection Complement activation Cytokines Data processing Disease transmission Enrichment Female Gene Expression Genital tract HIV Human immunodeficiency virus Humans Immune response Immunity Immunoglobulins Immunologic Factors - genetics Immunologic Factors - immunology Immunologic Factors - metabolism Immunology Infections Inflammation Inflammation - metabolism Laboratories Mass spectrometry Mass spectroscopy Medical research Methods Mucosal immunity Mucous Membrane - immunology Mucous Membrane - metabolism Neutrophils Organ Specificity - genetics Pathogenesis Pathways Proteins Proteome Proteomics Proteomics - methods Psoriasis Public health Retinoid X receptors Sampling methods Sampling techniques Scientific imaging Secretions Serine proteinase inhibitors Serpins Signal Transduction Studies Vagina Vagina - metabolism Young Adult |
| title | Unbiased proteomics analysis demonstrates significant variability in mucosal immune factor expression depending on the site and method of collection |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-25T02%3A21%3A56IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Unbiased%20proteomics%20analysis%20demonstrates%20significant%20variability%20in%20mucosal%20immune%20factor%20expression%20depending%20on%20the%20site%20and%20method%20of%20collection&rft.jtitle=PloS%20one&rft.au=Birse,%20Kenzie%20M&rft.date=2013-11-14&rft.volume=8&rft.issue=11&rft.spage=e79505&rft.epage=e79505&rft.pages=e79505-e79505&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0079505&rft_dat=%3Cgale_plos_%3EA478276984%3C/gale_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1458576899&rft_id=info:pmid/24244515&rft_galeid=A478276984&rft_doaj_id=oai_doaj_org_article_8e4bdc6eab114743a33a027428d8f5f8&rfr_iscdi=true |