Cytokine Gene Expression in CD4 Positive Cells of the Japanese Pufferfish, Takifugu rubripes
CD4(+) T (Th) cells are a central component of the adaptive immune response and are divided into distinct sets based on their specific cytokine production pattern. Several reports have suggested that fish possess Th subset activity similar to that of mammals. The aim of the present study was to isol...
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| description | CD4(+) T (Th) cells are a central component of the adaptive immune response and are divided into distinct sets based on their specific cytokine production pattern. Several reports have suggested that fish possess Th subset activity similar to that of mammals. The aim of the present study was to isolate CD4(+) T cells from the blood of Japanese pufferfish, Fugu rubripes, and to characterize their cytokine expression profile. We produced a specific antibody against Fugu CD4 and performed cell sorting with the magnetic activated cell sorting system. Sorted Fugu CD4(+) cells were characterized by morphology and expression analysis of cell marker genes. Fugu CD4(+) cells expressed T-cell marker genes but not macrophage or B-cell marker genes. In addition, peripheral blood lymphocytes were stimulated with lipopolysaccharide (LPS), polycytidylic acid (polyI:C), concanavalin A (ConA) prior to sorting, and then Multiplex RT-PCR was used to examine the expression of Th cytokines by the stimulated Fugu CD4(+) cells. LPS and polyI:C stimulation upregulated the expression of Th1, Th17 and Treg cytokines and downregulated the expression of Th2 cytokines. ConA stimulation upregulated the expression of all Th cytokines. These results suggest that fish exhibit the same upregulation of Th-specific cytokine expression as in mammals. |
| doi_str_mv | 10.1371/journal.pone.0066364 |
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Several reports have suggested that fish possess Th subset activity similar to that of mammals. The aim of the present study was to isolate CD4(+) T cells from the blood of Japanese pufferfish, Fugu rubripes, and to characterize their cytokine expression profile. We produced a specific antibody against Fugu CD4 and performed cell sorting with the magnetic activated cell sorting system. Sorted Fugu CD4(+) cells were characterized by morphology and expression analysis of cell marker genes. Fugu CD4(+) cells expressed T-cell marker genes but not macrophage or B-cell marker genes. In addition, peripheral blood lymphocytes were stimulated with lipopolysaccharide (LPS), polycytidylic acid (polyI:C), concanavalin A (ConA) prior to sorting, and then Multiplex RT-PCR was used to examine the expression of Th cytokines by the stimulated Fugu CD4(+) cells. LPS and polyI:C stimulation upregulated the expression of Th1, Th17 and Treg cytokines and downregulated the expression of Th2 cytokines. ConA stimulation upregulated the expression of all Th cytokines. These results suggest that fish exhibit the same upregulation of Th-specific cytokine expression as in mammals.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0066364</identifier><identifier>PMID: 23823320</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Adaptive immunity ; Analysis ; Animals ; Antigens - metabolism ; B cells ; Biology ; Blood ; Carp ; CD4 antigen ; CD4 Antigens - genetics ; CD4-Positive T-Lymphocytes - metabolism ; Concanavalin A ; Cytokines ; Cytokines - metabolism ; Cytology ; Fish ; Fishes ; Fluorescent Antibody Technique ; Gene expression ; Genes ; Helper cells ; Immune response ; Immune system ; Kinases ; Lipopolysaccharides ; Lymphocytes ; Lymphocytes B ; Lymphocytes T ; Macrophages ; Mammals ; Mitogens ; Multiplexing ; Peripheral blood ; Poly (I:C) ; Polyinosinic:polycytidylic acid ; Polymerase chain reaction ; Real-Time Polymerase Chain Reaction ; RNA, Messenger - genetics ; Stimulation ; T cells ; Takifugu rubripes ; Tetraodontiformes - immunology ; Tetraodontiformes - metabolism ; Zebrafish</subject><ispartof>PloS one, 2013-06, Vol.8 (6), p.e66364-e66364</ispartof><rights>COPYRIGHT 2013 Public Library of Science</rights><rights>2013 Kono, Korenaga. This is an open-access article distributed under the terms of the Creative Commons Attribution License: https://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2013 Kono, Korenaga 2013 Kono, Korenaga</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c758t-69b956cbd32bf69a8a6ad1ffd58743c4de3d274ca60bac976f6ff4fe84961d6b3</citedby><cites>FETCH-LOGICAL-c758t-69b956cbd32bf69a8a6ad1ffd58743c4de3d274ca60bac976f6ff4fe84961d6b3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3688880/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3688880/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,2100,2919,23857,27915,27916,53782,53784,79361,79362</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/23823320$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Moser, Muriel</contributor><creatorcontrib>Kono, Tomoya</creatorcontrib><creatorcontrib>Korenaga, Hiroki</creatorcontrib><title>Cytokine Gene Expression in CD4 Positive Cells of the Japanese Pufferfish, Takifugu rubripes</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>CD4(+) T (Th) cells are a central component of the adaptive immune response and are divided into distinct sets based on their specific cytokine production pattern. Several reports have suggested that fish possess Th subset activity similar to that of mammals. The aim of the present study was to isolate CD4(+) T cells from the blood of Japanese pufferfish, Fugu rubripes, and to characterize their cytokine expression profile. We produced a specific antibody against Fugu CD4 and performed cell sorting with the magnetic activated cell sorting system. Sorted Fugu CD4(+) cells were characterized by morphology and expression analysis of cell marker genes. Fugu CD4(+) cells expressed T-cell marker genes but not macrophage or B-cell marker genes. In addition, peripheral blood lymphocytes were stimulated with lipopolysaccharide (LPS), polycytidylic acid (polyI:C), concanavalin A (ConA) prior to sorting, and then Multiplex RT-PCR was used to examine the expression of Th cytokines by the stimulated Fugu CD4(+) cells. LPS and polyI:C stimulation upregulated the expression of Th1, Th17 and Treg cytokines and downregulated the expression of Th2 cytokines. ConA stimulation upregulated the expression of all Th cytokines. These results suggest that fish exhibit the same upregulation of Th-specific cytokine expression as in mammals.</description><subject>Adaptive immunity</subject><subject>Analysis</subject><subject>Animals</subject><subject>Antigens - metabolism</subject><subject>B cells</subject><subject>Biology</subject><subject>Blood</subject><subject>Carp</subject><subject>CD4 antigen</subject><subject>CD4 Antigens - genetics</subject><subject>CD4-Positive T-Lymphocytes - metabolism</subject><subject>Concanavalin A</subject><subject>Cytokines</subject><subject>Cytokines - metabolism</subject><subject>Cytology</subject><subject>Fish</subject><subject>Fishes</subject><subject>Fluorescent Antibody Technique</subject><subject>Gene expression</subject><subject>Genes</subject><subject>Helper cells</subject><subject>Immune response</subject><subject>Immune system</subject><subject>Kinases</subject><subject>Lipopolysaccharides</subject><subject>Lymphocytes</subject><subject>Lymphocytes B</subject><subject>Lymphocytes T</subject><subject>Macrophages</subject><subject>Mammals</subject><subject>Mitogens</subject><subject>Multiplexing</subject><subject>Peripheral blood</subject><subject>Poly (I:C)</subject><subject>Polyinosinic:polycytidylic acid</subject><subject>Polymerase chain reaction</subject><subject>Real-Time Polymerase Chain Reaction</subject><subject>RNA, Messenger - genetics</subject><subject>Stimulation</subject><subject>T cells</subject><subject>Takifugu rubripes</subject><subject>Tetraodontiformes - immunology</subject><subject>Tetraodontiformes - metabolism</subject><subject>Zebrafish</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><sourceid>DOA</sourceid><recordid>eNqNk-9r1DAYx4sobk7_A9HCQCZ4Z341ad8I4zbnyWBDp6-EkKTJXW69pibp2P57c143rrIXNtCW5PN8n-Sb58my1xBMIWbw48r1vhXNtHOtngJAKabkSbYPK4wmFAH8dOd_L3sRwgqAApeUPs_2EC4RxgjsZ79md9Fd21bnZzq9Tm87r0Owrs1tm89OSH7pgo32Rucz3TQhdyaPS51_FZ1oddD5ZW-M9saG5Yf8Slxb0y_63PfS206Hl9kzI5qgXw3fg-zH59Or2ZfJ-cXZfHZ8PlGsKOOEVrIqqJI1RtLQSpSCihoaUxclI1iRWuMaMaIEBVKoilFDjSFGl6SisKYSH2Rvt7pd4wIfjAkcYlphCAqCEjHfErUTK955uxb-jjth-d8J5xdc-GhVozkBSFIBECkoJoJAISVUTEJoWMFEVSatT0O2Xq51rXQbvWhGouOV1i75wt1wTMv0gCRwNAh497vXIfK1DSrZmyx1fdo3qyAraEFgQg__QR8_3UAtRDqAbY1LedVGlB8TViJUILRJO32ESqPWa6tSFRmb5kcB70cBiYn6Ni5EHwKff__2_-zFzzH7boddatHEZXBNH1PZhTFItqDyLgSvzYPJEPBNE9y7wTdNwIcmSGFvdi_oIei-6vEf8REA4Q</recordid><startdate>20130618</startdate><enddate>20130618</enddate><creator>Kono, Tomoya</creator><creator>Korenaga, Hiroki</creator><general>Public Library of Science</general><general>Public Library of Science (PLoS)</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>IOV</scope><scope>ISR</scope><scope>3V.</scope><scope>7QG</scope><scope>7QL</scope><scope>7QO</scope><scope>7RV</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TG</scope><scope>7TM</scope><scope>7U9</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>D1I</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB.</scope><scope>KB0</scope><scope>KL.</scope><scope>L6V</scope><scope>LK8</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>M7S</scope><scope>NAPCQ</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PATMY</scope><scope>PDBOC</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PTHSS</scope><scope>PYCSY</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope></search><sort><creationdate>20130618</creationdate><title>Cytokine Gene Expression in CD4 Positive Cells of the Japanese Pufferfish, Takifugu rubripes</title><author>Kono, Tomoya ; Korenaga, Hiroki</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c758t-69b956cbd32bf69a8a6ad1ffd58743c4de3d274ca60bac976f6ff4fe84961d6b3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>Adaptive immunity</topic><topic>Analysis</topic><topic>Animals</topic><topic>Antigens - 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genetics</topic><topic>Stimulation</topic><topic>T cells</topic><topic>Takifugu rubripes</topic><topic>Tetraodontiformes - immunology</topic><topic>Tetraodontiformes - metabolism</topic><topic>Zebrafish</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kono, Tomoya</creatorcontrib><creatorcontrib>Korenaga, Hiroki</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Gale In Context: Opposing Viewpoints</collection><collection>Gale In Context: Science</collection><collection>ProQuest Central (Corporate)</collection><collection>Animal Behavior Abstracts</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Nursing & Allied Health Database</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Immunology Abstracts</collection><collection>Meteorological & Geoastrophysical Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Agricultural Science Collection</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Public Health Database</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Technology Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>Materials Science & Engineering Collection</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central UK/Ireland</collection><collection>Advanced Technologies & Aerospace Collection</collection><collection>Agricultural & Environmental Science Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Technology Collection</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Materials Science Collection</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Materials Science Database</collection><collection>Nursing & Allied Health Database (Alumni Edition)</collection><collection>Meteorological & Geoastrophysical Abstracts - 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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>Kono, Tomoya</au><au>Korenaga, Hiroki</au><au>Moser, Muriel</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Cytokine Gene Expression in CD4 Positive Cells of the Japanese Pufferfish, Takifugu rubripes</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2013-06-18</date><risdate>2013</risdate><volume>8</volume><issue>6</issue><spage>e66364</spage><epage>e66364</epage><pages>e66364-e66364</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>CD4(+) T (Th) cells are a central component of the adaptive immune response and are divided into distinct sets based on their specific cytokine production pattern. Several reports have suggested that fish possess Th subset activity similar to that of mammals. The aim of the present study was to isolate CD4(+) T cells from the blood of Japanese pufferfish, Fugu rubripes, and to characterize their cytokine expression profile. We produced a specific antibody against Fugu CD4 and performed cell sorting with the magnetic activated cell sorting system. Sorted Fugu CD4(+) cells were characterized by morphology and expression analysis of cell marker genes. Fugu CD4(+) cells expressed T-cell marker genes but not macrophage or B-cell marker genes. In addition, peripheral blood lymphocytes were stimulated with lipopolysaccharide (LPS), polycytidylic acid (polyI:C), concanavalin A (ConA) prior to sorting, and then Multiplex RT-PCR was used to examine the expression of Th cytokines by the stimulated Fugu CD4(+) cells. LPS and polyI:C stimulation upregulated the expression of Th1, Th17 and Treg cytokines and downregulated the expression of Th2 cytokines. ConA stimulation upregulated the expression of all Th cytokines. These results suggest that fish exhibit the same upregulation of Th-specific cytokine expression as in mammals.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>23823320</pmid><doi>10.1371/journal.pone.0066364</doi><tpages>e66364</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Adaptive immunity Analysis Animals Antigens - metabolism B cells Biology Blood Carp CD4 antigen CD4 Antigens - genetics CD4-Positive T-Lymphocytes - metabolism Concanavalin A Cytokines Cytokines - metabolism Cytology Fish Fishes Fluorescent Antibody Technique Gene expression Genes Helper cells Immune response Immune system Kinases Lipopolysaccharides Lymphocytes Lymphocytes B Lymphocytes T Macrophages Mammals Mitogens Multiplexing Peripheral blood Poly (I:C) Polyinosinic:polycytidylic acid Polymerase chain reaction Real-Time Polymerase Chain Reaction RNA, Messenger - genetics Stimulation T cells Takifugu rubripes Tetraodontiformes - immunology Tetraodontiformes - metabolism Zebrafish |
| title | Cytokine Gene Expression in CD4 Positive Cells of the Japanese Pufferfish, Takifugu rubripes |
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