An alternative method for T-cell receptor repertoire analysis: Clustering of human V-beta subfamilies selected in responses to staphylococcal enterotoxins B and E
We have designed a convenient procedure for the analysis of Vβ repertoire expression in polyclonal T-cell populations. In this procedure T-cell RNA is converted to cDNA, polydC-tailed with terminal deoxynucleotidyl transferase and submitted to one-side specificity PCR amplification with a constant r...
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Veröffentlicht in: | Molecular immunology 1993-07, Vol.30 (10), p.877-886 |
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description | We have designed a convenient procedure for the analysis of Vβ repertoire expression in polyclonal T-cell populations. In this procedure T-cell RNA is converted to cDNA, polydC-tailed with terminal deoxynucleotidyl transferase and submitted to one-side specificity PCR amplification with a constant region oligonucleotide primer. The amplified material is then analysed by reverse spot-test hybridization: after
32P-labelling, the amplification product is put to hybridize on a membrane where specially designed Vβ subfamily-specific probes are immobilized. The radioactivity fixed on each probe can then be easily quantified and the signal obtained is directly proportional to the initial amount of homologous RNA. We applied this technique to the study of Vβ gene selection following T-cell stimulation by staphylococcal enterotoxins B and E. We show that with these toxins two almost non-overlapping sets of T-cells are recruited and that this selection is likely to be dependent on specific amino acid residues shaping the fourth complementarity determining region of the TCR-β chain. These residues constitute two tandemly-conserved tripeptide sequences (Asp
39Pro
40Gly
41)-(Val
61)Ser
70Arg
71) and (Arg
66Phe
67Ser
68)-(Asp
88Ser
89Ala
90) in the SEB- and the SEE-responsive Vβ gene clusters respectively. |
doi_str_mv | 10.1016/0161-5890(93)90011-Y |
format | Article |
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32P-labelling, the amplification product is put to hybridize on a membrane where specially designed Vβ subfamily-specific probes are immobilized. The radioactivity fixed on each probe can then be easily quantified and the signal obtained is directly proportional to the initial amount of homologous RNA. We applied this technique to the study of Vβ gene selection following T-cell stimulation by staphylococcal enterotoxins B and E. We show that with these toxins two almost non-overlapping sets of T-cells are recruited and that this selection is likely to be dependent on specific amino acid residues shaping the fourth complementarity determining region of the TCR-β chain. These residues constitute two tandemly-conserved tripeptide sequences (Asp
39Pro
40Gly
41)-(Val
61)Ser
70Arg
71) and (Arg
66Phe
67Ser
68)-(Asp
88Ser
89Ala
90) in the SEB- and the SEE-responsive Vβ gene clusters respectively.</description><identifier>ISSN: 0161-5890</identifier><identifier>EISSN: 1872-9142</identifier><identifier>DOI: 10.1016/0161-5890(93)90011-Y</identifier><identifier>PMID: 8341282</identifier><identifier>CODEN: MOIMD5</identifier><language>eng</language><publisher>Oxford: Elsevier Ltd</publisher><subject>Amino Acid Sequence ; Antigen-Antibody Reactions ; Base Sequence ; Biological and medical sciences ; Cells, Cultured ; Enterotoxins - immunology ; Fundamental and applied biological sciences. Psychology ; Genes. Genome ; Humans ; Immunoblotting ; Immunoglobulin Variable Region - chemistry ; Immunoglobulin Variable Region - genetics ; Immunoglobulin Variable Region - immunology ; Molecular and cellular biology ; Molecular genetics ; Molecular Sequence Data ; Multigene Family ; Nucleic Acid Hybridization ; Oligodeoxyribonucleotides - chemistry ; Oligonucleotide Probes ; Polymerase Chain Reaction ; Receptors, Antigen, T-Cell, alpha-beta - chemistry ; Receptors, Antigen, T-Cell, alpha-beta - immunology ; Sequence Alignment ; Sequence Homology, Amino Acid ; Staphylococcus ; Staphylococcus aureus - immunology ; T-Lymphocytes - immunology</subject><ispartof>Molecular immunology, 1993-07, Vol.30 (10), p.877-886</ispartof><rights>1993</rights><rights>1993 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c332t-ceea6a0eee9a1f2e857f9fd4cd4ac48c8cceadf5653a14dc11f5be76f29f06ad3</citedby><cites>FETCH-LOGICAL-c332t-ceea6a0eee9a1f2e857f9fd4cd4ac48c8cceadf5653a14dc11f5be76f29f06ad3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/0161-5890(93)90011-Y$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3548,27922,27923,45993</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=4868610$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/8341282$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Champagne, Eric</creatorcontrib><creatorcontrib>Huchenq, Anne</creatorcontrib><creatorcontrib>Sevin, Jacqueline</creatorcontrib><creatorcontrib>Casteran, Nathalie</creatorcontrib><creatorcontrib>Rubin, Bent</creatorcontrib><title>An alternative method for T-cell receptor repertoire analysis: Clustering of human V-beta subfamilies selected in responses to staphylococcal enterotoxins B and E</title><title>Molecular immunology</title><addtitle>Mol Immunol</addtitle><description>We have designed a convenient procedure for the analysis of Vβ repertoire expression in polyclonal T-cell populations. In this procedure T-cell RNA is converted to cDNA, polydC-tailed with terminal deoxynucleotidyl transferase and submitted to one-side specificity PCR amplification with a constant region oligonucleotide primer. The amplified material is then analysed by reverse spot-test hybridization: after
32P-labelling, the amplification product is put to hybridize on a membrane where specially designed Vβ subfamily-specific probes are immobilized. The radioactivity fixed on each probe can then be easily quantified and the signal obtained is directly proportional to the initial amount of homologous RNA. We applied this technique to the study of Vβ gene selection following T-cell stimulation by staphylococcal enterotoxins B and E. We show that with these toxins two almost non-overlapping sets of T-cells are recruited and that this selection is likely to be dependent on specific amino acid residues shaping the fourth complementarity determining region of the TCR-β chain. These residues constitute two tandemly-conserved tripeptide sequences (Asp
39Pro
40Gly
41)-(Val
61)Ser
70Arg
71) and (Arg
66Phe
67Ser
68)-(Asp
88Ser
89Ala
90) in the SEB- and the SEE-responsive Vβ gene clusters respectively.</description><subject>Amino Acid Sequence</subject><subject>Antigen-Antibody Reactions</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Cells, Cultured</subject><subject>Enterotoxins - immunology</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Genes. Genome</subject><subject>Humans</subject><subject>Immunoblotting</subject><subject>Immunoglobulin Variable Region - chemistry</subject><subject>Immunoglobulin Variable Region - genetics</subject><subject>Immunoglobulin Variable Region - immunology</subject><subject>Molecular and cellular biology</subject><subject>Molecular genetics</subject><subject>Molecular Sequence Data</subject><subject>Multigene Family</subject><subject>Nucleic Acid Hybridization</subject><subject>Oligodeoxyribonucleotides - chemistry</subject><subject>Oligonucleotide Probes</subject><subject>Polymerase Chain Reaction</subject><subject>Receptors, Antigen, T-Cell, alpha-beta - chemistry</subject><subject>Receptors, Antigen, T-Cell, alpha-beta - immunology</subject><subject>Sequence Alignment</subject><subject>Sequence Homology, Amino Acid</subject><subject>Staphylococcus</subject><subject>Staphylococcus aureus - immunology</subject><subject>T-Lymphocytes - immunology</subject><issn>0161-5890</issn><issn>1872-9142</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1993</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkc2KFDEUhYMoY8_oGyhkIaKL0qR-UikXwtiMPzDgZhRmFW4nN3akKimT1GC_jk86abvppS5CSO53z72cQ8gzzt5wxsXbcnjVyYG9GprXA2OcV7cPyIrLvq4G3tYPyeqEPCbnKf1kjAkmujNyJpuW17JekT-XnsKYMXrI7g7phHkbDLUh0ptK4zjSiBrnXN4RZ4w5uIgUPIy75NI7uh6XVLqd_0GDpdtlAk-_VxvMQNOysTC50WGiCUfUGQ11vuikOfhUfnOgKcO83Y1BB61hpOiLWMjht_OJfihzDL16Qh5ZGBM-Pd4X5NvHq5v15-r666cv68vrSjdNncuyCAIYIg7AbY2y6-1gTatNC7qVWmqNYGwnugZ4azTntttgL2w9WCbANBfk5UF3juHXgimryaW9BeAxLEn1xUjZt91_QS76QYhGFrA9gDqGlCJaNUc3QdwpztQ-Q7UPSO0DUkOj_maobkvb86P-spnQnJqOoZX6i2MdUjHNRvDapRPWSiEFZwV7f8CwmHbnMKqkHXqNpkSoszLB_XuPe0zHvUg</recordid><startdate>199307</startdate><enddate>199307</enddate><creator>Champagne, Eric</creator><creator>Huchenq, Anne</creator><creator>Sevin, Jacqueline</creator><creator>Casteran, Nathalie</creator><creator>Rubin, Bent</creator><general>Elsevier Ltd</general><general>Elsevier</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>7T5</scope><scope>H94</scope><scope>7X8</scope></search><sort><creationdate>199307</creationdate><title>An alternative method for T-cell receptor repertoire analysis: Clustering of human V-beta subfamilies selected in responses to staphylococcal enterotoxins B and E</title><author>Champagne, Eric ; Huchenq, Anne ; Sevin, Jacqueline ; Casteran, Nathalie ; Rubin, Bent</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c332t-ceea6a0eee9a1f2e857f9fd4cd4ac48c8cceadf5653a14dc11f5be76f29f06ad3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1993</creationdate><topic>Amino Acid Sequence</topic><topic>Antigen-Antibody Reactions</topic><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>Cells, Cultured</topic><topic>Enterotoxins - immunology</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Genes. Genome</topic><topic>Humans</topic><topic>Immunoblotting</topic><topic>Immunoglobulin Variable Region - chemistry</topic><topic>Immunoglobulin Variable Region - genetics</topic><topic>Immunoglobulin Variable Region - immunology</topic><topic>Molecular and cellular biology</topic><topic>Molecular genetics</topic><topic>Molecular Sequence Data</topic><topic>Multigene Family</topic><topic>Nucleic Acid Hybridization</topic><topic>Oligodeoxyribonucleotides - chemistry</topic><topic>Oligonucleotide Probes</topic><topic>Polymerase Chain Reaction</topic><topic>Receptors, Antigen, T-Cell, alpha-beta - chemistry</topic><topic>Receptors, Antigen, T-Cell, alpha-beta - immunology</topic><topic>Sequence Alignment</topic><topic>Sequence Homology, Amino Acid</topic><topic>Staphylococcus</topic><topic>Staphylococcus aureus - immunology</topic><topic>T-Lymphocytes - immunology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Champagne, Eric</creatorcontrib><creatorcontrib>Huchenq, Anne</creatorcontrib><creatorcontrib>Sevin, Jacqueline</creatorcontrib><creatorcontrib>Casteran, Nathalie</creatorcontrib><creatorcontrib>Rubin, Bent</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>Immunology Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Molecular immunology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Champagne, Eric</au><au>Huchenq, Anne</au><au>Sevin, Jacqueline</au><au>Casteran, Nathalie</au><au>Rubin, Bent</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>An alternative method for T-cell receptor repertoire analysis: Clustering of human V-beta subfamilies selected in responses to staphylococcal enterotoxins B and E</atitle><jtitle>Molecular immunology</jtitle><addtitle>Mol Immunol</addtitle><date>1993-07</date><risdate>1993</risdate><volume>30</volume><issue>10</issue><spage>877</spage><epage>886</epage><pages>877-886</pages><issn>0161-5890</issn><eissn>1872-9142</eissn><coden>MOIMD5</coden><abstract>We have designed a convenient procedure for the analysis of Vβ repertoire expression in polyclonal T-cell populations. In this procedure T-cell RNA is converted to cDNA, polydC-tailed with terminal deoxynucleotidyl transferase and submitted to one-side specificity PCR amplification with a constant region oligonucleotide primer. The amplified material is then analysed by reverse spot-test hybridization: after
32P-labelling, the amplification product is put to hybridize on a membrane where specially designed Vβ subfamily-specific probes are immobilized. The radioactivity fixed on each probe can then be easily quantified and the signal obtained is directly proportional to the initial amount of homologous RNA. We applied this technique to the study of Vβ gene selection following T-cell stimulation by staphylococcal enterotoxins B and E. We show that with these toxins two almost non-overlapping sets of T-cells are recruited and that this selection is likely to be dependent on specific amino acid residues shaping the fourth complementarity determining region of the TCR-β chain. These residues constitute two tandemly-conserved tripeptide sequences (Asp
39Pro
40Gly
41)-(Val
61)Ser
70Arg
71) and (Arg
66Phe
67Ser
68)-(Asp
88Ser
89Ala
90) in the SEB- and the SEE-responsive Vβ gene clusters respectively.</abstract><cop>Oxford</cop><pub>Elsevier Ltd</pub><pmid>8341282</pmid><doi>10.1016/0161-5890(93)90011-Y</doi><tpages>10</tpages></addata></record> |
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ispartof | Molecular immunology, 1993-07, Vol.30 (10), p.877-886 |
issn | 0161-5890 1872-9142 |
language | eng |
recordid | cdi_proquest_miscellaneous_75898745 |
source | MEDLINE; ScienceDirect Journals (5 years ago - present) |
subjects | Amino Acid Sequence Antigen-Antibody Reactions Base Sequence Biological and medical sciences Cells, Cultured Enterotoxins - immunology Fundamental and applied biological sciences. Psychology Genes. Genome Humans Immunoblotting Immunoglobulin Variable Region - chemistry Immunoglobulin Variable Region - genetics Immunoglobulin Variable Region - immunology Molecular and cellular biology Molecular genetics Molecular Sequence Data Multigene Family Nucleic Acid Hybridization Oligodeoxyribonucleotides - chemistry Oligonucleotide Probes Polymerase Chain Reaction Receptors, Antigen, T-Cell, alpha-beta - chemistry Receptors, Antigen, T-Cell, alpha-beta - immunology Sequence Alignment Sequence Homology, Amino Acid Staphylococcus Staphylococcus aureus - immunology T-Lymphocytes - immunology |
title | An alternative method for T-cell receptor repertoire analysis: Clustering of human V-beta subfamilies selected in responses to staphylococcal enterotoxins B and E |
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