Determination of haptoglobin expression in IL-6 treated HepG2 cells by ELISA and by RNA hybridization — Evaluation of a quantitative method to measure IL-6
Interleukin-6 (IL-6) is known to be an important modulator of acute phase (AP) protein expression in hepatocytes both in vivo and in vitro. In the present study the inducing activity of IL-6 on the expression of the AP protein haptoglobin (HP) by the human hepatoma cell line HepG2, has been evaluate...
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| Veröffentlicht in: | Journal of immunological methods 1994-05, Vol.171 (2), p.157-167 |
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| creator | Boe, A. Canosi, U. Donini, S. Mastrangeli, R. Ythier, A. Crescenzi, O.Serlupi |
| description | Interleukin-6 (IL-6) is known to be an important modulator of acute phase (AP) protein expression in hepatocytes both in vivo and in vitro. In the present study the inducing activity of IL-6 on the expression of the AP protein haptoglobin (HP) by the human hepatoma cell line HepG2, has been evaluated. HP mRNA inducibility was analysed by Northern and slot-blot hybridization, while HP protein waa detected by means of an ELISA procedure. A dose-response relations from 0.3 to 4.8 ng/ml of a human recombinant IL-6 preparation derived from a Chinese hamster ovary (CHO) cell line was observed after 48 of h treatment. Comparable results were obtained by analysing both HP mRNA expression and HP protein secretion. Detectable induction of HP protein secretion was observed with as little as 25 pg/ml of IL-6. The effect of IL-6 was potentiated by dexamethasone, while an inhibition on HP mRNA inducibility could be prevented by lowering the foetal calf serum (FCS) concentration to 1%. Preliminary data indicate that neither IL-1β nor TNF-α were able to induce significantly HP mRNA expression and protein secretion. The activity ratio between two IL-6 preparations (from CHO and
E. coli cells) obtained with a conventional IL-6 bioassay (i.e., T1165 cell growth assay) was comparable to that obtained in the induction of HP expression. The nominal specific activity of the CHO-derived IL-6 was two to three times higher with both responses. |
| doi_str_mv | 10.1016/0022-1759(94)90036-1 |
| format | Article |
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E. coli cells) obtained with a conventional IL-6 bioassay (i.e., T1165 cell growth assay) was comparable to that obtained in the induction of HP expression. The nominal specific activity of the CHO-derived IL-6 was two to three times higher with both responses.</description><identifier>ISSN: 0022-1759</identifier><identifier>EISSN: 1872-7905</identifier><identifier>DOI: 10.1016/0022-1759(94)90036-1</identifier><identifier>PMID: 8195587</identifier><identifier>CODEN: JIMMBG</identifier><language>eng</language><publisher>Amsterdam: Elsevier B.V</publisher><subject>Acute phase protein ; Analytical, structural and metabolic biochemistry ; Animals ; Base Sequence ; Biological and medical sciences ; Carcinoma, Hepatocellular - genetics ; Carcinoma, Hepatocellular - metabolism ; CHO Cells - drug effects ; Cricetinae ; Dexamethasone ; ELISA ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; Evaluation Studies as Topic ; Filter hybridization ; Fundamental and applied biological sciences. Psychology ; General aspects, investigation methods ; Haptoglobins - biosynthesis ; Haptoglobins - genetics ; Hepatoma cell ; Humans ; IL-6 ; Interleukin-6 - analysis ; Interleukin-6 - pharmacology ; Kinetics ; Molecular Sequence Data ; Nucleic Acid Hybridization ; Proteins ; RNA, Messenger - biosynthesis ; RNA, Messenger - genetics ; Tumor Cells, Cultured - drug effects</subject><ispartof>Journal of immunological methods, 1994-05, Vol.171 (2), p.157-167</ispartof><rights>1994</rights><rights>1994 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c417t-b3f5baa392636a4598f4295931051b57ef3846b69c9890794963300ab90483b43</citedby><cites>FETCH-LOGICAL-c417t-b3f5baa392636a4598f4295931051b57ef3846b69c9890794963300ab90483b43</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/0022175994900361$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65534</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=4066868$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/8195587$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Boe, A.</creatorcontrib><creatorcontrib>Canosi, U.</creatorcontrib><creatorcontrib>Donini, S.</creatorcontrib><creatorcontrib>Mastrangeli, R.</creatorcontrib><creatorcontrib>Ythier, A.</creatorcontrib><creatorcontrib>Crescenzi, O.Serlupi</creatorcontrib><title>Determination of haptoglobin expression in IL-6 treated HepG2 cells by ELISA and by RNA hybridization — Evaluation of a quantitative method to measure IL-6</title><title>Journal of immunological methods</title><addtitle>J Immunol Methods</addtitle><description>Interleukin-6 (IL-6) is known to be an important modulator of acute phase (AP) protein expression in hepatocytes both in vivo and in vitro. In the present study the inducing activity of IL-6 on the expression of the AP protein haptoglobin (HP) by the human hepatoma cell line HepG2, has been evaluated. HP mRNA inducibility was analysed by Northern and slot-blot hybridization, while HP protein waa detected by means of an ELISA procedure. A dose-response relations from 0.3 to 4.8 ng/ml of a human recombinant IL-6 preparation derived from a Chinese hamster ovary (CHO) cell line was observed after 48 of h treatment. Comparable results were obtained by analysing both HP mRNA expression and HP protein secretion. Detectable induction of HP protein secretion was observed with as little as 25 pg/ml of IL-6. The effect of IL-6 was potentiated by dexamethasone, while an inhibition on HP mRNA inducibility could be prevented by lowering the foetal calf serum (FCS) concentration to 1%. Preliminary data indicate that neither IL-1β nor TNF-α were able to induce significantly HP mRNA expression and protein secretion. The activity ratio between two IL-6 preparations (from CHO and
E. coli cells) obtained with a conventional IL-6 bioassay (i.e., T1165 cell growth assay) was comparable to that obtained in the induction of HP expression. The nominal specific activity of the CHO-derived IL-6 was two to three times higher with both responses.</description><subject>Acute phase protein</subject><subject>Analytical, structural and metabolic biochemistry</subject><subject>Animals</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Carcinoma, Hepatocellular - genetics</subject><subject>Carcinoma, Hepatocellular - metabolism</subject><subject>CHO Cells - drug effects</subject><subject>Cricetinae</subject><subject>Dexamethasone</subject><subject>ELISA</subject><subject>Enzyme-Linked Immunosorbent Assay</subject><subject>Escherichia coli</subject><subject>Evaluation Studies as Topic</subject><subject>Filter hybridization</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>General aspects, investigation methods</subject><subject>Haptoglobins - biosynthesis</subject><subject>Haptoglobins - genetics</subject><subject>Hepatoma cell</subject><subject>Humans</subject><subject>IL-6</subject><subject>Interleukin-6 - analysis</subject><subject>Interleukin-6 - pharmacology</subject><subject>Kinetics</subject><subject>Molecular Sequence Data</subject><subject>Nucleic Acid Hybridization</subject><subject>Proteins</subject><subject>RNA, Messenger - biosynthesis</subject><subject>RNA, Messenger - genetics</subject><subject>Tumor Cells, Cultured - drug effects</subject><issn>0022-1759</issn><issn>1872-7905</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1994</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkc2O0zAQxyMEWroLbwCSDwjBITCOP2JfVqqWslupAomPs-UkE2qUr7Wdiu6Jh-DKy_EkJNuqRzjZ4_nNf8bzT5JnFN5QoPItQJalNBf6leavNQCTKX2QLKjKszTXIB4mixPyODkP4TsAUJBwlpwpqoVQ-SL5_Q4j-tZ1Nrq-I31NtnaI_bemL1xH8MfgMYQ5M0XrTSpJ9GgjVuQGh-uMlNg0gRR7stqsPy-J7ao5-PRhSbb7wrvK3R10__z8RVY724ynNpbcjraLLk4vOyQtxm1fkdhPNxtGj_fdniSPatsEfHo8L5Kv71dfrm7Szcfr9dVyk5ac5jEtWC0Ka5nOJJOWC61qnmmhGQVBC5FjzRSXhdSlVhpyzbVkDMAWGrhiBWcXycuD7uD72xFDNK0L899sh_0YTC7FtDih_gtSqRWDDCaQH8DS9yF4rM3gXWv93lAws31m9sbM3hjNzb19hk5lz4_6Y9FidSo6-jXlXxzzNpS2qb3tShdOGAcplZzHvDxgOC1t59CbUDrsSqycxzKaqnf_nuMv4oS1XA</recordid><startdate>19940516</startdate><enddate>19940516</enddate><creator>Boe, A.</creator><creator>Canosi, U.</creator><creator>Donini, S.</creator><creator>Mastrangeli, R.</creator><creator>Ythier, A.</creator><creator>Crescenzi, O.Serlupi</creator><general>Elsevier B.V</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>7QO</scope><scope>7T5</scope><scope>8FD</scope><scope>FR3</scope><scope>H94</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>19940516</creationdate><title>Determination of haptoglobin expression in IL-6 treated HepG2 cells by ELISA and by RNA hybridization — Evaluation of a quantitative method to measure IL-6</title><author>Boe, A. ; Canosi, U. ; Donini, S. ; Mastrangeli, R. ; Ythier, A. ; Crescenzi, O.Serlupi</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c417t-b3f5baa392636a4598f4295931051b57ef3846b69c9890794963300ab90483b43</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1994</creationdate><topic>Acute phase protein</topic><topic>Analytical, structural and metabolic biochemistry</topic><topic>Animals</topic><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>Carcinoma, Hepatocellular - genetics</topic><topic>Carcinoma, Hepatocellular - metabolism</topic><topic>CHO Cells - drug effects</topic><topic>Cricetinae</topic><topic>Dexamethasone</topic><topic>ELISA</topic><topic>Enzyme-Linked Immunosorbent Assay</topic><topic>Escherichia coli</topic><topic>Evaluation Studies as Topic</topic><topic>Filter hybridization</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>General aspects, investigation methods</topic><topic>Haptoglobins - biosynthesis</topic><topic>Haptoglobins - genetics</topic><topic>Hepatoma cell</topic><topic>Humans</topic><topic>IL-6</topic><topic>Interleukin-6 - analysis</topic><topic>Interleukin-6 - pharmacology</topic><topic>Kinetics</topic><topic>Molecular Sequence Data</topic><topic>Nucleic Acid Hybridization</topic><topic>Proteins</topic><topic>RNA, Messenger - biosynthesis</topic><topic>RNA, Messenger - genetics</topic><topic>Tumor Cells, Cultured - drug effects</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Boe, A.</creatorcontrib><creatorcontrib>Canosi, U.</creatorcontrib><creatorcontrib>Donini, S.</creatorcontrib><creatorcontrib>Mastrangeli, R.</creatorcontrib><creatorcontrib>Ythier, A.</creatorcontrib><creatorcontrib>Crescenzi, O.Serlupi</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>Biotechnology Research Abstracts</collection><collection>Immunology Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of immunological methods</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Boe, A.</au><au>Canosi, U.</au><au>Donini, S.</au><au>Mastrangeli, R.</au><au>Ythier, A.</au><au>Crescenzi, O.Serlupi</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Determination of haptoglobin expression in IL-6 treated HepG2 cells by ELISA and by RNA hybridization — Evaluation of a quantitative method to measure IL-6</atitle><jtitle>Journal of immunological methods</jtitle><addtitle>J Immunol Methods</addtitle><date>1994-05-16</date><risdate>1994</risdate><volume>171</volume><issue>2</issue><spage>157</spage><epage>167</epage><pages>157-167</pages><issn>0022-1759</issn><eissn>1872-7905</eissn><coden>JIMMBG</coden><abstract>Interleukin-6 (IL-6) is known to be an important modulator of acute phase (AP) protein expression in hepatocytes both in vivo and in vitro. In the present study the inducing activity of IL-6 on the expression of the AP protein haptoglobin (HP) by the human hepatoma cell line HepG2, has been evaluated. HP mRNA inducibility was analysed by Northern and slot-blot hybridization, while HP protein waa detected by means of an ELISA procedure. A dose-response relations from 0.3 to 4.8 ng/ml of a human recombinant IL-6 preparation derived from a Chinese hamster ovary (CHO) cell line was observed after 48 of h treatment. Comparable results were obtained by analysing both HP mRNA expression and HP protein secretion. Detectable induction of HP protein secretion was observed with as little as 25 pg/ml of IL-6. The effect of IL-6 was potentiated by dexamethasone, while an inhibition on HP mRNA inducibility could be prevented by lowering the foetal calf serum (FCS) concentration to 1%. Preliminary data indicate that neither IL-1β nor TNF-α were able to induce significantly HP mRNA expression and protein secretion. The activity ratio between two IL-6 preparations (from CHO and
E. coli cells) obtained with a conventional IL-6 bioassay (i.e., T1165 cell growth assay) was comparable to that obtained in the induction of HP expression. The nominal specific activity of the CHO-derived IL-6 was two to three times higher with both responses.</abstract><cop>Amsterdam</cop><pub>Elsevier B.V</pub><pmid>8195587</pmid><doi>10.1016/0022-1759(94)90036-1</doi><tpages>11</tpages></addata></record> |
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| ispartof | Journal of immunological methods, 1994-05, Vol.171 (2), p.157-167 |
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| subjects | Acute phase protein Analytical, structural and metabolic biochemistry Animals Base Sequence Biological and medical sciences Carcinoma, Hepatocellular - genetics Carcinoma, Hepatocellular - metabolism CHO Cells - drug effects Cricetinae Dexamethasone ELISA Enzyme-Linked Immunosorbent Assay Escherichia coli Evaluation Studies as Topic Filter hybridization Fundamental and applied biological sciences. Psychology General aspects, investigation methods Haptoglobins - biosynthesis Haptoglobins - genetics Hepatoma cell Humans IL-6 Interleukin-6 - analysis Interleukin-6 - pharmacology Kinetics Molecular Sequence Data Nucleic Acid Hybridization Proteins RNA, Messenger - biosynthesis RNA, Messenger - genetics Tumor Cells, Cultured - drug effects |
| title | Determination of haptoglobin expression in IL-6 treated HepG2 cells by ELISA and by RNA hybridization — Evaluation of a quantitative method to measure IL-6 |
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