Validation of an Mx/CAT reporter gene assay for the quantification of bovine type-I interferon
We describe here a specific and sensitive assay for biologically active bovine type-I interferon (IFN) in an Mx/CAT reporter gene assay. The assay is based on Madin–Darby Bovine Kidney cells transfected with a plasmid, containing a human MxA promoter driving a chloramphenicol acetyltransferase (CAT)...
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| Veröffentlicht in: | Journal of immunological methods 2001-03, Vol.249 (1), p.235-244 |
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| creator | Fray, M.D. Mann, G.E Charleston, B. |
| description | We describe here a specific and sensitive assay for biologically active bovine type-I interferon (IFN) in an
Mx/CAT reporter gene assay. The assay is based on Madin–Darby Bovine Kidney cells transfected with a plasmid, containing a human
MxA promoter driving a chloramphenicol acetyltransferase (CAT) cDNA. CAT expression was quantified in a commercially available enzyme linked immunosorbant assay. The response to recombinant bovine INF-α
1 was dose dependent between 0.25 and 125.0 iu/ml and was shown to be specific for type-I IFN as no significant effect was seen with a number of other cytokines, including IFN-γ. This
Mx/CAT reporter assay also has advantages in terms of simplicity and reliability over conventional cytopathic effect reduction assays used to quantify the IFN activity in bovine samples. The
Mx/CAT reporter assay was used successfully to measure trophoblast derived type-1 IFN activity (IFN-τ) in uterine flushings collected from pregnant cows. IFN-τ is the pregnancy recognition signal produced in ruminants by pre-implantation embryos and was shown to increase markedly between the 12th (0.7±0.14 iu/ml) and 18th (44 085.0±14 414.2 iu/ml) day of pregnancy. In contrast, IFN-τ activity remained basal (0.5–0.7 iu/ml) in inseminated non-pregnant animals. Duplicate samples analysed using a cytopathic effect reduction assay correlated well (
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| doi_str_mv | 10.1016/S0022-1759(00)00359-8 |
| format | Article |
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Mx/CAT reporter gene assay. The assay is based on Madin–Darby Bovine Kidney cells transfected with a plasmid, containing a human
MxA promoter driving a chloramphenicol acetyltransferase (CAT) cDNA. CAT expression was quantified in a commercially available enzyme linked immunosorbant assay. The response to recombinant bovine INF-α
1 was dose dependent between 0.25 and 125.0 iu/ml and was shown to be specific for type-I IFN as no significant effect was seen with a number of other cytokines, including IFN-γ. This
Mx/CAT reporter assay also has advantages in terms of simplicity and reliability over conventional cytopathic effect reduction assays used to quantify the IFN activity in bovine samples. The
Mx/CAT reporter assay was used successfully to measure trophoblast derived type-1 IFN activity (IFN-τ) in uterine flushings collected from pregnant cows. IFN-τ is the pregnancy recognition signal produced in ruminants by pre-implantation embryos and was shown to increase markedly between the 12th (0.7±0.14 iu/ml) and 18th (44 085.0±14 414.2 iu/ml) day of pregnancy. In contrast, IFN-τ activity remained basal (0.5–0.7 iu/ml) in inseminated non-pregnant animals. Duplicate samples analysed using a cytopathic effect reduction assay correlated well (
P<0.001;
r
2=0.945) with IFN levels obtained using the
Mx/CAT reporter assay, confirming the reporter assay as a reliable substitute for the standard anti-viral IFN assay.</description><identifier>ISSN: 0022-1759</identifier><identifier>EISSN: 1872-7905</identifier><identifier>DOI: 10.1016/S0022-1759(00)00359-8</identifier><identifier>PMID: 11226480</identifier><identifier>CODEN: JIMMBG</identifier><language>eng</language><publisher>Amsterdam: Elsevier B.V</publisher><subject>Animal viral diseases ; Animals ; Bacteriology ; Biological and medical sciences ; Biological Assay ; Cattle ; Chloramphenicol O-Acetyltransferase ; Female ; Fundamental and applied biological sciences. Psychology ; Genes, Reporter ; Genetics ; Humans ; Infectious diseases ; Interferon Type I - analysis ; Interferon Type I - genetics ; MDBK cells ; Medical sciences ; Microbiology ; Pregnancy ; Recombinant Proteins ; Reporter gene assay ; t-Interferon ; Type-I interferon ; Viral diseases</subject><ispartof>Journal of immunological methods, 2001-03, Vol.249 (1), p.235-244</ispartof><rights>2001 Elsevier Science B.V.</rights><rights>2002 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c540t-11d9d1ca0e070662736641e1a38d3d01fa7ec720f707f3e37e7254bf73be82233</citedby><cites>FETCH-LOGICAL-c540t-11d9d1ca0e070662736641e1a38d3d01fa7ec720f707f3e37e7254bf73be82233</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0022175900003598$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3536,27903,27904,65309</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=14164092$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/11226480$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Fray, M.D.</creatorcontrib><creatorcontrib>Mann, G.E</creatorcontrib><creatorcontrib>Charleston, B.</creatorcontrib><title>Validation of an Mx/CAT reporter gene assay for the quantification of bovine type-I interferon</title><title>Journal of immunological methods</title><addtitle>J Immunol Methods</addtitle><description>We describe here a specific and sensitive assay for biologically active bovine type-I interferon (IFN) in an
Mx/CAT reporter gene assay. The assay is based on Madin–Darby Bovine Kidney cells transfected with a plasmid, containing a human
MxA promoter driving a chloramphenicol acetyltransferase (CAT) cDNA. CAT expression was quantified in a commercially available enzyme linked immunosorbant assay. The response to recombinant bovine INF-α
1 was dose dependent between 0.25 and 125.0 iu/ml and was shown to be specific for type-I IFN as no significant effect was seen with a number of other cytokines, including IFN-γ. This
Mx/CAT reporter assay also has advantages in terms of simplicity and reliability over conventional cytopathic effect reduction assays used to quantify the IFN activity in bovine samples. The
Mx/CAT reporter assay was used successfully to measure trophoblast derived type-1 IFN activity (IFN-τ) in uterine flushings collected from pregnant cows. IFN-τ is the pregnancy recognition signal produced in ruminants by pre-implantation embryos and was shown to increase markedly between the 12th (0.7±0.14 iu/ml) and 18th (44 085.0±14 414.2 iu/ml) day of pregnancy. In contrast, IFN-τ activity remained basal (0.5–0.7 iu/ml) in inseminated non-pregnant animals. Duplicate samples analysed using a cytopathic effect reduction assay correlated well (
P<0.001;
r
2=0.945) with IFN levels obtained using the
Mx/CAT reporter assay, confirming the reporter assay as a reliable substitute for the standard anti-viral IFN assay.</description><subject>Animal viral diseases</subject><subject>Animals</subject><subject>Bacteriology</subject><subject>Biological and medical sciences</subject><subject>Biological Assay</subject><subject>Cattle</subject><subject>Chloramphenicol O-Acetyltransferase</subject><subject>Female</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Genes, Reporter</subject><subject>Genetics</subject><subject>Humans</subject><subject>Infectious diseases</subject><subject>Interferon Type I - analysis</subject><subject>Interferon Type I - genetics</subject><subject>MDBK cells</subject><subject>Medical sciences</subject><subject>Microbiology</subject><subject>Pregnancy</subject><subject>Recombinant Proteins</subject><subject>Reporter gene assay</subject><subject>t-Interferon</subject><subject>Type-I interferon</subject><subject>Viral diseases</subject><issn>0022-1759</issn><issn>1872-7905</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2001</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqF0cFuEzEQBmALgWhaeASQLyA4LJ2xd-3dE6oiCpWKOFA4YjneMRht7NTeVOTtcZuoPfbky_fPWP8w9grhAwKq0-8AQjSou-EdwHsA2Q1N_4QtsNei0QN0T9ninhyx41L-AgCCgufsCFEI1fawYL9-2imMdg4p8uS5jfzrv9Pl2RXPtEl5psx_UyRuS7E77lPm8x_i11sb5-CDu8-t0k2obN5tqLngIdagp5ziC_bM26nQy8N7wn6cf7pafmkuv32-WJ5dNq5rYW4Qx2FEZ4FAg1JCS6VaJLSyH-UI6K0mpwV4DdpLkpq06NqV13JFvRBSnrC3-7mbnK63VGazDsXRNNlIaVtMndqBGNSjEHUvqxsq7PbQ5VRKJm82Oaxt3hkEc3sBc3cBc1uvATB3FzB9zb0-LNiu1jQ-pA6VV_DmAGxxdvLZRhfKg2tRtTCI6j7uHdXebgJlU1yg6GgMmdxsxhQe-cp_VtehLg</recordid><startdate>20010301</startdate><enddate>20010301</enddate><creator>Fray, M.D.</creator><creator>Mann, G.E</creator><creator>Charleston, B.</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>20010301</creationdate><title>Validation of an Mx/CAT reporter gene assay for the quantification of bovine type-I interferon</title><author>Fray, M.D. ; Mann, G.E ; Charleston, B.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c540t-11d9d1ca0e070662736641e1a38d3d01fa7ec720f707f3e37e7254bf73be82233</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2001</creationdate><topic>Animal viral diseases</topic><topic>Animals</topic><topic>Bacteriology</topic><topic>Biological and medical sciences</topic><topic>Biological Assay</topic><topic>Cattle</topic><topic>Chloramphenicol O-Acetyltransferase</topic><topic>Female</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Genes, Reporter</topic><topic>Genetics</topic><topic>Humans</topic><topic>Infectious diseases</topic><topic>Interferon Type I - analysis</topic><topic>Interferon Type I - genetics</topic><topic>MDBK cells</topic><topic>Medical sciences</topic><topic>Microbiology</topic><topic>Pregnancy</topic><topic>Recombinant Proteins</topic><topic>Reporter gene assay</topic><topic>t-Interferon</topic><topic>Type-I interferon</topic><topic>Viral diseases</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Fray, M.D.</creatorcontrib><creatorcontrib>Mann, G.E</creatorcontrib><creatorcontrib>Charleston, B.</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>Fray, M.D.</au><au>Mann, G.E</au><au>Charleston, B.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Validation of an Mx/CAT reporter gene assay for the quantification of bovine type-I interferon</atitle><jtitle>Journal of immunological methods</jtitle><addtitle>J Immunol Methods</addtitle><date>2001-03-01</date><risdate>2001</risdate><volume>249</volume><issue>1</issue><spage>235</spage><epage>244</epage><pages>235-244</pages><issn>0022-1759</issn><eissn>1872-7905</eissn><coden>JIMMBG</coden><abstract>We describe here a specific and sensitive assay for biologically active bovine type-I interferon (IFN) in an
Mx/CAT reporter gene assay. The assay is based on Madin–Darby Bovine Kidney cells transfected with a plasmid, containing a human
MxA promoter driving a chloramphenicol acetyltransferase (CAT) cDNA. CAT expression was quantified in a commercially available enzyme linked immunosorbant assay. The response to recombinant bovine INF-α
1 was dose dependent between 0.25 and 125.0 iu/ml and was shown to be specific for type-I IFN as no significant effect was seen with a number of other cytokines, including IFN-γ. This
Mx/CAT reporter assay also has advantages in terms of simplicity and reliability over conventional cytopathic effect reduction assays used to quantify the IFN activity in bovine samples. The
Mx/CAT reporter assay was used successfully to measure trophoblast derived type-1 IFN activity (IFN-τ) in uterine flushings collected from pregnant cows. IFN-τ is the pregnancy recognition signal produced in ruminants by pre-implantation embryos and was shown to increase markedly between the 12th (0.7±0.14 iu/ml) and 18th (44 085.0±14 414.2 iu/ml) day of pregnancy. In contrast, IFN-τ activity remained basal (0.5–0.7 iu/ml) in inseminated non-pregnant animals. Duplicate samples analysed using a cytopathic effect reduction assay correlated well (
P<0.001;
r
2=0.945) with IFN levels obtained using the
Mx/CAT reporter assay, confirming the reporter assay as a reliable substitute for the standard anti-viral IFN assay.</abstract><cop>Amsterdam</cop><pub>Elsevier B.V</pub><pmid>11226480</pmid><doi>10.1016/S0022-1759(00)00359-8</doi><tpages>10</tpages></addata></record> |
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| subjects | Animal viral diseases Animals Bacteriology Biological and medical sciences Biological Assay Cattle Chloramphenicol O-Acetyltransferase Female Fundamental and applied biological sciences. Psychology Genes, Reporter Genetics Humans Infectious diseases Interferon Type I - analysis Interferon Type I - genetics MDBK cells Medical sciences Microbiology Pregnancy Recombinant Proteins Reporter gene assay t-Interferon Type-I interferon Viral diseases |
| title | Validation of an Mx/CAT reporter gene assay for the quantification of bovine type-I interferon |
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