In Vivo Imaging of Transiently Transgenized Mice with a Bovine Interleukin 8 (CXCL8) Promoter/Luciferase Reporter Construct
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{"title"=>"In vivo imaging of transiently transgenized mice with a bovine interleukin 8 (CXCL8) promoter/luciferase reporter construct", "type"=>"journal", "authors"=>[{"first_name"=>"Fabio Franco", "last_name"=>"Stellari", "scopus_author_id"=>"24308628300"}, {"first_name"=>"Valentina", "last_name"=>"Franceschi", "scopus_author_id"=>"25121530000"}, {"first_name"=>"Antonio", "last_name"=>"Capocefalo", "scopus_author_id"=>"25227645400"}, {"first_name"=>"Marcello", "last_name"=>"Ronchei", "scopus_author_id"=>"55266395500"}, {"first_name"=>"Fabrizio", "last_name"=>"Facchinetti", "scopus_author_id"=>"10539014300"}, {"first_name"=>"Gino", "last_name"=>"Villetti", "scopus_author_id"=>"56010796300"}, {"first_name"=>"Gaetano", "last_name"=>"Donofrio", "scopus_author_id"=>"6701753220"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "sgr"=>"84862976029", "doi"=>"10.1371/journal.pone.0039716", "scopus"=>"2-s2.0-84862976029", "pui"=>"365122124", "pmid"=>"22761878"}, "id"=>"5199ddbf-3b69-3766-a09f-fa3a94fe3bc1", "abstract"=>"One of the most remarkable properties of interleukin 8 (CXCL8/IL-8), a chemokine with known additional functions also in angiogenesis and tissue remodeling, is the variation of its expression levels. In healthy tissues, IL-8 is barely detectable, but it is rapidly induced by several folds in response to proinflammatory cytokines, bacterial or viral products, and cellular stress. Although mouse cells do not bear a clear homologous IL-8 gene, the murine transcriptional apparatus may well be capable of activating or repressing a heterologous IL-8 gene promoter driving a reporter gene. In order to induce a transient transgenic expression, mice were systemically injected with a bovine IL-8 promoter–luciferase construct. Subsequently mice were monitored for luciferase expression in the lung by in vivo bioluminescent image analysis over an extended period of time (up to 60 days). We demonstrate that the bovine IL-8 promoter–luciferase construct is transiently and robustly activated 3–5 hours after LPS and TNF-α instillation into the lung, peaking at 35 days after construct delivery. Bovine IL-8 promoter–luciferase activation correlates with white blood cell and neutrophil infiltration into the lung. This study demonstrates that a small experimental rodent model can be utilized for non-invasively monitoring, through a reporter gene system, the activation of an IL-8 promoter region derived from a larger size animal (bovine). This proof of principle study has the potential to be utilized also for studying primate IL-8 promoter regions.", "link"=>"http://www.mendeley.com/research/vivo-imaging-transiently-transgenized-mice-bovine-interleukin-8-cxcl8-promoterluciferase-reporter-co", "reader_count"=>17, "reader_count_by_academic_status"=>{"Unspecified"=>2, "Professor > Associate Professor"=>2, "Researcher"=>2, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>2, "Other"=>2, "Student > Master"=>4, "Student > Bachelor"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>2, "Professor > Associate Professor"=>2, "Researcher"=>2, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>2, "Other"=>2, "Student > Master"=>4, "Student > Bachelor"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>3, "Biochemistry, Genetics and Molecular Biology"=>2, "Agricultural and Biological Sciences"=>9, "Medicine and Dentistry"=>1, "Veterinary Science and Veterinary Medicine"=>1, "Chemistry"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>1}, "Chemistry"=>{"Chemistry"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>9}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>2}, "Unspecified"=>{"Unspecified"=>3}, "Veterinary Science and Veterinary Medicine"=>{"Veterinary Science and Veterinary Medicine"=>1}}, "reader_count_by_country"=>{"Canada"=>1}, "group_count"=>0}

Scopus | Further Information

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Figshare

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  • {"files"=>["https://ndownloader.figshare.com/files/617437"], "description"=>"<p><b>A</b>) Representative images of groups of mice (n = 3 per group) transiently transgenized with bIL-8-Luc or Empty vector DNA and intratracheally instilled with LPS, TNF-α or vehicle (saline solution) at 8, 32, 45 and 60 days post transgenization. Mice were monitored at 3 hours post stimulation by BLI by drawing a region of interest (ROI) over the chest. <b>B</b>) Light intensity quantification of the ROI using the LivingImage software. The experiment was repeated three times and each point represents the mean ± standard deviation of 9 animals. <b>C</b>) Data were expressed as folds of induction over vehicle-treated mice (Saline) and statistical differences were tested by One Way ANOVA followed by Dunnet’s post hoc test for group comparisons. Results are reported as mean ± SD and significance attributed when <i>P</i><0.05 (*) or <i>P</i><0.01(**).</p>", "links"=>[], "tags"=>["lasting", "bil-8-luc"], "article_id"=>287926, "categories"=>["Medicine", "Cell Biology", "Pharmacology", "Immunology", "Physiology", "Biotechnology"], "users"=>["Fabio Franco Stellari", "Valentina Franceschi", "Antonio Capocefalo", "Marcello Ronchei", "Fabrizio Facchinetti", "Gino Villetti", "Gaetano Donofrio"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0039716.g003", "stats"=>{"downloads"=>1, "page_views"=>21, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Long_lasting_response_of_bIL_8_Luc_reporter_construct_/287926", "title"=>"Long lasting response of bIL-8-Luc reporter construct.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-06-28 02:12:06"}
  • {"files"=>["https://ndownloader.figshare.com/files/617538"], "description"=>"<p>Cellular infiltration into the lung of mice intratracheally instilled with LPS, TNF-α or vehicle. The amount of White blood cells (WBC), Neutrophils (Neut) and Monocytes (Mono) found in BALF was expressed as number of cells per μl at 3 (A) and 24 hours (B) post treatment. The experiment was repeated three times and each point represents the mean ± standard deviation of 9 animals. Data were expressed as folds of induction over vehicle-treated mice (Saline) and statistical differences were tested by One Way ANOVA followed by Dunnet’s post hoc test for group comparisons. Results are reported as mean ± SD and significance attributed when <i>P</i><0.05 (*) or <i>P</i><0.01(**).</p>", "links"=>[], "tags"=>["cells", "neutrophils"], "article_id"=>288026, "categories"=>["Medicine", "Cell Biology", "Pharmacology", "Immunology", "Physiology", "Biotechnology"], "users"=>["Fabio Franco Stellari", "Valentina Franceschi", "Antonio Capocefalo", "Marcello Ronchei", "Fabrizio Facchinetti", "Gino Villetti", "Gaetano Donofrio"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0039716.g004", "stats"=>{"downloads"=>1, "page_views"=>14, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_White_blood_cells_and_neutrophils_infiltration_/288026", "title"=>"White blood cells and neutrophils infiltration.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-06-28 02:13:46"}
  • {"files"=>["https://ndownloader.figshare.com/files/321234", "https://ndownloader.figshare.com/files/321305"], "description"=>"<div><p>One of the most remarkable properties of interleukin 8 (CXCL8/IL-8), a chemokine with known additional functions also in angiogenesis and tissue remodeling, is the variation of its expression levels. In healthy tissues, IL-8 is barely detectable, but it is rapidly induced by several folds in response to proinflammatory cytokines, bacterial or viral products, and cellular stress. Although mouse cells do not bear a clear homologous IL-8 gene, the murine transcriptional apparatus may well be capable of activating or repressing a heterologous IL-8 gene promoter driving a reporter gene. In order to induce a transient transgenic expression, mice were systemically injected with a bovine IL-8 promoter–luciferase construct. Subsequently mice were monitored for luciferase expression in the lung by <em>in vivo</em> bioluminescent image analysis over an extended period of time (up to 60 days). We demonstrate that the bovine IL-8 promoter–luciferase construct is transiently and robustly activated 3–5 hours after LPS and TNF-α instillation into the lung, peaking at 35 days after construct delivery. Bovine IL-8 promoter–luciferase activation correlates with white blood cell and neutrophil infiltration into the lung. This study demonstrates that a small experimental rodent model can be utilized for non-invasively monitoring, through a reporter gene system, the activation of an IL-8 promoter region derived from a larger size animal (bovine). This proof of principle study has the potential to be utilized also for studying primate IL-8 promoter regions.</p> </div>", "links"=>[], "tags"=>["imaging", "transiently", "transgenized", "mice", "bovine", "interleukin"], "article_id"=>123402, "categories"=>["Medicine", "Cell Biology", "Pharmacology", "Immunology", "Physiology", "Biotechnology"], "users"=>["Fabio Franco Stellari", "Valentina Franceschi", "Antonio Capocefalo", "Marcello Ronchei", "Fabrizio Facchinetti", "Gino Villetti", "Gaetano Donofrio"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0039716.s001", "https://dx.doi.org/10.1371/journal.pone.0039716.s002"], "stats"=>{"downloads"=>2, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_In_Vivo_Imaging_of_Transiently_Transgenized_Mice_with_a_Bovine_Interleukin_8_CXCL8_Promoter_Luciferase_Reporter_Construct/123402", "title"=>"<em>In Vivo</em> Imaging of Transiently Transgenized Mice with a Bovine Interleukin 8 (CXCL8) Promoter/Luciferase Reporter Construct", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2012-06-28 00:56:42"}
  • {"files"=>["https://ndownloader.figshare.com/files/617161"], "description"=>"<p><b>A</b>) Schematic diagram (not to scale) of the <i>Bos taurus</i> chromosome 6 with the IL-8 gene locus comprising promoter and exons annotated by numbers. The 2 kb IL-8 promoter sequence, comprising the putative TATA box, the transcriptional start site (+1) and the UTR (red underlined) was cloned into the pGL3 basic vector in front of the open reading frame of the Luciferase reporter. <b>B</b>) RT-PCR products of TLRs from 1 to 9 and their corresponding amplicon size. GAPDH was used as an internal control and the amplification performed in the absence of reverse transcriptase (-RT) as a negative control. <b>C</b>) RT-PCR products of TNF-αRI,II and their corresponding amplicon size. GAPDH was used as an internal control and the amplification performed in the absence of reverse transcriptase (-RT) as a negative control. D) IL-8 promoter activation after treatment of bIL-8-Luc transfected LA-4 cells with LPS or TNF-α, along with the untreated control (Untr.). Each experiment was done in quadruplicate, and each point represents the mean ± standard deviation of three experiments. Data were expressed as folds of induction (2,1 and 2,8 for LPS and TNF-α respectively) over vehicle-treated cells and statistical differences were tested by One Way ANOVA followed by Dunnet’s post hoc test for group comparisons. Results are reported as mean ± SD and significance attributed when <i>P</i><0.05 (*) or <i>P</i><0.01(**).</p>", "links"=>[], "tags"=>["characterization", "bil-8-luc"], "article_id"=>287647, "categories"=>["Medicine", "Cell Biology", "Pharmacology", "Immunology", "Physiology", "Biotechnology"], "users"=>["Fabio Franco Stellari", "Valentina Franceschi", "Antonio Capocefalo", "Marcello Ronchei", "Fabrizio Facchinetti", "Gino Villetti", "Gaetano Donofrio"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0039716.g001", "stats"=>{"downloads"=>0, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_In_vitro_characterization_of_bIL_8_Luc_reporter_construct_/287647", "title"=>"<i>In vitro</i> characterization of bIL-8-Luc reporter construct.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-06-28 02:07:27"}

PMC Usage Stats | Further Information

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Relative Metric

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