Direct Assessment of Cellular Immune Responses to Ovalbumin with a Secreted Luciferase Transgenic Reporter Mouse Strain IFN纬-Lucia

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Immune Responses to Ovalbumin with a Secreted Luciferase Transgenic Reporter Mouse Strain IFN纬-Lucia</title> <meta name="description" content="Direct Assessment of Cellular Immune Responses to Ovalbumin with a Secreted Luciferase Transgenic Reporter Mouse Strain IFN纬-Lucia"> <meta name="keywords" content="ELISpot, immunogenicity, interferon-gamma, reporter mice, vaccines"> <meta name="viewport" content="width=device-width, initial-scale=1, minimum-scale=1, maximum-scale=1, user-scalable=no"> <meta charset="utf-8"> <link href="https://cdn.waset.org/favicon.ico" type="image/x-icon" rel="shortcut icon"> <link href="https://cdn.waset.org/static/plugins/bootstrap-4.2.1/css/bootstrap.min.css" rel="stylesheet"> <link href="https://cdn.waset.org/static/plugins/fontawesome/css/all.min.css" rel="stylesheet"> <link href="https://cdn.waset.org/static/css/site.css?v=150220211555" rel="stylesheet"> </head> <body> <header> <div class="container"> <nav class="navbar navbar-expand-lg navbar-light"> <a 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87356</div> </div> </div> </div> <div class="card publication-listing mt-3 mb-3"> <h5 class="card-header" style="font-size:.9rem">Direct Assessment of Cellular Immune Responses to Ovalbumin with a Secreted Luciferase Transgenic Reporter Mouse Strain IFN纬-Lucia</h5> <div class="card-body"> <p class="card-text"><strong>Authors:</strong> <a href="https://publications.waset.org/abstracts/search?q=Martyna%20Chotomska">Martyna Chotomska</a>, <a href="https://publications.waset.org/abstracts/search?q=Aleksandra%20Studzinska"> Aleksandra Studzinska</a>, <a href="https://publications.waset.org/abstracts/search?q=Marta%20Lisowska"> Marta Lisowska</a>, <a href="https://publications.waset.org/abstracts/search?q=Justyna%20Szubert"> Justyna Szubert</a>, <a href="https://publications.waset.org/abstracts/search?q=Aleksandra%20Tabis"> Aleksandra Tabis</a>, <a href="https://publications.waset.org/abstracts/search?q=Jacek%20Bania"> Jacek Bania</a>, <a href="https://publications.waset.org/abstracts/search?q=Arkadiusz%20Miazek"> Arkadiusz Miazek</a> </p> <p class="card-text"><strong>Abstract:</strong></p> Objectives: Assessing antigen-specific T cell responses is of utmost importance for the pre-clinical testing of prototype vaccines against intracellular pathogens and tumor antigens. Mainly two types of in vitro assays are used for this purpose 1) enzyme-linked immunospot (ELISpot) and 2) intracellular cytokine staining (ICS). Both are time-consuming, relatively expensive, and require manual dexterity. Here, we assess if a straightforward detection of luciferase activity in blood samples of transgenic reporter mice expressing a secreted Lucia luciferase under the transcriptional control of IFN-纬 promoter parallels the sensitivity of IFN纬 ELISpot assay. Methods: IFN-纬-LUCIA mouse strain carrying multiple copies of Lucia luciferase transgene under the transcriptional control of IFN纬 minimal promoter were generated by pronuclear injection of linear DNA. The specificity of transgene expression and mobilization was assessed in vitro using transgenic splenocytes exposed to various mitogens. The IFN-纬-LUCIA mice were immunized with 50mg of ovalbumin (OVA) emulsified in incomplete Freund鈥檚 adjuvant three times every two weeks by subcutaneous injections. Blood samples were collected before and five days after each immunization. Luciferase activity was assessed in blood serum. Peripheral blood mononuclear cells were separated and assessed for frequencies of OVA-specific IFN纬-secreting T cells. Results: We show that in vitro cultured splenocytes of IFN-纬-LUCIA mice respond by 2 and 3 fold increase in secreted luciferase activity to T cell mitogens concanavalin A and phorbol myristate acetate, respectively but fail to respond to B cell-stimulating E.coli lipopolysaccharide. Immunization of IFN-纬-LUCIA mice with OVA leads to over 4 fold increase in luciferase activity in blood serum five days post-immunization with a barely detectable increase in OVA-specific, IFN纬-secreting T cells by ELISpot. Second and third immunizations, further increase the luciferase activity and coincidently also increase the frequencies of OVA-specific T cells by ELISpot. Conclusions: We conclude that minimally invasive monitoring of luciferase secretions in blood serum of IFN-纬-LUCIA mice constitutes a sensitive method for evaluating primary and memory Th1 responses to protein antigens. As such, this method may complement existing methods for rapid immunogenicity assessment of prototype vaccines. <iframe src="https://publications.waset.org/abstracts/134252.pdf" style="width:100%; height:400px;" frameborder="0"></iframe> <p class="card-text"><strong>Keywords:</strong> <a href="https://publications.waset.org/abstracts/search?q=ELISpot" title="ELISpot">ELISpot</a>, <a href="https://publications.waset.org/abstracts/search?q=immunogenicity" title=" immunogenicity"> immunogenicity</a>, <a href="https://publications.waset.org/abstracts/search?q=interferon-gamma" title=" interferon-gamma"> interferon-gamma</a>, <a href="https://publications.waset.org/abstracts/search?q=reporter%20mice" title=" reporter mice"> reporter mice</a>, <a href="https://publications.waset.org/abstracts/search?q=vaccines" title=" vaccines"> vaccines</a> </p> <a href="https://publications.waset.org/abstracts/134252/direct-assessment-of-cellular-immune-responses-to-ovalbumin-with-a-secreted-luciferase-transgenic-reporter-mouse-strain-ifngh-lucia" class="btn btn-primary btn-sm">Procedia</a> <a href="https://publications.waset.org/abstracts/134252.pdf" target="_blank" class="btn btn-primary btn-sm">PDF</a> <span class="bg-info text-light px-1 py-1 float-right rounded"> Downloads <span class="badge badge-light">170</span> </span> </div> </div> </div> </main> <footer> <div id="infolinks" class="pt-3 pb-2"> <div class="container"> <div style="background-color:#f5f5f5;" class="p-3"> <div class="row"> <div class="col-md-2"> <ul class="list-unstyled"> About <li><a href="https://waset.org/page/support">About Us</a></li> <li><a href="https://waset.org/page/support#legal-information">Legal</a></li> <li><a target="_blank" rel="nofollow" href="https://publications.waset.org/static/files/WASET-16th-foundational-anniversary.pdf">WASET celebrates its 16th 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Direct Assessment of Cellular Immune Responses to Ovalbumin with a Secreted Luciferase Transgenic Reporter Mouse Strain IFN纬-Lucia