Cellular Immunogenicity of Novel Gene Immunogens in Mice Monitored by in Vivo Imaging
The efficient cell-mediated immune response clears cells expressing deoxyribonucleic acid (DNA) immunogens, but there are no methods to monitor this in vivo. We hypothesized that immune-mediated clearance can be monitored in vivo if DNA immunogens are coexpressed with reporter(s). To test this, we d...
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| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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SAGE Publishing
2012-11-01
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| Series: | Molecular Imaging |
| Online Access: | https://doi.org/10.2310/7290.2012.00011 |
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| author | Elizaveta Starodubova Olga Krotova David Hallengärd Yulia Kuzmenko Gunnel Engström Diana Legzdina Oleg Latyshev Olesja Eliseeva Anna Karin Maltais Vera Tunitskaya Vadim Karpov Andreas Bråve Maria Isaguliants |
| author_facet | Elizaveta Starodubova Olga Krotova David Hallengärd Yulia Kuzmenko Gunnel Engström Diana Legzdina Oleg Latyshev Olesja Eliseeva Anna Karin Maltais Vera Tunitskaya Vadim Karpov Andreas Bråve Maria Isaguliants |
| author_sort | Elizaveta Starodubova |
| collection | DOAJ |
| description | The efficient cell-mediated immune response clears cells expressing deoxyribonucleic acid (DNA) immunogens, but there are no methods to monitor this in vivo. We hypothesized that immune-mediated clearance can be monitored in vivo if DNA immunogens are coexpressed with reporter(s). To test this, we designed genes encoding human immunodeficiency virus 1 (HIV-1) reverse transcriptase (RT) fused via its N- or C-terminus to 30–amino acid-long Gly-Ala-repeat of Epstein-Barr virus nuclear antigen 1 or via the N-terminus to the transport signal of invariant chain/Ii or inserted between the cytoplasmic and luminal domains of lysosome-associated membrane protein I (LAMP). DNA immunogens mixed with luciferase gene were injected into BALB/c mice with subsequent electroporation. Reporter expression seen as luminescence was monitored by in vivo imaging. When luminescence faded, mice were sacrificed, and their splenocytes were stimulated with RT-derived antigens. Fading of luminescence correlated with the RT-specific secretion of interferon-γ and interleukin-2. Both immune and in vivo imaging techniques concordantly demonstrated an enhanced immunogenicity of RT-LAMP and of the N-terminal Gly-Ala-RT fusion genes. In vivo imaging performed as an animal-sparing method to estimate the overall performance of DNA immunogens, predicting it early in the experiment. So far, in vivo imaging cannot be a substitute for conventional immune assays, but it is supplementary to them. Further experiments are needed to identify which arms of cellular immune response in vivo imaging monitors best. |
| format | Article |
| id | doaj-art-897be335bc7842948ed120a1e544abf8 |
| institution | Kabale University |
| issn | 1536-0121 |
| language | English |
| publishDate | 2012-11-01 |
| publisher | SAGE Publishing |
| record_format | Article |
| series | Molecular Imaging |
| spelling | doaj-art-897be335bc7842948ed120a1e544abf82025-01-03T00:10:25ZengSAGE PublishingMolecular Imaging1536-01212012-11-011110.2310/7290.2012.0001110.2310_7290.2012.00011Cellular Immunogenicity of Novel Gene Immunogens in Mice Monitored by in Vivo ImagingElizaveta StarodubovaOlga KrotovaDavid HallengärdYulia KuzmenkoGunnel EngströmDiana LegzdinaOleg LatyshevOlesja EliseevaAnna Karin MaltaisVera TunitskayaVadim KarpovAndreas BråveMaria IsaguliantsThe efficient cell-mediated immune response clears cells expressing deoxyribonucleic acid (DNA) immunogens, but there are no methods to monitor this in vivo. We hypothesized that immune-mediated clearance can be monitored in vivo if DNA immunogens are coexpressed with reporter(s). To test this, we designed genes encoding human immunodeficiency virus 1 (HIV-1) reverse transcriptase (RT) fused via its N- or C-terminus to 30–amino acid-long Gly-Ala-repeat of Epstein-Barr virus nuclear antigen 1 or via the N-terminus to the transport signal of invariant chain/Ii or inserted between the cytoplasmic and luminal domains of lysosome-associated membrane protein I (LAMP). DNA immunogens mixed with luciferase gene were injected into BALB/c mice with subsequent electroporation. Reporter expression seen as luminescence was monitored by in vivo imaging. When luminescence faded, mice were sacrificed, and their splenocytes were stimulated with RT-derived antigens. Fading of luminescence correlated with the RT-specific secretion of interferon-γ and interleukin-2. Both immune and in vivo imaging techniques concordantly demonstrated an enhanced immunogenicity of RT-LAMP and of the N-terminal Gly-Ala-RT fusion genes. In vivo imaging performed as an animal-sparing method to estimate the overall performance of DNA immunogens, predicting it early in the experiment. So far, in vivo imaging cannot be a substitute for conventional immune assays, but it is supplementary to them. Further experiments are needed to identify which arms of cellular immune response in vivo imaging monitors best.https://doi.org/10.2310/7290.2012.00011 |
| spellingShingle | Elizaveta Starodubova Olga Krotova David Hallengärd Yulia Kuzmenko Gunnel Engström Diana Legzdina Oleg Latyshev Olesja Eliseeva Anna Karin Maltais Vera Tunitskaya Vadim Karpov Andreas Bråve Maria Isaguliants Cellular Immunogenicity of Novel Gene Immunogens in Mice Monitored by in Vivo Imaging Molecular Imaging |
| title | Cellular Immunogenicity of Novel Gene Immunogens in Mice Monitored by in Vivo Imaging |
| title_full | Cellular Immunogenicity of Novel Gene Immunogens in Mice Monitored by in Vivo Imaging |
| title_fullStr | Cellular Immunogenicity of Novel Gene Immunogens in Mice Monitored by in Vivo Imaging |
| title_full_unstemmed | Cellular Immunogenicity of Novel Gene Immunogens in Mice Monitored by in Vivo Imaging |
| title_short | Cellular Immunogenicity of Novel Gene Immunogens in Mice Monitored by in Vivo Imaging |
| title_sort | cellular immunogenicity of novel gene immunogens in mice monitored by in vivo imaging |
| url | https://doi.org/10.2310/7290.2012.00011 |
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