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Immunization of BALB/c mice with TERT DNA, TERT variant with a C-terminal hemagglutinin tag (TERT-HA DNA), and empty vector followed by booster immunization with given plasmids mixed with DNA encoding firefly luciferase (Luc DNA), with follow up of luciferase expression by in vivo imaging. Scheme of the immunization ( A ); results of in vivo bioluminescence imaging of booster sites at days 1–12 post injection, example of 3 mice—one from TERT, one from TERT-HA, and one from empty vector group ( B ); dynamics of bioluminescence signal change in TERT, TERT-HA DNA, and empty vector-immunized mice on days 1–12 post booster injection; each line of different colors corresponds to one site of injection (two per mouse) ( C ); relative average level of bioluminescence signal for each group on days 1 to 12 post booster injection ( D ). Bioluminescence signal is represented by the total flux from site of immunization, mean ± SD. Analyzed by ordinary two-way ANOVA with <t>Dunnett’s</t> multiple comparison test, ** − p < 0.01; **** − p < 0.0001; ns—not significant.
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Immunization of BALB/c mice with TERT DNA, TERT variant with a C-terminal hemagglutinin tag (TERT-HA DNA), and empty vector followed by booster immunization with given plasmids mixed with DNA encoding firefly luciferase (Luc DNA), with follow up of luciferase expression by in vivo imaging. Scheme of the immunization ( A ); results of in vivo bioluminescence imaging of booster sites at days 1–12 post injection, example of 3 mice—one from TERT, one from TERT-HA, and one from empty vector group ( B ); dynamics of bioluminescence signal change in TERT, TERT-HA DNA, and empty vector-immunized mice on days 1–12 post booster injection; each line of different colors corresponds to one site of injection (two per mouse) ( C ); relative average level of bioluminescence signal for each group on days 1 to 12 post booster injection ( D ). Bioluminescence signal is represented by the total flux from site of immunization, mean ± SD. Analyzed by ordinary two-way ANOVA with <t>Dunnett’s</t> multiple comparison test, ** − p < 0.01; **** − p < 0.0001; ns—not significant.
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Immunization of BALB/c mice with TERT DNA, TERT variant with a C-terminal hemagglutinin tag (TERT-HA DNA), and empty vector followed by booster immunization with given plasmids mixed with DNA encoding firefly luciferase (Luc DNA), with follow up of luciferase expression by in vivo imaging. Scheme of the immunization ( A ); results of in vivo bioluminescence imaging of booster sites at days 1–12 post injection, example of 3 mice—one from TERT, one from TERT-HA, and one from empty vector group ( B ); dynamics of bioluminescence signal change in TERT, TERT-HA DNA, and empty vector-immunized mice on days 1–12 post booster injection; each line of different colors corresponds to one site of injection (two per mouse) ( C ); relative average level of bioluminescence signal for each group on days 1 to 12 post booster injection ( D ). Bioluminescence signal is represented by the total flux from site of immunization, mean ± SD. Analyzed by ordinary two-way ANOVA with <t>Dunnett’s</t> multiple comparison test, ** − p < 0.01; **** − p < 0.0001; ns—not significant.
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Immunization of BALB/c mice with TERT DNA, TERT variant with a C-terminal hemagglutinin tag (TERT-HA DNA), and empty vector followed by booster immunization with given plasmids mixed with DNA encoding firefly luciferase (Luc DNA), with follow up of luciferase expression by in vivo imaging. Scheme of the immunization ( A ); results of in vivo bioluminescence imaging of booster sites at days 1–12 post injection, example of 3 mice—one from TERT, one from TERT-HA, and one from empty vector group ( B ); dynamics of bioluminescence signal change in TERT, TERT-HA DNA, and empty vector-immunized mice on days 1–12 post booster injection; each line of different colors corresponds to one site of injection (two per mouse) ( C ); relative average level of bioluminescence signal for each group on days 1 to 12 post booster injection ( D ). Bioluminescence signal is represented by the total flux from site of immunization, mean ± SD. Analyzed by ordinary two-way ANOVA with <t>Dunnett’s</t> multiple comparison test, ** − p < 0.01; **** − p < 0.0001; ns—not significant.
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Immunization of BALB/c mice with TERT DNA, TERT variant with a C-terminal hemagglutinin tag (TERT-HA DNA), and empty vector followed by booster immunization with given plasmids mixed with DNA encoding firefly luciferase (Luc DNA), with follow up of luciferase expression by in vivo imaging. Scheme of the immunization ( A ); results of in vivo bioluminescence imaging of booster sites at days 1–12 post injection, example of 3 mice—one from TERT, one from TERT-HA, and one from empty vector group ( B ); dynamics of bioluminescence signal change in TERT, TERT-HA DNA, and empty vector-immunized mice on days 1–12 post booster injection; each line of different colors corresponds to one site of injection (two per mouse) ( C ); relative average level of bioluminescence signal for each group on days 1 to 12 post booster injection ( D ). Bioluminescence signal is represented by the total flux from site of immunization, mean ± SD. Analyzed by ordinary two-way ANOVA with <t>Dunnett’s</t> multiple comparison test, ** − p < 0.01; **** − p < 0.0001; ns—not significant.
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Immunization of BALB/c mice with TERT DNA, TERT variant with a C-terminal hemagglutinin tag (TERT-HA DNA), and empty vector followed by booster immunization with given plasmids mixed with DNA encoding firefly luciferase (Luc DNA), with follow up of luciferase expression by in vivo imaging. Scheme of the immunization ( A ); results of in vivo bioluminescence imaging of booster sites at days 1–12 post injection, example of 3 mice—one from TERT, one from TERT-HA, and one from empty vector group ( B ); dynamics of bioluminescence signal change in TERT, TERT-HA DNA, and empty vector-immunized mice on days 1–12 post booster injection; each line of different colors corresponds to one site of injection (two per mouse) ( C ); relative average level of bioluminescence signal for each group on days 1 to 12 post booster injection ( D ). Bioluminescence signal is represented by the total flux from site of immunization, mean ± SD. Analyzed by ordinary two-way ANOVA with <t>Dunnett’s</t> multiple comparison test, ** − p < 0.01; **** − p < 0.0001; ns—not significant.
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Immunization of BALB/c mice with TERT DNA, TERT variant with a C-terminal hemagglutinin tag (TERT-HA DNA), and empty vector followed by booster immunization with given plasmids mixed with DNA encoding firefly luciferase (Luc DNA), with follow up of luciferase expression by in vivo imaging. Scheme of the immunization ( A ); results of in vivo bioluminescence imaging of booster sites at days 1–12 post injection, example of 3 mice—one from TERT, one from TERT-HA, and one from empty vector group ( B ); dynamics of bioluminescence signal change in TERT, TERT-HA DNA, and empty vector-immunized mice on days 1–12 post booster injection; each line of different colors corresponds to one site of injection (two per mouse) ( C ); relative average level of bioluminescence signal for each group on days 1 to 12 post booster injection ( D ). Bioluminescence signal is represented by the total flux from site of immunization, mean ± SD. Analyzed by ordinary two-way ANOVA with <t>Dunnett’s</t> multiple comparison test, ** − p < 0.01; **** − p < 0.0001; ns—not significant.
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Immunization of BALB/c mice with TERT DNA, TERT variant with a C-terminal hemagglutinin tag (TERT-HA DNA), and empty vector followed by booster immunization with given plasmids mixed with DNA encoding firefly luciferase (Luc DNA), with follow up of luciferase expression by in vivo imaging. Scheme of the immunization ( A ); results of in vivo bioluminescence imaging of booster sites at days 1–12 post injection, example of 3 mice—one from TERT, one from TERT-HA, and one from empty vector group ( B ); dynamics of bioluminescence signal change in TERT, TERT-HA DNA, and empty vector-immunized mice on days 1–12 post booster injection; each line of different colors corresponds to one site of injection (two per mouse) ( C ); relative average level of bioluminescence signal for each group on days 1 to 12 post booster injection ( D ). Bioluminescence signal is represented by the total flux from site of immunization, mean ± SD. Analyzed by ordinary two-way ANOVA with <t>Dunnett’s</t> multiple comparison test, ** − p < 0.01; **** − p < 0.0001; ns—not significant.
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Immunization of BALB/c mice with TERT DNA, TERT variant with a C-terminal hemagglutinin tag (TERT-HA DNA), and empty vector followed by booster immunization with given plasmids mixed with DNA encoding firefly luciferase (Luc DNA), with follow up of luciferase expression by in vivo imaging. Scheme of the immunization ( A ); results of in vivo bioluminescence imaging of booster sites at days 1–12 post injection, example of 3 mice—one from TERT, one from TERT-HA, and one from empty vector group ( B ); dynamics of bioluminescence signal change in TERT, TERT-HA DNA, and empty vector-immunized mice on days 1–12 post booster injection; each line of different colors corresponds to one site of injection (two per mouse) ( C ); relative average level of bioluminescence signal for each group on days 1 to 12 post booster injection ( D ). Bioluminescence signal is represented by the total flux from site of immunization, mean ± SD. Analyzed by ordinary two-way ANOVA with Dunnett’s multiple comparison test, ** − p < 0.01; **** − p < 0.0001; ns—not significant.

Journal: Vaccines

Article Title: Expression of the Reverse Transcriptase Domain of Telomerase Reverse Transcriptase Induces Lytic Cellular Response in DNA-Immunized Mice and Limits Tumorigenic and Metastatic Potential of Murine Adenocarcinoma 4T1 Cells

doi: 10.3390/vaccines8020318

Figure Lengend Snippet: Immunization of BALB/c mice with TERT DNA, TERT variant with a C-terminal hemagglutinin tag (TERT-HA DNA), and empty vector followed by booster immunization with given plasmids mixed with DNA encoding firefly luciferase (Luc DNA), with follow up of luciferase expression by in vivo imaging. Scheme of the immunization ( A ); results of in vivo bioluminescence imaging of booster sites at days 1–12 post injection, example of 3 mice—one from TERT, one from TERT-HA, and one from empty vector group ( B ); dynamics of bioluminescence signal change in TERT, TERT-HA DNA, and empty vector-immunized mice on days 1–12 post booster injection; each line of different colors corresponds to one site of injection (two per mouse) ( C ); relative average level of bioluminescence signal for each group on days 1 to 12 post booster injection ( D ). Bioluminescence signal is represented by the total flux from site of immunization, mean ± SD. Analyzed by ordinary two-way ANOVA with Dunnett’s multiple comparison test, ** − p < 0.01; **** − p < 0.0001; ns—not significant.

Article Snippet: Total photon flux from the site of coinjection of TERT DNA or TERT-HA DNA and Luc DNA was compared using ordinary two-way ANOVA with Dunnett’s multiple comparison test (GraphPad Prism 6, San Diego, CA, USA).

Techniques: Variant Assay, Plasmid Preparation, Luciferase, Expressing, In Vivo Imaging, In Vivo, Imaging, Injection, Comparison

Generation of tumors by 4T1luc2 clones expressing rtTERT. Tumor growth rate was assessed using total fluorescence signal from the site of injection of 2500 ( A ), 5000 ( B ), and 10,000 ( C ) cells. Tumor volume was evaluated by total fluorescence signal from the site of cell injection by day 16 ( D ) or by calipering at day 21 ( E ). Histochemical characterization of the solid tumors formed by the parental 4T1luc2 cells ( F ) and their derivatives expressing rtTERT 4T1luc2_rtTERT_C6 ( G ); 4T1luc2_rtTERT_H9 ( H ) after ectopic implantation into BALB/c mice (H&E staining, magnification 200×). Results of tumor growth ( A – C ) were analyzed using RM two-way ANOVA with Dunnett’s multiple comparison test. Data on tumor volumes ( D , E ) were analyzed using Kruskal–Wallis with Dunn’s multiple comparison test. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001; ns—not significant.

Journal: Vaccines

Article Title: Expression of the Reverse Transcriptase Domain of Telomerase Reverse Transcriptase Induces Lytic Cellular Response in DNA-Immunized Mice and Limits Tumorigenic and Metastatic Potential of Murine Adenocarcinoma 4T1 Cells

doi: 10.3390/vaccines8020318

Figure Lengend Snippet: Generation of tumors by 4T1luc2 clones expressing rtTERT. Tumor growth rate was assessed using total fluorescence signal from the site of injection of 2500 ( A ), 5000 ( B ), and 10,000 ( C ) cells. Tumor volume was evaluated by total fluorescence signal from the site of cell injection by day 16 ( D ) or by calipering at day 21 ( E ). Histochemical characterization of the solid tumors formed by the parental 4T1luc2 cells ( F ) and their derivatives expressing rtTERT 4T1luc2_rtTERT_C6 ( G ); 4T1luc2_rtTERT_H9 ( H ) after ectopic implantation into BALB/c mice (H&E staining, magnification 200×). Results of tumor growth ( A – C ) were analyzed using RM two-way ANOVA with Dunnett’s multiple comparison test. Data on tumor volumes ( D , E ) were analyzed using Kruskal–Wallis with Dunn’s multiple comparison test. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001; ns—not significant.

Article Snippet: Total photon flux from the site of coinjection of TERT DNA or TERT-HA DNA and Luc DNA was compared using ordinary two-way ANOVA with Dunnett’s multiple comparison test (GraphPad Prism 6, San Diego, CA, USA).

Techniques: Clone Assay, Expressing, Fluorescence, Injection, Staining, Comparison