tbev ns1 Search Results


recombinant ns1 protein  (Native Antigen Inc)
94
Native Antigen Inc recombinant ns1 protein
Recombinant Ns1 Protein, supplied by Native Antigen Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/tbev+ns1/Tick-Borne+Encephalitis+Virus+(European)NS1+Protein/pmc07098735-121-0-6
Average 94 stars, based on 1 article reviews
recombinant ns1 protein - by Bioz Stars, 2026-10
94/100 stars
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tick-borne encephalitis virus (tbev) (strain neudoerfl tbev-ns1, western subtype) tbev-ns1 partial gene orf cdna clone expression plasmid, n-myc tag  (Sino Biological)
N/A
Partial length Clone DNA of Tick-borne encephalitis virus (TBEV) (strain Neudoerfl TBEV-NS1, Western subtype) TBEV-NS1 with N terminal Myc tag.
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tbev ns1  (GenScript corporation)
90
GenScript corporation tbev ns1
In vitro characterization of <t>MVA-NS1.</t> (A) Homologous and intragenomic homologous (marker gene deletion) recombinations lead to production of MVA-NS1 with <t>TBEV</t> NS1 expression under transcriptional control of VACV late promotor psynII. (B) PCR products specific for the six major deletion sites inside the MVA genome performed on MVA-NS1 (1% agarose TBE gel) (I: 291 bp, II: 354 bp, III: 447 bp, IV: 502 bp, V: 603 bp, VI: 702 bp). Integration of the NS1 gene in deletion site III was confirmed (III: 1596 bp). (C) Immunostaining of wtMVA or MVA-NS1 infected HeLa cells (MOI 1). 24 hpi, cells were fixed with 4% PFA, for intracellular staining permeabilized with TritonX®-100 and immunostained against VACV D8 and TBEV NS1 (20x magnification). (D) Western blot analysis of whole cell lysate from HeLa cells infected with wtMVA or MVA-NS1 for 24 h (MOI 5). Blots were stained against TBEV NS1. For control, antibodies against VACV D8 and GAPDH were included. (E) Growth curves of wtMVA (black) and MVA-NS1 (gray) on primary CEF cells (dotted line) and HeLa cells (solid line) (MOI 0.05). Mann-Whitney test was used for statistical comparison between CEF and HeLa cells (* p<0.05).
Tbev Ns1, supplied by GenScript corporation, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/tbev+ns1/tbev+ns1/pmc10360051-67-15-26
Average 90 stars, based on 1 article reviews
tbev ns1 - by Bioz Stars, 2026-10
90/100 stars
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tick-borne encephalitis virus (tbev) (strain neudoerfl tbev-ns1, western subtype) tbev-ns1 partial gene orf cdna clone expression plasmid, c-ha tag  (Sino Biological)
N/A
Partial length Clone DNA of Tick-borne encephalitis virus (TBEV) (strain Neudoerfl TBEV-NS1, Western subtype) TBEV-NS1 with C terminal HA tag.
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tick-borne encephalitis virus (tbev) (strain neudoerfl tbev-ns1, western subtype) tbev-ns1 partial gene lentiviral orf cdna expression plasmid, c-gfps  (Sino Biological)
N/A
Partial length Clone DNA of Tick-borne encephalitis virus (TBEV) (strain Neudoerfl TBEV-NS1, Western subtype) TBEV-NS1.
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tick-borne encephalitis virus (tbev) (strain neudoerfl tbev-ns1, western subtype) tbev-ns1 partial gene orf cdna clone expression plasmid, c-ofpspark  (Sino Biological)
N/A
Partial length Clone DNA of Tick-borne encephalitis virus (TBEV) (strain Neudoerfl TBEV-NS1, Western subtype) TBEV-NS1 with C terminal OFPSpark / RFP tag.
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tick-borne encephalitis virus (tbev) (strain neudoerfl tbev-ns1, western subtype) tbev-ns1 partial gene orf cdna clone expression plasmid, c-flag tag  (Sino Biological)
N/A
Partial length Clone DNA of Tick-borne encephalitis virus (TBEV) (strain Neudoerfl TBEV-NS1, Western subtype) TBEV-NS1 with C terminal Flag tag.
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tick-borne encephalitis virus (tbev) (strain neudoerfl tbev-ns1, western subtype) tbev-ns1 partial gene orf cdna clone expression plasmid, c-his tag  (Sino Biological)
N/A
Partial length Clone DNA of Tick-borne encephalitis virus (TBEV) (strain Neudoerfl TBEV-NS1, Western subtype) TBEV-NS1 with C terminal His tag.
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tick-borne encephalitis virus (tbev) (strain neudoerfl tbev-ns1, western subtype) tbev-ns1 partial gene orf cdna clone expression plasmid, c-gfpspark  (Sino Biological)
N/A
Partial length Clone DNA of Tick-borne encephalitis virus (TBEV) (strain Neudoerfl TBEV-NS1, Western subtype) TBEV-NS1 with C terminal GFPSpark tag.
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tick-borne encephalitis virus (tbev) (strain neudoerfl tbev-ns1, western subtype) tbev-ns1 partial gene orf cdna clone in cloning vector  (Sino Biological)
N/A
Partial length Clone DNA of Tick-borne encephalitis virus (TBEV) (strain Neudoerfl TBEV-NS1, Western subtype) TBEV-NS1.
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tick-borne encephalitis virus (tbev) (strain neudoerfl tbev-ns1, western subtype) tbev-ns1 partial gene orf cdna clone expression plasmid, n-ha tag  (Sino Biological)
N/A
Partial length Clone DNA of Tick-borne encephalitis virus (TBEV) (strain Neudoerfl TBEV-NS1, Western subtype) TBEV-NS1 with N terminal HA tag.
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tick-borne encephalitis virus (tbev) (strain neudoerfl tbev-ns1, western subtype) tbev-ns1 partial natural gene orf cdna clone expression plasmid  (Sino Biological)
N/A
Partial length Clone DNA of Tick-borne encephalitis virus (TBEV) (strain Neudoerfl TBEV-NS1, Western subtype) TBEV-NS1.
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Image Search Results


In vitro characterization of MVA-NS1. (A) Homologous and intragenomic homologous (marker gene deletion) recombinations lead to production of MVA-NS1 with TBEV NS1 expression under transcriptional control of VACV late promotor psynII. (B) PCR products specific for the six major deletion sites inside the MVA genome performed on MVA-NS1 (1% agarose TBE gel) (I: 291 bp, II: 354 bp, III: 447 bp, IV: 502 bp, V: 603 bp, VI: 702 bp). Integration of the NS1 gene in deletion site III was confirmed (III: 1596 bp). (C) Immunostaining of wtMVA or MVA-NS1 infected HeLa cells (MOI 1). 24 hpi, cells were fixed with 4% PFA, for intracellular staining permeabilized with TritonX®-100 and immunostained against VACV D8 and TBEV NS1 (20x magnification). (D) Western blot analysis of whole cell lysate from HeLa cells infected with wtMVA or MVA-NS1 for 24 h (MOI 5). Blots were stained against TBEV NS1. For control, antibodies against VACV D8 and GAPDH were included. (E) Growth curves of wtMVA (black) and MVA-NS1 (gray) on primary CEF cells (dotted line) and HeLa cells (solid line) (MOI 0.05). Mann-Whitney test was used for statistical comparison between CEF and HeLa cells (* p<0.05).

Journal: Frontiers in Immunology

Article Title: Induction of humoral and cell-mediated immunity to the NS1 protein of TBEV with recombinant Influenza virus and MVA affords partial protection against lethal TBEV infection in mice

doi: 10.3389/fimmu.2023.1177324

Figure Lengend Snippet: In vitro characterization of MVA-NS1. (A) Homologous and intragenomic homologous (marker gene deletion) recombinations lead to production of MVA-NS1 with TBEV NS1 expression under transcriptional control of VACV late promotor psynII. (B) PCR products specific for the six major deletion sites inside the MVA genome performed on MVA-NS1 (1% agarose TBE gel) (I: 291 bp, II: 354 bp, III: 447 bp, IV: 502 bp, V: 603 bp, VI: 702 bp). Integration of the NS1 gene in deletion site III was confirmed (III: 1596 bp). (C) Immunostaining of wtMVA or MVA-NS1 infected HeLa cells (MOI 1). 24 hpi, cells were fixed with 4% PFA, for intracellular staining permeabilized with TritonX®-100 and immunostained against VACV D8 and TBEV NS1 (20x magnification). (D) Western blot analysis of whole cell lysate from HeLa cells infected with wtMVA or MVA-NS1 for 24 h (MOI 5). Blots were stained against TBEV NS1. For control, antibodies against VACV D8 and GAPDH were included. (E) Growth curves of wtMVA (black) and MVA-NS1 (gray) on primary CEF cells (dotted line) and HeLa cells (solid line) (MOI 0.05). Mann-Whitney test was used for statistical comparison between CEF and HeLa cells (* p<0.05).

Article Snippet: Plasmid encoding for the Kozak sequence followed by the TBEV E gene signal peptide and entire TBEV NS1 were synthesized (based on TBEV Neudoerfl, UniProtKB: P14336; GenScript Biotech Corp) and cloned into MVA transfer plasmid pIIIsynIIred under transcriptional control of VACV late promotor psynII ( ). pIIIsynIIred contains mCherry as marker gene which is flanked by short repetitive regions.

Techniques: In Vitro, Marker, Expressing, Control, Immunostaining, Infection, Staining, Western Blot, MANN-WHITNEY, Comparison

In vitro characterization of IAV-NS1. (A) Schematic representation of the NA-TBEV NS1 gene segment, NCR = non-coding region, ATG = start codon, *Stop = stop codon. (B) PCR products specific for IAV NA performed on I = rPR8, II = IAV-RBD, III = IAV-NS1 and IV = negative control (1% agarose TBE gel, M = 1 kb ladder). (C) Immunostaining of rPR8, IAV-RBD or IAV-NS1 infected MDCK cells (MOI 0.01). 24 hpi, cells were fixed with 4% PFA, for intracellular staining permeabilized with TritonX®-100 and immunostained against IAV NP or TBEV NS1 (10x magnification). (D) Western blot analysis of whole cell lysate from MDCK cells infected with rPR8, IAV-RBD or IAV-NS1 for 24 h (MOI 0.01). Blots were stained against TBEV NS1. For control, antibodies against IAV nucleoprotein (NP) and GAPDH were included. (E) Growth curves for rPR8 (black), IAV-RBD (gray) and IAV-NS1 (red) in the presence (solid line) or absence (dotted line) of eNA on MDCK cells (MOI 0.001). Mann-Whitney test was used for statistical comparison between rPR8 –eNA and IAV-RBD –eNA or IAV-NS1 –eNA, respectively (* p<0.05).

Journal: Frontiers in Immunology

Article Title: Induction of humoral and cell-mediated immunity to the NS1 protein of TBEV with recombinant Influenza virus and MVA affords partial protection against lethal TBEV infection in mice

doi: 10.3389/fimmu.2023.1177324

Figure Lengend Snippet: In vitro characterization of IAV-NS1. (A) Schematic representation of the NA-TBEV NS1 gene segment, NCR = non-coding region, ATG = start codon, *Stop = stop codon. (B) PCR products specific for IAV NA performed on I = rPR8, II = IAV-RBD, III = IAV-NS1 and IV = negative control (1% agarose TBE gel, M = 1 kb ladder). (C) Immunostaining of rPR8, IAV-RBD or IAV-NS1 infected MDCK cells (MOI 0.01). 24 hpi, cells were fixed with 4% PFA, for intracellular staining permeabilized with TritonX®-100 and immunostained against IAV NP or TBEV NS1 (10x magnification). (D) Western blot analysis of whole cell lysate from MDCK cells infected with rPR8, IAV-RBD or IAV-NS1 for 24 h (MOI 0.01). Blots were stained against TBEV NS1. For control, antibodies against IAV nucleoprotein (NP) and GAPDH were included. (E) Growth curves for rPR8 (black), IAV-RBD (gray) and IAV-NS1 (red) in the presence (solid line) or absence (dotted line) of eNA on MDCK cells (MOI 0.001). Mann-Whitney test was used for statistical comparison between rPR8 –eNA and IAV-RBD –eNA or IAV-NS1 –eNA, respectively (* p<0.05).

Article Snippet: Plasmid encoding for the Kozak sequence followed by the TBEV E gene signal peptide and entire TBEV NS1 were synthesized (based on TBEV Neudoerfl, UniProtKB: P14336; GenScript Biotech Corp) and cloned into MVA transfer plasmid pIIIsynIIred under transcriptional control of VACV late promotor psynII ( ). pIIIsynIIred contains mCherry as marker gene which is flanked by short repetitive regions.

Techniques: In Vitro, Negative Control, Immunostaining, Infection, Staining, Western Blot, Control, MANN-WHITNEY, Comparison

Antibody response against TBEV NS1. (A) Quantitative measurement of TBEV NS1-specific IgG by NS1 ELISA of sera samples collected 56 days post prime immunization. Results are reported as arbitrary units per ml (U/ml). Cut-off values were calculated according to the manufacturer’s instructions (1 U/ml). One-way ANOVA with Tukey’s multiple-comparison test was used for statistical analysis. Significances are shown for all NS1-specific vaccine groups (* p<0.05, **** p<0.0001). (B) Semi-quantitative measurement of TBEV NS1-specific antibodies by LIPS assay with mouse sera from day 0, 28 and 56 post prime immunization in relative light units (RLU). Mean values with SD are shown from 2-5 independent experiments (n=4 mice/group). Two-way ANOVA with Tukey’s multiple-comparison test was used for statistical analysis. Significances are shown for RLU values on d28 vs. d56 (solid line) and for all NS1-specific vaccine groups compared at d56 (dotted line) (** p<0.01, **** p<0.0001).

Journal: Frontiers in Immunology

Article Title: Induction of humoral and cell-mediated immunity to the NS1 protein of TBEV with recombinant Influenza virus and MVA affords partial protection against lethal TBEV infection in mice

doi: 10.3389/fimmu.2023.1177324

Figure Lengend Snippet: Antibody response against TBEV NS1. (A) Quantitative measurement of TBEV NS1-specific IgG by NS1 ELISA of sera samples collected 56 days post prime immunization. Results are reported as arbitrary units per ml (U/ml). Cut-off values were calculated according to the manufacturer’s instructions (1 U/ml). One-way ANOVA with Tukey’s multiple-comparison test was used for statistical analysis. Significances are shown for all NS1-specific vaccine groups (* p<0.05, **** p<0.0001). (B) Semi-quantitative measurement of TBEV NS1-specific antibodies by LIPS assay with mouse sera from day 0, 28 and 56 post prime immunization in relative light units (RLU). Mean values with SD are shown from 2-5 independent experiments (n=4 mice/group). Two-way ANOVA with Tukey’s multiple-comparison test was used for statistical analysis. Significances are shown for RLU values on d28 vs. d56 (solid line) and for all NS1-specific vaccine groups compared at d56 (dotted line) (** p<0.01, **** p<0.0001).

Article Snippet: Plasmid encoding for the Kozak sequence followed by the TBEV E gene signal peptide and entire TBEV NS1 were synthesized (based on TBEV Neudoerfl, UniProtKB: P14336; GenScript Biotech Corp) and cloned into MVA transfer plasmid pIIIsynIIred under transcriptional control of VACV late promotor psynII ( ). pIIIsynIIred contains mCherry as marker gene which is flanked by short repetitive regions.

Techniques: Enzyme-linked Immunosorbent Assay, Comparison, Lips Assay

T cell response against TBEV NS1. (A) For IFN-γ ELISpot, mouse splenocytes were restimulated overnight with TBEV NS1 peptide pools NS1 1-183 and NS1 173-352 . Mean values with SD are shown as IFN-γ spot-forming cells (SFC) per one million splenocytes after background subtraction. For statistical analysis, unpaired t-test was used (**p<0.01). (B) Calculated IFN-γ SFC per million splenocytes for total NS1 (NS1 1-183 + NS1 173-352 ). One-way ANOVA with Tukey’s multiple-comparison test was used for statistical analysis. Significances are only shown for NS1-vaccine groups (ns = not significant, ** p<0.01, **** p<0.0001). (C–G) Flow cytometric analysis of mouse splenocytes. Frequency of CD3 + subpopulations gated on CD4 + (C, D) and CD8 + (E–G) T cells positive for CD69, IFN-γ and Granzyme B (GrzB) upon restimulation with NS1 1-183 (black circle) or NS1 173-352 (unfilled square) (n = 4). Bars represent mean with SD. Data is shown after background subtraction.

Journal: Frontiers in Immunology

Article Title: Induction of humoral and cell-mediated immunity to the NS1 protein of TBEV with recombinant Influenza virus and MVA affords partial protection against lethal TBEV infection in mice

doi: 10.3389/fimmu.2023.1177324

Figure Lengend Snippet: T cell response against TBEV NS1. (A) For IFN-γ ELISpot, mouse splenocytes were restimulated overnight with TBEV NS1 peptide pools NS1 1-183 and NS1 173-352 . Mean values with SD are shown as IFN-γ spot-forming cells (SFC) per one million splenocytes after background subtraction. For statistical analysis, unpaired t-test was used (**p<0.01). (B) Calculated IFN-γ SFC per million splenocytes for total NS1 (NS1 1-183 + NS1 173-352 ). One-way ANOVA with Tukey’s multiple-comparison test was used for statistical analysis. Significances are only shown for NS1-vaccine groups (ns = not significant, ** p<0.01, **** p<0.0001). (C–G) Flow cytometric analysis of mouse splenocytes. Frequency of CD3 + subpopulations gated on CD4 + (C, D) and CD8 + (E–G) T cells positive for CD69, IFN-γ and Granzyme B (GrzB) upon restimulation with NS1 1-183 (black circle) or NS1 173-352 (unfilled square) (n = 4). Bars represent mean with SD. Data is shown after background subtraction.

Article Snippet: Plasmid encoding for the Kozak sequence followed by the TBEV E gene signal peptide and entire TBEV NS1 were synthesized (based on TBEV Neudoerfl, UniProtKB: P14336; GenScript Biotech Corp) and cloned into MVA transfer plasmid pIIIsynIIred under transcriptional control of VACV late promotor psynII ( ). pIIIsynIIred contains mCherry as marker gene which is flanked by short repetitive regions.

Techniques: Enzyme-linked Immunospot, Comparison

Protective efficacy of TBEV NS1-based vector constructs. Body weights and survival of BL6 mice immunized with controls (A, B) , IAV-NS1 or MVA-NS1 (C, D) , MVA-NS1/IAV-NS1 (E, F) or IAV-NS1/MVA-NS1 (G, F) after challenge infection with TBEV Neudoerfl. In (A) and (C) mean body weights are shown (n = 6). For (E) and (G) body weights from individual mice are shown. Weights of PBS mice are shown as mean (n = 6). (B, D, F) Kaplan-Meier curves were analyzed by log-rank test (*p<0.05, **p<0.01). (H) Pearson correlation of TBEV NS1-specific antibodies on day 56 before challenge infection measured by LIPS with days of survival post TBEV infection for all groups (FSME-IMMUN® group was excluded). Linear regression is depicted by gray line (r = 0.7964, p<0.0001).

Journal: Frontiers in Immunology

Article Title: Induction of humoral and cell-mediated immunity to the NS1 protein of TBEV with recombinant Influenza virus and MVA affords partial protection against lethal TBEV infection in mice

doi: 10.3389/fimmu.2023.1177324

Figure Lengend Snippet: Protective efficacy of TBEV NS1-based vector constructs. Body weights and survival of BL6 mice immunized with controls (A, B) , IAV-NS1 or MVA-NS1 (C, D) , MVA-NS1/IAV-NS1 (E, F) or IAV-NS1/MVA-NS1 (G, F) after challenge infection with TBEV Neudoerfl. In (A) and (C) mean body weights are shown (n = 6). For (E) and (G) body weights from individual mice are shown. Weights of PBS mice are shown as mean (n = 6). (B, D, F) Kaplan-Meier curves were analyzed by log-rank test (*p<0.05, **p<0.01). (H) Pearson correlation of TBEV NS1-specific antibodies on day 56 before challenge infection measured by LIPS with days of survival post TBEV infection for all groups (FSME-IMMUN® group was excluded). Linear regression is depicted by gray line (r = 0.7964, p<0.0001).

Article Snippet: Plasmid encoding for the Kozak sequence followed by the TBEV E gene signal peptide and entire TBEV NS1 were synthesized (based on TBEV Neudoerfl, UniProtKB: P14336; GenScript Biotech Corp) and cloned into MVA transfer plasmid pIIIsynIIred under transcriptional control of VACV late promotor psynII ( ). pIIIsynIIred contains mCherry as marker gene which is flanked by short repetitive regions.

Techniques: Plasmid Preparation, Construct, Infection

Viral loads in the periphery, CNS and GIT. Presence of TBEV RNA in spleen (A) , serum (B) , spinal cord (C) , brain (D) , ileum (E) and colon (F) was determined by performing real time RT-qPCR on cleared organ homogenates or serum from TBEV challenged mice sacrificed at 8 dpi (n = 6). Bars depict geometric means.

Journal: Frontiers in Immunology

Article Title: Induction of humoral and cell-mediated immunity to the NS1 protein of TBEV with recombinant Influenza virus and MVA affords partial protection against lethal TBEV infection in mice

doi: 10.3389/fimmu.2023.1177324

Figure Lengend Snippet: Viral loads in the periphery, CNS and GIT. Presence of TBEV RNA in spleen (A) , serum (B) , spinal cord (C) , brain (D) , ileum (E) and colon (F) was determined by performing real time RT-qPCR on cleared organ homogenates or serum from TBEV challenged mice sacrificed at 8 dpi (n = 6). Bars depict geometric means.

Article Snippet: Plasmid encoding for the Kozak sequence followed by the TBEV E gene signal peptide and entire TBEV NS1 were synthesized (based on TBEV Neudoerfl, UniProtKB: P14336; GenScript Biotech Corp) and cloned into MVA transfer plasmid pIIIsynIIred under transcriptional control of VACV late promotor psynII ( ). pIIIsynIIred contains mCherry as marker gene which is flanked by short repetitive regions.

Techniques: Quantitative RT-PCR

Histological and immunohistochemical changes in the olfactory bulb at 8 dpi. (A–H) Hematoxylin and eosin (H&E) stained sections of the olfactory bulb of a mouse treated with PBS (A) or vaccinated with FSME-IMMUN® (B) , wtMVA (C) , IAV-RBD (D) , IAV-NS1 (E) , MVA-NS1 (F) , MVA-NS1/IAV-NS1 (G) or IAV-NS1/MVA-NS1 (H) , respectively. (A) Olfactory bulb of the PBS-treated mouse displays marked cellular necrosis with karyorrhectic, karyolytic and pyknotic cells (insert, arrowhead) and shrunken, hypereosinophilic, triangular shaped necrotic neurons as well as inflammatory cell infiltrates. (B–H) In FSME-IMMUN®- (B) , wtMVA- (C) , IAV-NS1- (E) , MVA-NS1- (F) , MVA-NS1/IAV-NS1- (G) or IAV-NS1/MVA-NS1- (H) vaccinated mice, no significant microscopic lesions within the olfactory bulb are visible. In the IAV-RBD (D) vaccinated mouse, single necrotic cells are present. (I–P) Immunohistochemistry for TBEV E antigen of the olfactory bulb of a mouse treated with PBS (I) or vaccinated with FSME-IMMUN® (J) , wtMVA (K) , IAV-RBD (L) , IAV-NS1 (M) , MVA-NS1 (N) , MVA-NS1/IAV-NS1 (O) or IAV-NS1/MVA-NS1 (P) , respectively. (I) Immunohistochemically, a cytoplasmic TBEV immunoreactivity (arrowhead) is present in multiple cells of the olfactory bulb from the PBS-treated mouse. (J–P) Olfactory bulbs of FSME-IMMUN®- (J) , wtMVA- (K) , IAV-NS1- (M) , MVA-NS1- (N) , MVA-NS1/IAV-NS1- (O) or IAV-NS1/MVA-NS1- (P) vaccinated mice do not show immunoreactivity for TBEV, while the IAV-RBD- (N) vaccinated mouse shows single TBEV-positive cells (arrowhead). Scale bars: 20µm.

Journal: Frontiers in Immunology

Article Title: Induction of humoral and cell-mediated immunity to the NS1 protein of TBEV with recombinant Influenza virus and MVA affords partial protection against lethal TBEV infection in mice

doi: 10.3389/fimmu.2023.1177324

Figure Lengend Snippet: Histological and immunohistochemical changes in the olfactory bulb at 8 dpi. (A–H) Hematoxylin and eosin (H&E) stained sections of the olfactory bulb of a mouse treated with PBS (A) or vaccinated with FSME-IMMUN® (B) , wtMVA (C) , IAV-RBD (D) , IAV-NS1 (E) , MVA-NS1 (F) , MVA-NS1/IAV-NS1 (G) or IAV-NS1/MVA-NS1 (H) , respectively. (A) Olfactory bulb of the PBS-treated mouse displays marked cellular necrosis with karyorrhectic, karyolytic and pyknotic cells (insert, arrowhead) and shrunken, hypereosinophilic, triangular shaped necrotic neurons as well as inflammatory cell infiltrates. (B–H) In FSME-IMMUN®- (B) , wtMVA- (C) , IAV-NS1- (E) , MVA-NS1- (F) , MVA-NS1/IAV-NS1- (G) or IAV-NS1/MVA-NS1- (H) vaccinated mice, no significant microscopic lesions within the olfactory bulb are visible. In the IAV-RBD (D) vaccinated mouse, single necrotic cells are present. (I–P) Immunohistochemistry for TBEV E antigen of the olfactory bulb of a mouse treated with PBS (I) or vaccinated with FSME-IMMUN® (J) , wtMVA (K) , IAV-RBD (L) , IAV-NS1 (M) , MVA-NS1 (N) , MVA-NS1/IAV-NS1 (O) or IAV-NS1/MVA-NS1 (P) , respectively. (I) Immunohistochemically, a cytoplasmic TBEV immunoreactivity (arrowhead) is present in multiple cells of the olfactory bulb from the PBS-treated mouse. (J–P) Olfactory bulbs of FSME-IMMUN®- (J) , wtMVA- (K) , IAV-NS1- (M) , MVA-NS1- (N) , MVA-NS1/IAV-NS1- (O) or IAV-NS1/MVA-NS1- (P) vaccinated mice do not show immunoreactivity for TBEV, while the IAV-RBD- (N) vaccinated mouse shows single TBEV-positive cells (arrowhead). Scale bars: 20µm.

Article Snippet: Plasmid encoding for the Kozak sequence followed by the TBEV E gene signal peptide and entire TBEV NS1 were synthesized (based on TBEV Neudoerfl, UniProtKB: P14336; GenScript Biotech Corp) and cloned into MVA transfer plasmid pIIIsynIIred under transcriptional control of VACV late promotor psynII ( ). pIIIsynIIred contains mCherry as marker gene which is flanked by short repetitive regions.

Techniques: Immunohistochemical staining, Staining, Immunohistochemistry

Histological and immunohistochemical analysis of colon at 8 dpi. (A–H) Hematoxylin and eosin (H&E) stained sections of the colon of a mouse treated with PBS (A) or vaccinated with FSME-IMMUN®, (B) , wtMVA (C) , IAV-RBD (D) , IAV-NS1 (E) , MVA-NS1 (F) , MVA-NS1/IAV-NS1 (G) or IAV-NS1/MVA-NS1 (H) , respectively. (A) Colon of the PBS-treated mouse displays minor to mild hypercellularity and cellular necrosis in the myenteric (arrowhead) and submucosal plexus (asterisk). (B–F) Colon of the FSME-IMMUN®- (B) , wtMVA- (C) , IAV-RBD- (D) , and MVA-NS1- (F) vaccinated mice show minimal to moderate histopathological changes mainly characterized by hypercellularity/inflammatory cell infiltrates in the myenteric and submucosal plexus. (E–H) In the IAV-NS1- (E) , MVA-NS1/IAV-NS1- (G) and IAV-NS1/MVA-NS1- (H) vaccinated mice, no histopathological changes in the plexi of the colon are observed. (I–P) Immunohistochemistry for TBEV E antigen of the colon of a mouse treated with PBS (I) or vaccinated with FSME-IMMUN® (J) , wtMVA (K) , IAV-RBD (L) , IAV-NS1 (M) , MVA-NS1 (N) , MVA-NS1/IAV-NS1 (O) or IAV-NS1/MVA-NS1 (P) , respectively. (I, K, L) Immunohistochemically, a cytoplasmic TBEV immunoreactivity is present in cells of the myenteric plexus (arrowhead) of the colon from the PBS-treated mouse (I) , the wtMVA-vaccinated mouse (K) and the IAV-RBD-vaccinated mouse (L) . A single TBEV-antigen positive cell is visible in the submucosal plexus of the colon from the MVA-NS1/IAV-NS1-vaccinated mouse ( O , asterisk). No TBEV-antigen positive cells are detectable in the submucosal (asterisk) or myenteric plexus (arrowhead) of FSME-IMMUN®- (J) , IAV-NS1- (M) , MVA-NS1- (N) and IAV-NS1/MVA-NS1-vaccinated (P) mice. Scale bars (A, B, D–H) : 20µm; Scale bar: (C, I–P) : 50µm.

Journal: Frontiers in Immunology

Article Title: Induction of humoral and cell-mediated immunity to the NS1 protein of TBEV with recombinant Influenza virus and MVA affords partial protection against lethal TBEV infection in mice

doi: 10.3389/fimmu.2023.1177324

Figure Lengend Snippet: Histological and immunohistochemical analysis of colon at 8 dpi. (A–H) Hematoxylin and eosin (H&E) stained sections of the colon of a mouse treated with PBS (A) or vaccinated with FSME-IMMUN®, (B) , wtMVA (C) , IAV-RBD (D) , IAV-NS1 (E) , MVA-NS1 (F) , MVA-NS1/IAV-NS1 (G) or IAV-NS1/MVA-NS1 (H) , respectively. (A) Colon of the PBS-treated mouse displays minor to mild hypercellularity and cellular necrosis in the myenteric (arrowhead) and submucosal plexus (asterisk). (B–F) Colon of the FSME-IMMUN®- (B) , wtMVA- (C) , IAV-RBD- (D) , and MVA-NS1- (F) vaccinated mice show minimal to moderate histopathological changes mainly characterized by hypercellularity/inflammatory cell infiltrates in the myenteric and submucosal plexus. (E–H) In the IAV-NS1- (E) , MVA-NS1/IAV-NS1- (G) and IAV-NS1/MVA-NS1- (H) vaccinated mice, no histopathological changes in the plexi of the colon are observed. (I–P) Immunohistochemistry for TBEV E antigen of the colon of a mouse treated with PBS (I) or vaccinated with FSME-IMMUN® (J) , wtMVA (K) , IAV-RBD (L) , IAV-NS1 (M) , MVA-NS1 (N) , MVA-NS1/IAV-NS1 (O) or IAV-NS1/MVA-NS1 (P) , respectively. (I, K, L) Immunohistochemically, a cytoplasmic TBEV immunoreactivity is present in cells of the myenteric plexus (arrowhead) of the colon from the PBS-treated mouse (I) , the wtMVA-vaccinated mouse (K) and the IAV-RBD-vaccinated mouse (L) . A single TBEV-antigen positive cell is visible in the submucosal plexus of the colon from the MVA-NS1/IAV-NS1-vaccinated mouse ( O , asterisk). No TBEV-antigen positive cells are detectable in the submucosal (asterisk) or myenteric plexus (arrowhead) of FSME-IMMUN®- (J) , IAV-NS1- (M) , MVA-NS1- (N) and IAV-NS1/MVA-NS1-vaccinated (P) mice. Scale bars (A, B, D–H) : 20µm; Scale bar: (C, I–P) : 50µm.

Article Snippet: Plasmid encoding for the Kozak sequence followed by the TBEV E gene signal peptide and entire TBEV NS1 were synthesized (based on TBEV Neudoerfl, UniProtKB: P14336; GenScript Biotech Corp) and cloned into MVA transfer plasmid pIIIsynIIred under transcriptional control of VACV late promotor psynII ( ). pIIIsynIIred contains mCherry as marker gene which is flanked by short repetitive regions.

Techniques: Immunohistochemical staining, Staining, Immunohistochemistry