zika virus envelope protein Search Results


zikv envelop protein  (ATCC)
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ATCC zikv envelop protein
Zikv Envelop Protein, supplied by ATCC, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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zika virus envelope protein  (Native Antigen Inc)
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Native Antigen Inc zika virus envelope protein
Zika Virus Envelope 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
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zika virus  (Sino Biological)
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Sino Biological zika virus
Advances and potential application of biosensors in pharmaceutical sciences
Zika Virus, supplied by Sino Biological, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti zediii antibody  (Sino Biological)
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Sino Biological anti zediii antibody
Advances and potential application of biosensors in pharmaceutical sciences
Anti Zediii Antibody, supplied by Sino Biological, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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recombinant zikv envelope protein  (Sino Biological)
94
Sino Biological recombinant zikv envelope protein
<t>ZIKV</t> infection of olfactory tissues in suckling ICR mice. ( A to D ) Seven-day-old ICR suckling mice were infected intracerebrally with 10 PFU of Zika virus (ZIKV) ( A ). Blood samples were collected at 28 days after ZIKV infection or mock treatment. ZIKV-specific IgG antibody was determined by ELISA plates coated with soluble <t>ZIKV</t> <t>Envelope</t> protein. (n = 21) ( B ), and serum levels of viral RNA were detected by qRT–PCR analysis (n = 5) ( C ). The olfactory function of infected and mock-treated mice was assessed by a buried food pellet test 4–6 weeks post-infection and 4 weeks post inoculation, respectively (n = 18) ( D ). Data are presented as median and interquartile range and were analyzed using Mann Whitney U test (B) and Kruskal–Wallis with Dunn's multiple comparisons test (C, D). ( E and F ) Three-day-old ICR suckling mice were infected intracerebrally with 10 3 PFU of ZIKV, mice were sacrificed at different time points, the OM, OB and brain tissues (n = 3) were collected to analyze ZIKV RNA levels by qRT–PCR analysis ( E ) and viral loads by in situ RNAscope hybridization, Scale bar, 50 μM ( F ). The limit of detection (LOD) is indicated by the dotted line. The data are presented as the mean ± SD and were analyzed with Ordinary one-way ANOVA test. p values are indicated by ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001.
Recombinant Zikv Envelope Protein, supplied by Sino Biological, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti zikv e protein rabbit  (Sino Biological)
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Sino Biological anti zikv e protein rabbit
Antibodies used for immunofluorescence assay (IFA).
Anti Zikv E Protein Rabbit, supplied by Sino Biological, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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invivomab anti mouse ifnar 1 antibody  (Sino Biological)
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Sino Biological invivomab anti mouse ifnar 1 antibody
Antibodies used for immunofluorescence assay (IFA).
Invivomab Anti Mouse Ifnar 1 Antibody, supplied by Sino Biological, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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elisa test  (Sino Biological)
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Sino Biological elisa test
Antibodies used for immunofluorescence assay (IFA).
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domain iii cat 40543 v08y  (Sino Biological)
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Sino Biological domain iii cat 40543 v08y
Antibodies used for immunofluorescence assay (IFA).
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recombinant proteins zika virus full length envelope e protein aviva systems biology opma04848 zika virus edi edii peptides  (Aviva Systems)
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Aviva Systems recombinant proteins zika virus full length envelope e protein aviva systems biology opma04848 zika virus edi edii peptides
Figure 1. <t>ZIKV</t> TCI vaccine design and antigen expression (A) Schematic diagram of plasmid design. The gene sequence encoding for the NS3 protein was split into three parts (denoted 1, 2, and 3). The 30 nucleotide bases before and after any cleaved sequence were placed in front of each region to preserve any epitopes that may have been disrupted. The ORF gene en- coding for a mouse/human monomer of ubiquitin (Ub) was placed immediately upstream of the rearranged NS3 sequence. A glycine at the 76th residue was modified to encode an alanine to enhance the stability of the Ub/NS3 complex. (B) 293T cells were transfected with the rearranged Ub/NS3 plasmid overnight. The cells were allowed to stably express plasmid for 36 h. After this period, MG132 was added overnight. The cell lysate was analyzed via western blot for the expression of NS3 and beta-actin. A representative blot from three ex- periments is shown. (C) Total band density of western blot was quantified using ImageJ and analyzed using GraphPad (Prism). The data are represented as mean ± SEM (n = 2 for each experiment, three experiments total). *p < 0.05; **p < 0.01.
Recombinant Proteins Zika Virus Full Length Envelope E Protein Aviva Systems Biology Opma04848 Zika Virus Edi Edii Peptides, supplied by Aviva Systems, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti zikv e protein antibody  (Sino Biological)
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Sino Biological anti zikv e protein antibody
Figure 1. <t>ZIKV</t> TCI vaccine design and antigen expression (A) Schematic diagram of plasmid design. The gene sequence encoding for the NS3 protein was split into three parts (denoted 1, 2, and 3). The 30 nucleotide bases before and after any cleaved sequence were placed in front of each region to preserve any epitopes that may have been disrupted. The ORF gene en- coding for a mouse/human monomer of ubiquitin (Ub) was placed immediately upstream of the rearranged NS3 sequence. A glycine at the 76th residue was modified to encode an alanine to enhance the stability of the Ub/NS3 complex. (B) 293T cells were transfected with the rearranged Ub/NS3 plasmid overnight. The cells were allowed to stably express plasmid for 36 h. After this period, MG132 was added overnight. The cell lysate was analyzed via western blot for the expression of NS3 and beta-actin. A representative blot from three ex- periments is shown. (C) Total band density of western blot was quantified using ImageJ and analyzed using GraphPad (Prism). The data are represented as mean ± SEM (n = 2 for each experiment, three experiments total). *p < 0.05; **p < 0.01.
Anti Zikv E Protein Antibody, supplied by Sino Biological, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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pcmv zikaenv  (Sino Biological)
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Sino Biological pcmv zikaenv
Figure 1. <t>ZIKV</t> TCI vaccine design and antigen expression (A) Schematic diagram of plasmid design. The gene sequence encoding for the NS3 protein was split into three parts (denoted 1, 2, and 3). The 30 nucleotide bases before and after any cleaved sequence were placed in front of each region to preserve any epitopes that may have been disrupted. The ORF gene en- coding for a mouse/human monomer of ubiquitin (Ub) was placed immediately upstream of the rearranged NS3 sequence. A glycine at the 76th residue was modified to encode an alanine to enhance the stability of the Ub/NS3 complex. (B) 293T cells were transfected with the rearranged Ub/NS3 plasmid overnight. The cells were allowed to stably express plasmid for 36 h. After this period, MG132 was added overnight. The cell lysate was analyzed via western blot for the expression of NS3 and beta-actin. A representative blot from three ex- periments is shown. (C) Total band density of western blot was quantified using ImageJ and analyzed using GraphPad (Prism). The data are represented as mean ± SEM (n = 2 for each experiment, three experiments total). *p < 0.05; **p < 0.01.
Pcmv Zikaenv, supplied by Sino Biological, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


Advances and potential application of biosensors in pharmaceutical sciences

Journal: Applied Microbiology and Biotechnology

Article Title: Recent advances in the potential applications of luminescence-based, SPR-based, and carbon-based biosensors

doi: 10.1007/s00253-022-11901-6

Figure Lengend Snippet: Advances and potential application of biosensors in pharmaceutical sciences

Article Snippet: Electrochemical immunosensor , Zika virus (envelope protein) , 10 pM , Zika virus (strain Zika SPH2015) envelope protein (ZIKV-E) ELISA Pair Set; Sino Biological Inc. (LOD = 125 pg/mL) , Promising clinical application for early-stage diagnostics of the virus, operation time around 40 min , Kaushik et al. ( ) .

Techniques: Förster Resonance Energy Transfer, Imaging, SPR Assay, Electrochemiluminescence, MicroChIP Assay, In Vitro, Infection, Amplification, Isolation, Biomarker Assay, Sequencing

Summarization of LOD, advances of the abovementioned biosensors, and their comparison with common methods of detection

Journal: Applied Microbiology and Biotechnology

Article Title: Recent advances in the potential applications of luminescence-based, SPR-based, and carbon-based biosensors

doi: 10.1007/s00253-022-11901-6

Figure Lengend Snippet: Summarization of LOD, advances of the abovementioned biosensors, and their comparison with common methods of detection

Article Snippet: Electrochemical immunosensor , Zika virus (envelope protein) , 10 pM , Zika virus (strain Zika SPH2015) envelope protein (ZIKV-E) ELISA Pair Set; Sino Biological Inc. (LOD = 125 pg/mL) , Promising clinical application for early-stage diagnostics of the virus, operation time around 40 min , Kaushik et al. ( ) .

Techniques: Glutamate Assay, Enzyme-linked Immunosorbent Assay, Diagnostic Assay, Sequencing, Simultaneous Assay, Biomarker Assay, Fluorescence, In Situ Hybridization

ZIKV infection of olfactory tissues in suckling ICR mice. ( A to D ) Seven-day-old ICR suckling mice were infected intracerebrally with 10 PFU of Zika virus (ZIKV) ( A ). Blood samples were collected at 28 days after ZIKV infection or mock treatment. ZIKV-specific IgG antibody was determined by ELISA plates coated with soluble ZIKV Envelope protein. (n = 21) ( B ), and serum levels of viral RNA were detected by qRT–PCR analysis (n = 5) ( C ). The olfactory function of infected and mock-treated mice was assessed by a buried food pellet test 4–6 weeks post-infection and 4 weeks post inoculation, respectively (n = 18) ( D ). Data are presented as median and interquartile range and were analyzed using Mann Whitney U test (B) and Kruskal–Wallis with Dunn's multiple comparisons test (C, D). ( E and F ) Three-day-old ICR suckling mice were infected intracerebrally with 10 3 PFU of ZIKV, mice were sacrificed at different time points, the OM, OB and brain tissues (n = 3) were collected to analyze ZIKV RNA levels by qRT–PCR analysis ( E ) and viral loads by in situ RNAscope hybridization, Scale bar, 50 μM ( F ). The limit of detection (LOD) is indicated by the dotted line. The data are presented as the mean ± SD and were analyzed with Ordinary one-way ANOVA test. p values are indicated by ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001.

Journal: eBioMedicine

Article Title: Zika virus leads to olfactory disorders in mice by targeting olfactory ensheathing cells

doi: 10.1016/j.ebiom.2023.104457

Figure Lengend Snippet: ZIKV infection of olfactory tissues in suckling ICR mice. ( A to D ) Seven-day-old ICR suckling mice were infected intracerebrally with 10 PFU of Zika virus (ZIKV) ( A ). Blood samples were collected at 28 days after ZIKV infection or mock treatment. ZIKV-specific IgG antibody was determined by ELISA plates coated with soluble ZIKV Envelope protein. (n = 21) ( B ), and serum levels of viral RNA were detected by qRT–PCR analysis (n = 5) ( C ). The olfactory function of infected and mock-treated mice was assessed by a buried food pellet test 4–6 weeks post-infection and 4 weeks post inoculation, respectively (n = 18) ( D ). Data are presented as median and interquartile range and were analyzed using Mann Whitney U test (B) and Kruskal–Wallis with Dunn's multiple comparisons test (C, D). ( E and F ) Three-day-old ICR suckling mice were infected intracerebrally with 10 3 PFU of ZIKV, mice were sacrificed at different time points, the OM, OB and brain tissues (n = 3) were collected to analyze ZIKV RNA levels by qRT–PCR analysis ( E ) and viral loads by in situ RNAscope hybridization, Scale bar, 50 μM ( F ). The limit of detection (LOD) is indicated by the dotted line. The data are presented as the mean ± SD and were analyzed with Ordinary one-way ANOVA test. p values are indicated by ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001.

Article Snippet: Briefly, 96-well polysorb enzyme-linked immunosorbent assay plates (Nunc, USA) coated with 100 ng per well concentration of recombinant ZIKV envelope protein (Cat: 40543-V08B4, Sino Biological, Beijing, China) were used.

Techniques: Infection, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, MANN-WHITNEY, In Situ, Hybridization

Antibodies used for immunofluorescence assay (IFA).

Journal: Viruses

Article Title: A Productive Expression Platform Derived from Host-Restricted Eilat Virus: Its Extensive Validation and Novel Strategy

doi: 10.3390/v13040660

Figure Lengend Snippet: Antibodies used for immunofluorescence assay (IFA).

Article Snippet: Anti-ZIKV E protein (rabbit) , Figure 8 , Sino Biological , 40543-R029 , .

Techniques: Immunofluorescence

Figure 1. ZIKV TCI vaccine design and antigen expression (A) Schematic diagram of plasmid design. The gene sequence encoding for the NS3 protein was split into three parts (denoted 1, 2, and 3). The 30 nucleotide bases before and after any cleaved sequence were placed in front of each region to preserve any epitopes that may have been disrupted. The ORF gene en- coding for a mouse/human monomer of ubiquitin (Ub) was placed immediately upstream of the rearranged NS3 sequence. A glycine at the 76th residue was modified to encode an alanine to enhance the stability of the Ub/NS3 complex. (B) 293T cells were transfected with the rearranged Ub/NS3 plasmid overnight. The cells were allowed to stably express plasmid for 36 h. After this period, MG132 was added overnight. The cell lysate was analyzed via western blot for the expression of NS3 and beta-actin. A representative blot from three ex- periments is shown. (C) Total band density of western blot was quantified using ImageJ and analyzed using GraphPad (Prism). The data are represented as mean ± SEM (n = 2 for each experiment, three experiments total). *p < 0.05; **p < 0.01.

Journal: Cell reports

Article Title: A vaccine inducing solely cytotoxic T lymphocytes fully prevents Zika virus infection and fetal damage.

doi: 10.1016/j.celrep.2021.109107

Figure Lengend Snippet: Figure 1. ZIKV TCI vaccine design and antigen expression (A) Schematic diagram of plasmid design. The gene sequence encoding for the NS3 protein was split into three parts (denoted 1, 2, and 3). The 30 nucleotide bases before and after any cleaved sequence were placed in front of each region to preserve any epitopes that may have been disrupted. The ORF gene en- coding for a mouse/human monomer of ubiquitin (Ub) was placed immediately upstream of the rearranged NS3 sequence. A glycine at the 76th residue was modified to encode an alanine to enhance the stability of the Ub/NS3 complex. (B) 293T cells were transfected with the rearranged Ub/NS3 plasmid overnight. The cells were allowed to stably express plasmid for 36 h. After this period, MG132 was added overnight. The cell lysate was analyzed via western blot for the expression of NS3 and beta-actin. A representative blot from three ex- periments is shown. (C) Total band density of western blot was quantified using ImageJ and analyzed using GraphPad (Prism). The data are represented as mean ± SEM (n = 2 for each experiment, three experiments total). *p < 0.05; **p < 0.01.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-mouse CD4 (IgG2b) antibody Bio X Cell BP0003-1; RRID:AB_1107636 Anti-mouse CD8a (IgG2b) antibody Bio X Cell BP0061; RRID:AB_1125541 Rat IgG2b isotype control antibody Bio X Cell BP0090; RRID:AB_1107780 Anti-mouse-CD8a (IgG2a) antibody Bio X Cell BE0004-1; RRID:AB_1107671 Rat IgG2a isotype control antibody Bio X Cell BE0089; RRID:AB_1107769 Anti-mouse IFNAR1 (MAR1-5A3) antibody Leinco I-401; RRID:AB_2491621 ZIKV EDIII-specific human antibody, ZV-64 Absolute Antibody Ab00811-10.0 Rabbit anti-vimentin antibody Abcam Ab92547; RRID:AB_10562134 Rabbit anti-active caspase-3 antibody Abcam Ab2302; RRID:AB_302962 FITC-anti-human antibody Sigma 9512-1mL; RRID:AB_259808 Anti-rabbit Alexa Fluor 647 antibody Abcam Ab150075; RRID:AB_2752244 FITC-anti-mouse antibody Biolegend 406001; RRID:AB_315029 Anti-flavivirus 4G2 antibody BEI Resources NR-50327 PerCP-Cy5.5-anti-mouse CD8 antibody BD Biosciences 551162; RRID:AB_394081 FITC-anti-mouse IL-2 antibody BD Biosciences 554427; RRID:AB_395385 FITC-anti-mouse CD4 antibody BD Biosciences 553729; RRID:AB_395013 PE-anti-mouse IFN-g antibody BD Biosciences 554412; RRID:AB_395376 Brilliant Violet 421-anti-mouse TNF-a antibody Biolegend 506328; RRID:AB_2562902 HRP-conjugated anti-mouse IgG-Fab antibody Sigma a9917-1mL; RRID:AB_258476 Anti-ZIKV NS3 antibody Genetex GTX133309; RRID:AB_2756864 Bacterial and virus strains Zika virus (human strain R103451 (2015/ Honduras)) BEI Resources NR-50355 Zika virus (human strain PAN2016 (2016/ Panama)) BEI Resources NR-50210 Chemicals, peptides, and recombinant proteins Zika virus full-length envelope (E) protein Aviva Systems Biology OPMA04848 Zika virus EDI/EDII peptides (see below peptide 1-5) BEI Resources NR-50553 Peptide 1: IGVSNRDFVEGMSGG BEI Resources N/A Peptide 2: TWVDVVLEHGGCVTV BEI Resources N/A Peptide 3: MAQDKPTVDIELVTT BEI Resources N/A Peptide 4: VDRGWGNGCGLFGKG BEI Resources N/A Peptide 5: VVLGSQEGAVHTALA BEI Resources N/A Zika virus NS3 overlapping peptides (see below peptide 6-15) GenScript N/A Peptide 6: AETDEDHAHWLEARM GenScript N/A Peptide 7: HAHWLEARMLLDNIY GenScript N/A Peptide 8: ARMLLDNIYLQDGLI GenScript N/A Peptide 9: NIYLQDGLIASLYRP GenScript N/A Peptide 10: GLIASLYRPEADKVA GenScript N/A Peptide 11: YRPEADKVAAIEGEF GenScript N/A (Continued on next page) e1 Cell Reports 35, 109107, May 11, 2021

Techniques: Expressing, Plasmid Preparation, Sequencing, Ubiquitin Proteomics, Residue, Transfection, Stable Transfection, Western Blot

Figure 2. ZIKV TCI-DNA vaccine protected adult male and female Ifnar1/ mice against ZIKV challenge with complete survival and reduced viral titers, including reproductive organs Equal numbers of male and female Ifnar1/ mice were immunized with ZIKV E ectodomain, EDI/II peptides, TCI-DNA, or PBS control, and sera were collected at 10 days post-second immunization. (A and B) At 13 days post-second immunization, the mice were challenged (i.p.) with ZIKV (strain R103451, 103 PFU/mouse) and recorded for weight (A) and survival (B) daily for 14 days (n = 6). (C) In a separate experiment, the immunized Ifnar1/ mice were challenged with ZIKV (strain PAN2016, 103 PFU/mouse), and 6 days later, sera and tissues (including brain, kidney, lung, heart, liver, spleen, muscle, and testis) were collected for detection of viral titers using plaque-forming assay. The detection limit was 25 PFU/ml (for sera), 12.5 PFU/g (for brain), 20 PFU/g (for kidney, spleen, and muscle), and 25 PFU/g (for lung, heart, liver, and testis). The data are represented as mean ± SEM (n = 3 for testis and n = 6 for other groups). *p < 0.05; **p < 0.01; ***p < 0.001.

Journal: Cell reports

Article Title: A vaccine inducing solely cytotoxic T lymphocytes fully prevents Zika virus infection and fetal damage.

doi: 10.1016/j.celrep.2021.109107

Figure Lengend Snippet: Figure 2. ZIKV TCI-DNA vaccine protected adult male and female Ifnar1/ mice against ZIKV challenge with complete survival and reduced viral titers, including reproductive organs Equal numbers of male and female Ifnar1/ mice were immunized with ZIKV E ectodomain, EDI/II peptides, TCI-DNA, or PBS control, and sera were collected at 10 days post-second immunization. (A and B) At 13 days post-second immunization, the mice were challenged (i.p.) with ZIKV (strain R103451, 103 PFU/mouse) and recorded for weight (A) and survival (B) daily for 14 days (n = 6). (C) In a separate experiment, the immunized Ifnar1/ mice were challenged with ZIKV (strain PAN2016, 103 PFU/mouse), and 6 days later, sera and tissues (including brain, kidney, lung, heart, liver, spleen, muscle, and testis) were collected for detection of viral titers using plaque-forming assay. The detection limit was 25 PFU/ml (for sera), 12.5 PFU/g (for brain), 20 PFU/g (for kidney, spleen, and muscle), and 25 PFU/g (for lung, heart, liver, and testis). The data are represented as mean ± SEM (n = 3 for testis and n = 6 for other groups). *p < 0.05; **p < 0.01; ***p < 0.001.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-mouse CD4 (IgG2b) antibody Bio X Cell BP0003-1; RRID:AB_1107636 Anti-mouse CD8a (IgG2b) antibody Bio X Cell BP0061; RRID:AB_1125541 Rat IgG2b isotype control antibody Bio X Cell BP0090; RRID:AB_1107780 Anti-mouse-CD8a (IgG2a) antibody Bio X Cell BE0004-1; RRID:AB_1107671 Rat IgG2a isotype control antibody Bio X Cell BE0089; RRID:AB_1107769 Anti-mouse IFNAR1 (MAR1-5A3) antibody Leinco I-401; RRID:AB_2491621 ZIKV EDIII-specific human antibody, ZV-64 Absolute Antibody Ab00811-10.0 Rabbit anti-vimentin antibody Abcam Ab92547; RRID:AB_10562134 Rabbit anti-active caspase-3 antibody Abcam Ab2302; RRID:AB_302962 FITC-anti-human antibody Sigma 9512-1mL; RRID:AB_259808 Anti-rabbit Alexa Fluor 647 antibody Abcam Ab150075; RRID:AB_2752244 FITC-anti-mouse antibody Biolegend 406001; RRID:AB_315029 Anti-flavivirus 4G2 antibody BEI Resources NR-50327 PerCP-Cy5.5-anti-mouse CD8 antibody BD Biosciences 551162; RRID:AB_394081 FITC-anti-mouse IL-2 antibody BD Biosciences 554427; RRID:AB_395385 FITC-anti-mouse CD4 antibody BD Biosciences 553729; RRID:AB_395013 PE-anti-mouse IFN-g antibody BD Biosciences 554412; RRID:AB_395376 Brilliant Violet 421-anti-mouse TNF-a antibody Biolegend 506328; RRID:AB_2562902 HRP-conjugated anti-mouse IgG-Fab antibody Sigma a9917-1mL; RRID:AB_258476 Anti-ZIKV NS3 antibody Genetex GTX133309; RRID:AB_2756864 Bacterial and virus strains Zika virus (human strain R103451 (2015/ Honduras)) BEI Resources NR-50355 Zika virus (human strain PAN2016 (2016/ Panama)) BEI Resources NR-50210 Chemicals, peptides, and recombinant proteins Zika virus full-length envelope (E) protein Aviva Systems Biology OPMA04848 Zika virus EDI/EDII peptides (see below peptide 1-5) BEI Resources NR-50553 Peptide 1: IGVSNRDFVEGMSGG BEI Resources N/A Peptide 2: TWVDVVLEHGGCVTV BEI Resources N/A Peptide 3: MAQDKPTVDIELVTT BEI Resources N/A Peptide 4: VDRGWGNGCGLFGKG BEI Resources N/A Peptide 5: VVLGSQEGAVHTALA BEI Resources N/A Zika virus NS3 overlapping peptides (see below peptide 6-15) GenScript N/A Peptide 6: AETDEDHAHWLEARM GenScript N/A Peptide 7: HAHWLEARMLLDNIY GenScript N/A Peptide 8: ARMLLDNIYLQDGLI GenScript N/A Peptide 9: NIYLQDGLIASLYRP GenScript N/A Peptide 10: GLIASLYRPEADKVA GenScript N/A Peptide 11: YRPEADKVAAIEGEF GenScript N/A (Continued on next page) e1 Cell Reports 35, 109107, May 11, 2021

Techniques: Control

Figure 3. ZIKV TCI-DNA vaccine protected female pregnant Ifnar1/ mice and their fetuses against ZIKV challenge Female Ifnar1/ mice were immunized with ZIKV E ectodomain, EDI/II peptides, TCI-DNA, or PBS control and mated with male Ifnar1/ mice for pregnancy at day 10 post-second immunization. The pregnant mice (E10–E12) were challenged with ZIKV (strain R103451, 104 PFU/mouse), and 6 days later, uteri and fetuses were collected to evaluate morphological changes as well as sera, body fluid, and tissues (including placenta) to measure viral titers using plaque-forming assay. Placenta was also observed for apoptosis and vascular damage and examined for inflammatory cytokines and chemokines as described below. (A and B) Representative image of morphology of uteri (E16–E18) and fetuses from pregnant mice challenged with ZIKV at E10-E12 (A), among which total numbers and dead fetuses from each group are shown (B). (C–E) Viral titers in placenta, fetal brain (C), as well as sera and amniotic fluid (D) and tissues (including brain, kidney, liver, spleen, heart, and muscle) (E), 6 days post-challenge. The detection limit was 12.5 PFU/g (for placenta and fetal brain), 20 PFU/g (for heart), 25 PFU/ml (for sera and amniotic fluid), and 25 PFU/g (for brain, kidney, liver, spleen, and muscle). The data are represented as mean ± SEM (n = 6). *p < 0.05; **p < 0.01; ***p < 0.001.

Journal: Cell reports

Article Title: A vaccine inducing solely cytotoxic T lymphocytes fully prevents Zika virus infection and fetal damage.

doi: 10.1016/j.celrep.2021.109107

Figure Lengend Snippet: Figure 3. ZIKV TCI-DNA vaccine protected female pregnant Ifnar1/ mice and their fetuses against ZIKV challenge Female Ifnar1/ mice were immunized with ZIKV E ectodomain, EDI/II peptides, TCI-DNA, or PBS control and mated with male Ifnar1/ mice for pregnancy at day 10 post-second immunization. The pregnant mice (E10–E12) were challenged with ZIKV (strain R103451, 104 PFU/mouse), and 6 days later, uteri and fetuses were collected to evaluate morphological changes as well as sera, body fluid, and tissues (including placenta) to measure viral titers using plaque-forming assay. Placenta was also observed for apoptosis and vascular damage and examined for inflammatory cytokines and chemokines as described below. (A and B) Representative image of morphology of uteri (E16–E18) and fetuses from pregnant mice challenged with ZIKV at E10-E12 (A), among which total numbers and dead fetuses from each group are shown (B). (C–E) Viral titers in placenta, fetal brain (C), as well as sera and amniotic fluid (D) and tissues (including brain, kidney, liver, spleen, heart, and muscle) (E), 6 days post-challenge. The detection limit was 12.5 PFU/g (for placenta and fetal brain), 20 PFU/g (for heart), 25 PFU/ml (for sera and amniotic fluid), and 25 PFU/g (for brain, kidney, liver, spleen, and muscle). The data are represented as mean ± SEM (n = 6). *p < 0.05; **p < 0.01; ***p < 0.001.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-mouse CD4 (IgG2b) antibody Bio X Cell BP0003-1; RRID:AB_1107636 Anti-mouse CD8a (IgG2b) antibody Bio X Cell BP0061; RRID:AB_1125541 Rat IgG2b isotype control antibody Bio X Cell BP0090; RRID:AB_1107780 Anti-mouse-CD8a (IgG2a) antibody Bio X Cell BE0004-1; RRID:AB_1107671 Rat IgG2a isotype control antibody Bio X Cell BE0089; RRID:AB_1107769 Anti-mouse IFNAR1 (MAR1-5A3) antibody Leinco I-401; RRID:AB_2491621 ZIKV EDIII-specific human antibody, ZV-64 Absolute Antibody Ab00811-10.0 Rabbit anti-vimentin antibody Abcam Ab92547; RRID:AB_10562134 Rabbit anti-active caspase-3 antibody Abcam Ab2302; RRID:AB_302962 FITC-anti-human antibody Sigma 9512-1mL; RRID:AB_259808 Anti-rabbit Alexa Fluor 647 antibody Abcam Ab150075; RRID:AB_2752244 FITC-anti-mouse antibody Biolegend 406001; RRID:AB_315029 Anti-flavivirus 4G2 antibody BEI Resources NR-50327 PerCP-Cy5.5-anti-mouse CD8 antibody BD Biosciences 551162; RRID:AB_394081 FITC-anti-mouse IL-2 antibody BD Biosciences 554427; RRID:AB_395385 FITC-anti-mouse CD4 antibody BD Biosciences 553729; RRID:AB_395013 PE-anti-mouse IFN-g antibody BD Biosciences 554412; RRID:AB_395376 Brilliant Violet 421-anti-mouse TNF-a antibody Biolegend 506328; RRID:AB_2562902 HRP-conjugated anti-mouse IgG-Fab antibody Sigma a9917-1mL; RRID:AB_258476 Anti-ZIKV NS3 antibody Genetex GTX133309; RRID:AB_2756864 Bacterial and virus strains Zika virus (human strain R103451 (2015/ Honduras)) BEI Resources NR-50355 Zika virus (human strain PAN2016 (2016/ Panama)) BEI Resources NR-50210 Chemicals, peptides, and recombinant proteins Zika virus full-length envelope (E) protein Aviva Systems Biology OPMA04848 Zika virus EDI/EDII peptides (see below peptide 1-5) BEI Resources NR-50553 Peptide 1: IGVSNRDFVEGMSGG BEI Resources N/A Peptide 2: TWVDVVLEHGGCVTV BEI Resources N/A Peptide 3: MAQDKPTVDIELVTT BEI Resources N/A Peptide 4: VDRGWGNGCGLFGKG BEI Resources N/A Peptide 5: VVLGSQEGAVHTALA BEI Resources N/A Zika virus NS3 overlapping peptides (see below peptide 6-15) GenScript N/A Peptide 6: AETDEDHAHWLEARM GenScript N/A Peptide 7: HAHWLEARMLLDNIY GenScript N/A Peptide 8: ARMLLDNIYLQDGLI GenScript N/A Peptide 9: NIYLQDGLIASLYRP GenScript N/A Peptide 10: GLIASLYRPEADKVA GenScript N/A Peptide 11: YRPEADKVAAIEGEF GenScript N/A (Continued on next page) e1 Cell Reports 35, 109107, May 11, 2021

Techniques: Control

Figure 4. ZIKV TCI-DNA vaccine prevented ZIKV-caused apoptosis in placenta of female pregnant Ifnar1/ mice Placenta collected from above ZIKV-challenged pregnant (E10–E12) Ifnar1/ mice was stained for activated form of caspase-3 (an apoptotic marker) by immunofluorescence assay. (A) Representative images of immunofluorescence staining of activated caspase-3 in placenta. ZIKV (green), activated caspase-3 (red), and nuclei (blue) were stained with anti-ZIKV antibody, anti-active caspase-3 antibody, and DAPI, respectively. The images were magnified at 633, with a scale bar of 10 mm. (B and C) Quantification of ZIKV+ (B) and activated caspase-3+ (C) staining in (A) by ImageJ software. The data are presented as mean ± SEM of fluorescence intensity for ZIKV+ or caspase-3+ staining in each field (n = 6; n indicates numbers of images from different placentas). *p < 0.05; **p < 0.01; ***p < 0.001.

Journal: Cell reports

Article Title: A vaccine inducing solely cytotoxic T lymphocytes fully prevents Zika virus infection and fetal damage.

doi: 10.1016/j.celrep.2021.109107

Figure Lengend Snippet: Figure 4. ZIKV TCI-DNA vaccine prevented ZIKV-caused apoptosis in placenta of female pregnant Ifnar1/ mice Placenta collected from above ZIKV-challenged pregnant (E10–E12) Ifnar1/ mice was stained for activated form of caspase-3 (an apoptotic marker) by immunofluorescence assay. (A) Representative images of immunofluorescence staining of activated caspase-3 in placenta. ZIKV (green), activated caspase-3 (red), and nuclei (blue) were stained with anti-ZIKV antibody, anti-active caspase-3 antibody, and DAPI, respectively. The images were magnified at 633, with a scale bar of 10 mm. (B and C) Quantification of ZIKV+ (B) and activated caspase-3+ (C) staining in (A) by ImageJ software. The data are presented as mean ± SEM of fluorescence intensity for ZIKV+ or caspase-3+ staining in each field (n = 6; n indicates numbers of images from different placentas). *p < 0.05; **p < 0.01; ***p < 0.001.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-mouse CD4 (IgG2b) antibody Bio X Cell BP0003-1; RRID:AB_1107636 Anti-mouse CD8a (IgG2b) antibody Bio X Cell BP0061; RRID:AB_1125541 Rat IgG2b isotype control antibody Bio X Cell BP0090; RRID:AB_1107780 Anti-mouse-CD8a (IgG2a) antibody Bio X Cell BE0004-1; RRID:AB_1107671 Rat IgG2a isotype control antibody Bio X Cell BE0089; RRID:AB_1107769 Anti-mouse IFNAR1 (MAR1-5A3) antibody Leinco I-401; RRID:AB_2491621 ZIKV EDIII-specific human antibody, ZV-64 Absolute Antibody Ab00811-10.0 Rabbit anti-vimentin antibody Abcam Ab92547; RRID:AB_10562134 Rabbit anti-active caspase-3 antibody Abcam Ab2302; RRID:AB_302962 FITC-anti-human antibody Sigma 9512-1mL; RRID:AB_259808 Anti-rabbit Alexa Fluor 647 antibody Abcam Ab150075; RRID:AB_2752244 FITC-anti-mouse antibody Biolegend 406001; RRID:AB_315029 Anti-flavivirus 4G2 antibody BEI Resources NR-50327 PerCP-Cy5.5-anti-mouse CD8 antibody BD Biosciences 551162; RRID:AB_394081 FITC-anti-mouse IL-2 antibody BD Biosciences 554427; RRID:AB_395385 FITC-anti-mouse CD4 antibody BD Biosciences 553729; RRID:AB_395013 PE-anti-mouse IFN-g antibody BD Biosciences 554412; RRID:AB_395376 Brilliant Violet 421-anti-mouse TNF-a antibody Biolegend 506328; RRID:AB_2562902 HRP-conjugated anti-mouse IgG-Fab antibody Sigma a9917-1mL; RRID:AB_258476 Anti-ZIKV NS3 antibody Genetex GTX133309; RRID:AB_2756864 Bacterial and virus strains Zika virus (human strain R103451 (2015/ Honduras)) BEI Resources NR-50355 Zika virus (human strain PAN2016 (2016/ Panama)) BEI Resources NR-50210 Chemicals, peptides, and recombinant proteins Zika virus full-length envelope (E) protein Aviva Systems Biology OPMA04848 Zika virus EDI/EDII peptides (see below peptide 1-5) BEI Resources NR-50553 Peptide 1: IGVSNRDFVEGMSGG BEI Resources N/A Peptide 2: TWVDVVLEHGGCVTV BEI Resources N/A Peptide 3: MAQDKPTVDIELVTT BEI Resources N/A Peptide 4: VDRGWGNGCGLFGKG BEI Resources N/A Peptide 5: VVLGSQEGAVHTALA BEI Resources N/A Zika virus NS3 overlapping peptides (see below peptide 6-15) GenScript N/A Peptide 6: AETDEDHAHWLEARM GenScript N/A Peptide 7: HAHWLEARMLLDNIY GenScript N/A Peptide 8: ARMLLDNIYLQDGLI GenScript N/A Peptide 9: NIYLQDGLIASLYRP GenScript N/A Peptide 10: GLIASLYRPEADKVA GenScript N/A Peptide 11: YRPEADKVAAIEGEF GenScript N/A (Continued on next page) e1 Cell Reports 35, 109107, May 11, 2021

Techniques: Staining, Marker, Software

Figure 5. ZIKV TCI-DNA vaccine prevented ZIKV-caused inflammation in placenta of female pregnant Ifnar1/ mice Placentas collected from above challenged pregnant (E10–E12) Ifnar1/ mice were examined for inflammatory cytokines (A) and chemokines (B) by Mouse Inflammatory Cytokines Multi-Analyte ELISArray Kit and Mouse Common Chemokines Multi-Analyte ELISArray Kit, respectively. The data are presented as mean ± SEM (n = 6). *p < 0.05; **p < 0.01; ***p < 0.001.

Journal: Cell reports

Article Title: A vaccine inducing solely cytotoxic T lymphocytes fully prevents Zika virus infection and fetal damage.

doi: 10.1016/j.celrep.2021.109107

Figure Lengend Snippet: Figure 5. ZIKV TCI-DNA vaccine prevented ZIKV-caused inflammation in placenta of female pregnant Ifnar1/ mice Placentas collected from above challenged pregnant (E10–E12) Ifnar1/ mice were examined for inflammatory cytokines (A) and chemokines (B) by Mouse Inflammatory Cytokines Multi-Analyte ELISArray Kit and Mouse Common Chemokines Multi-Analyte ELISArray Kit, respectively. The data are presented as mean ± SEM (n = 6). *p < 0.05; **p < 0.01; ***p < 0.001.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-mouse CD4 (IgG2b) antibody Bio X Cell BP0003-1; RRID:AB_1107636 Anti-mouse CD8a (IgG2b) antibody Bio X Cell BP0061; RRID:AB_1125541 Rat IgG2b isotype control antibody Bio X Cell BP0090; RRID:AB_1107780 Anti-mouse-CD8a (IgG2a) antibody Bio X Cell BE0004-1; RRID:AB_1107671 Rat IgG2a isotype control antibody Bio X Cell BE0089; RRID:AB_1107769 Anti-mouse IFNAR1 (MAR1-5A3) antibody Leinco I-401; RRID:AB_2491621 ZIKV EDIII-specific human antibody, ZV-64 Absolute Antibody Ab00811-10.0 Rabbit anti-vimentin antibody Abcam Ab92547; RRID:AB_10562134 Rabbit anti-active caspase-3 antibody Abcam Ab2302; RRID:AB_302962 FITC-anti-human antibody Sigma 9512-1mL; RRID:AB_259808 Anti-rabbit Alexa Fluor 647 antibody Abcam Ab150075; RRID:AB_2752244 FITC-anti-mouse antibody Biolegend 406001; RRID:AB_315029 Anti-flavivirus 4G2 antibody BEI Resources NR-50327 PerCP-Cy5.5-anti-mouse CD8 antibody BD Biosciences 551162; RRID:AB_394081 FITC-anti-mouse IL-2 antibody BD Biosciences 554427; RRID:AB_395385 FITC-anti-mouse CD4 antibody BD Biosciences 553729; RRID:AB_395013 PE-anti-mouse IFN-g antibody BD Biosciences 554412; RRID:AB_395376 Brilliant Violet 421-anti-mouse TNF-a antibody Biolegend 506328; RRID:AB_2562902 HRP-conjugated anti-mouse IgG-Fab antibody Sigma a9917-1mL; RRID:AB_258476 Anti-ZIKV NS3 antibody Genetex GTX133309; RRID:AB_2756864 Bacterial and virus strains Zika virus (human strain R103451 (2015/ Honduras)) BEI Resources NR-50355 Zika virus (human strain PAN2016 (2016/ Panama)) BEI Resources NR-50210 Chemicals, peptides, and recombinant proteins Zika virus full-length envelope (E) protein Aviva Systems Biology OPMA04848 Zika virus EDI/EDII peptides (see below peptide 1-5) BEI Resources NR-50553 Peptide 1: IGVSNRDFVEGMSGG BEI Resources N/A Peptide 2: TWVDVVLEHGGCVTV BEI Resources N/A Peptide 3: MAQDKPTVDIELVTT BEI Resources N/A Peptide 4: VDRGWGNGCGLFGKG BEI Resources N/A Peptide 5: VVLGSQEGAVHTALA BEI Resources N/A Zika virus NS3 overlapping peptides (see below peptide 6-15) GenScript N/A Peptide 6: AETDEDHAHWLEARM GenScript N/A Peptide 7: HAHWLEARMLLDNIY GenScript N/A Peptide 8: ARMLLDNIYLQDGLI GenScript N/A Peptide 9: NIYLQDGLIASLYRP GenScript N/A Peptide 10: GLIASLYRPEADKVA GenScript N/A Peptide 11: YRPEADKVAAIEGEF GenScript N/A (Continued on next page) e1 Cell Reports 35, 109107, May 11, 2021

Techniques:

Figure 6. ZIKV TCI-DNA vaccine induced low to no ZIKV-, E-, and NS3-specific antibodies without ADE effect Mouse sera collected at 10 days post-second immunization were detected for ZIKV E-, NS3-, and ZIKV-specific IgG antibody, neutralizing antibodies, and ADE of ZIKV infection. ZIKV strain R103451 was used for the neutralization and ADE tests. (A and B) ELISA was used for detection of IgG antibody specific to ZIKV E ectodomain, NS3 peptides, and ZIKV (R103451 strain) in sera of BALB/c (A) or Ifnar1/

Journal: Cell reports

Article Title: A vaccine inducing solely cytotoxic T lymphocytes fully prevents Zika virus infection and fetal damage.

doi: 10.1016/j.celrep.2021.109107

Figure Lengend Snippet: Figure 6. ZIKV TCI-DNA vaccine induced low to no ZIKV-, E-, and NS3-specific antibodies without ADE effect Mouse sera collected at 10 days post-second immunization were detected for ZIKV E-, NS3-, and ZIKV-specific IgG antibody, neutralizing antibodies, and ADE of ZIKV infection. ZIKV strain R103451 was used for the neutralization and ADE tests. (A and B) ELISA was used for detection of IgG antibody specific to ZIKV E ectodomain, NS3 peptides, and ZIKV (R103451 strain) in sera of BALB/c (A) or Ifnar1/

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-mouse CD4 (IgG2b) antibody Bio X Cell BP0003-1; RRID:AB_1107636 Anti-mouse CD8a (IgG2b) antibody Bio X Cell BP0061; RRID:AB_1125541 Rat IgG2b isotype control antibody Bio X Cell BP0090; RRID:AB_1107780 Anti-mouse-CD8a (IgG2a) antibody Bio X Cell BE0004-1; RRID:AB_1107671 Rat IgG2a isotype control antibody Bio X Cell BE0089; RRID:AB_1107769 Anti-mouse IFNAR1 (MAR1-5A3) antibody Leinco I-401; RRID:AB_2491621 ZIKV EDIII-specific human antibody, ZV-64 Absolute Antibody Ab00811-10.0 Rabbit anti-vimentin antibody Abcam Ab92547; RRID:AB_10562134 Rabbit anti-active caspase-3 antibody Abcam Ab2302; RRID:AB_302962 FITC-anti-human antibody Sigma 9512-1mL; RRID:AB_259808 Anti-rabbit Alexa Fluor 647 antibody Abcam Ab150075; RRID:AB_2752244 FITC-anti-mouse antibody Biolegend 406001; RRID:AB_315029 Anti-flavivirus 4G2 antibody BEI Resources NR-50327 PerCP-Cy5.5-anti-mouse CD8 antibody BD Biosciences 551162; RRID:AB_394081 FITC-anti-mouse IL-2 antibody BD Biosciences 554427; RRID:AB_395385 FITC-anti-mouse CD4 antibody BD Biosciences 553729; RRID:AB_395013 PE-anti-mouse IFN-g antibody BD Biosciences 554412; RRID:AB_395376 Brilliant Violet 421-anti-mouse TNF-a antibody Biolegend 506328; RRID:AB_2562902 HRP-conjugated anti-mouse IgG-Fab antibody Sigma a9917-1mL; RRID:AB_258476 Anti-ZIKV NS3 antibody Genetex GTX133309; RRID:AB_2756864 Bacterial and virus strains Zika virus (human strain R103451 (2015/ Honduras)) BEI Resources NR-50355 Zika virus (human strain PAN2016 (2016/ Panama)) BEI Resources NR-50210 Chemicals, peptides, and recombinant proteins Zika virus full-length envelope (E) protein Aviva Systems Biology OPMA04848 Zika virus EDI/EDII peptides (see below peptide 1-5) BEI Resources NR-50553 Peptide 1: IGVSNRDFVEGMSGG BEI Resources N/A Peptide 2: TWVDVVLEHGGCVTV BEI Resources N/A Peptide 3: MAQDKPTVDIELVTT BEI Resources N/A Peptide 4: VDRGWGNGCGLFGKG BEI Resources N/A Peptide 5: VVLGSQEGAVHTALA BEI Resources N/A Zika virus NS3 overlapping peptides (see below peptide 6-15) GenScript N/A Peptide 6: AETDEDHAHWLEARM GenScript N/A Peptide 7: HAHWLEARMLLDNIY GenScript N/A Peptide 8: ARMLLDNIYLQDGLI GenScript N/A Peptide 9: NIYLQDGLIASLYRP GenScript N/A Peptide 10: GLIASLYRPEADKVA GenScript N/A Peptide 11: YRPEADKVAAIEGEF GenScript N/A (Continued on next page) e1 Cell Reports 35, 109107, May 11, 2021

Techniques: Infection, Neutralization, Enzyme-linked Immunosorbent Assay

Figure 7. CD8+ T cell immune responses induced by ZIKV TCI-DNA vaccine played a key role in protecting pregnant mothers and their fe- tuses against ZIKV infection Female BALB/c mice were immunized with ZIKV TCI-DNA or PBS control for two doses and mated with male BALB/c mice for pregnancy at 10 days post-second immunization. The pregnant (E10–E12) mice were then injected (i.p.) with anti-CD8a (for depleting CD8+ T cells) or IgG2a isotype control (i.e., Iso con; without depleting CD8+ T cells) antibody (200 mg/mouse) for three times (2, 1, and 3 days p.i.). One day before challenge, the mice were also injected with anti-IFNAR1 blocking antibody (for depleting type I IFN; 2 mg/mouse) and then infected with ZIKV (strain R103451, 106 PFU/mouse). (A–C) Six days post-challenge, the mice were euthanized, recorded for morphology of uteri and fetuses (A), and counted for dead and total fetuses (B), and viral titers were determined by plaque-forming assay in placenta, amniotic fluid, and fetal brain, as well as sera collected at 3 and 6 days post-challenge (C). The detection limit was 20 PFU/g (for placenta), 40 PFU/g (for fetal brain), and 50 PFU/ml (for sera and amniotic fluid). Six days post-challenge, splenocytes were isolated from the mice injected with isotype control antibody (i.e., Iso con) and analyzed for ZIKV-specific CD8+ T cell responses by flow cytometry analysis. (D) Quantification of the frequencies of CD8+ T cells (left), as well as IL2+, IFN-g+, and tumor necrosis factor (TNF)-a secretion in CD8+ T cells (right) in splenocytes. The data are represented as mean ± SEM (n = 6). *p < 0.05; **p < 0.01; ***p < 0.001.

Journal: Cell reports

Article Title: A vaccine inducing solely cytotoxic T lymphocytes fully prevents Zika virus infection and fetal damage.

doi: 10.1016/j.celrep.2021.109107

Figure Lengend Snippet: Figure 7. CD8+ T cell immune responses induced by ZIKV TCI-DNA vaccine played a key role in protecting pregnant mothers and their fe- tuses against ZIKV infection Female BALB/c mice were immunized with ZIKV TCI-DNA or PBS control for two doses and mated with male BALB/c mice for pregnancy at 10 days post-second immunization. The pregnant (E10–E12) mice were then injected (i.p.) with anti-CD8a (for depleting CD8+ T cells) or IgG2a isotype control (i.e., Iso con; without depleting CD8+ T cells) antibody (200 mg/mouse) for three times (2, 1, and 3 days p.i.). One day before challenge, the mice were also injected with anti-IFNAR1 blocking antibody (for depleting type I IFN; 2 mg/mouse) and then infected with ZIKV (strain R103451, 106 PFU/mouse). (A–C) Six days post-challenge, the mice were euthanized, recorded for morphology of uteri and fetuses (A), and counted for dead and total fetuses (B), and viral titers were determined by plaque-forming assay in placenta, amniotic fluid, and fetal brain, as well as sera collected at 3 and 6 days post-challenge (C). The detection limit was 20 PFU/g (for placenta), 40 PFU/g (for fetal brain), and 50 PFU/ml (for sera and amniotic fluid). Six days post-challenge, splenocytes were isolated from the mice injected with isotype control antibody (i.e., Iso con) and analyzed for ZIKV-specific CD8+ T cell responses by flow cytometry analysis. (D) Quantification of the frequencies of CD8+ T cells (left), as well as IL2+, IFN-g+, and tumor necrosis factor (TNF)-a secretion in CD8+ T cells (right) in splenocytes. The data are represented as mean ± SEM (n = 6). *p < 0.05; **p < 0.01; ***p < 0.001.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-mouse CD4 (IgG2b) antibody Bio X Cell BP0003-1; RRID:AB_1107636 Anti-mouse CD8a (IgG2b) antibody Bio X Cell BP0061; RRID:AB_1125541 Rat IgG2b isotype control antibody Bio X Cell BP0090; RRID:AB_1107780 Anti-mouse-CD8a (IgG2a) antibody Bio X Cell BE0004-1; RRID:AB_1107671 Rat IgG2a isotype control antibody Bio X Cell BE0089; RRID:AB_1107769 Anti-mouse IFNAR1 (MAR1-5A3) antibody Leinco I-401; RRID:AB_2491621 ZIKV EDIII-specific human antibody, ZV-64 Absolute Antibody Ab00811-10.0 Rabbit anti-vimentin antibody Abcam Ab92547; RRID:AB_10562134 Rabbit anti-active caspase-3 antibody Abcam Ab2302; RRID:AB_302962 FITC-anti-human antibody Sigma 9512-1mL; RRID:AB_259808 Anti-rabbit Alexa Fluor 647 antibody Abcam Ab150075; RRID:AB_2752244 FITC-anti-mouse antibody Biolegend 406001; RRID:AB_315029 Anti-flavivirus 4G2 antibody BEI Resources NR-50327 PerCP-Cy5.5-anti-mouse CD8 antibody BD Biosciences 551162; RRID:AB_394081 FITC-anti-mouse IL-2 antibody BD Biosciences 554427; RRID:AB_395385 FITC-anti-mouse CD4 antibody BD Biosciences 553729; RRID:AB_395013 PE-anti-mouse IFN-g antibody BD Biosciences 554412; RRID:AB_395376 Brilliant Violet 421-anti-mouse TNF-a antibody Biolegend 506328; RRID:AB_2562902 HRP-conjugated anti-mouse IgG-Fab antibody Sigma a9917-1mL; RRID:AB_258476 Anti-ZIKV NS3 antibody Genetex GTX133309; RRID:AB_2756864 Bacterial and virus strains Zika virus (human strain R103451 (2015/ Honduras)) BEI Resources NR-50355 Zika virus (human strain PAN2016 (2016/ Panama)) BEI Resources NR-50210 Chemicals, peptides, and recombinant proteins Zika virus full-length envelope (E) protein Aviva Systems Biology OPMA04848 Zika virus EDI/EDII peptides (see below peptide 1-5) BEI Resources NR-50553 Peptide 1: IGVSNRDFVEGMSGG BEI Resources N/A Peptide 2: TWVDVVLEHGGCVTV BEI Resources N/A Peptide 3: MAQDKPTVDIELVTT BEI Resources N/A Peptide 4: VDRGWGNGCGLFGKG BEI Resources N/A Peptide 5: VVLGSQEGAVHTALA BEI Resources N/A Zika virus NS3 overlapping peptides (see below peptide 6-15) GenScript N/A Peptide 6: AETDEDHAHWLEARM GenScript N/A Peptide 7: HAHWLEARMLLDNIY GenScript N/A Peptide 8: ARMLLDNIYLQDGLI GenScript N/A Peptide 9: NIYLQDGLIASLYRP GenScript N/A Peptide 10: GLIASLYRPEADKVA GenScript N/A Peptide 11: YRPEADKVAAIEGEF GenScript N/A (Continued on next page) e1 Cell Reports 35, 109107, May 11, 2021

Techniques: Infection, Control, Injection, Blocking Assay, Isolation, Cytometry