Journal: Viruses

Article Title: Evaluation of a Lyophilized CRISPR-Cas12 Assay for a Sensitive, Specific, and Rapid Detection of SARS-CoV-2

doi: 10.3390/v13030420

Figure Lengend Snippet: Cross-reaction evaluation of Lyo-CRISPR SARS-CoV-2 kit with other respiratory pathogens.

Article Snippet: Respiratory syncytial virus , ATCC ® VR-1580DQ , 10 5 copies/mL , 0/3 , Negative , 3/3 , Valid.

Techniques: Concentration Assay

Journal: Molecular therapy : the journal of the American Society of Gene Therapy

Article Title: Neuraminidase delivered as an APC-targeted DNA vaccine induces protective antibodies against influenza.

doi: 10.1016/j.ymthe.2023.03.012

Figure Lengend Snippet: Figure 1. In vitro characterization of MHC class II-targeted heterodimeric DNA vaccines containing NA and/or HA (A) MHC class II-targeted heterodimeric acid/base (A/B) vaccine proteins consist of A and B chains, each having a targeting, dimerization, and antigenic unit. The targeting unit is either an scFv that binds MHC class II on APCs (scFvaMHCII, green) or a non-targeted control that binds the hapten NIP (scFvaNIP, white). The targeting units are genetically linked to a shortened hinge from human IgG3 (exon h1), allowing covalent disulfide bond formation between the A and B chains (black lines). The hinge is followed by a duplicated A (red) or B (blue) heterodimerization unit based on a modified Jun/Fos leucine zipper motif enriched for acidic or basic amino acids, respectively. The A and B chains are linked to antigenic units composed of NA (dark blue) or HA (orange) derived from influenza H1N1 PR8 virus. The heterodimeric vaccine proteins are named as

Article Snippet: The copy number of viral RNA was determined with quantitative genomic RNA from H1N1 PR8 (VR-95DQ, ATCC) and shown per microgram total tissue RNA.

Techniques: In Vitro, Vaccines, Control, Derivative Assay, Virus

Journal: Molecular therapy : the journal of the American Society of Gene Therapy

Article Title: Neuraminidase delivered as an APC-targeted DNA vaccine induces protective antibodies against influenza.

doi: 10.1016/j.ymthe.2023.03.012

Figure Lengend Snippet: Figure 2. MHC class II-targeted NA-bivalent heterodimers efficiently induce antibody responses after a single DNA vaccination (A) BALB/c mice were vaccinated once i.m. with 5 mg of each A and B plasmids (10 mg DNA in total) encoding the indicated heterodimeric vaccine proteins (box), followed immediately by EP. Blood was sampled at the indicated time points. (B) NA-specific (top) and HA-specific (bottom) IgG (left), IgG1 (center), and IgG2a (right) titers were measured by ELISA at the indicated time points after immunization. Statistical analysis compared targeted vs. non-targeted and antigen-bivalent vs. monovalent groups. (C) BALB/c mice were immunized once i.m./EP with different amounts of the indicated DNA vaccines. Serum was analyzed for NA-specific IgG responses 4 weeks post vaccination. (D) IgG responses specific for inactivated H1N1 PR8 virus (left), rec. NA protein (center), or rec. HA protein (right) in sera harvested 6 weeks post vaccination from

Article Snippet: The copy number of viral RNA was determined with quantitative genomic RNA from H1N1 PR8 (VR-95DQ, ATCC) and shown per microgram total tissue RNA.

Techniques: Enzyme-linked Immunosorbent Assay, Vaccines, Virus

Journal: Molecular therapy : the journal of the American Society of Gene Therapy

Article Title: Neuraminidase delivered as an APC-targeted DNA vaccine induces protective antibodies against influenza.

doi: 10.1016/j.ymthe.2023.03.012

Figure Lengend Snippet: Figure 4. A single DNA immunization with MHC class II-targeted NA/NA vaccines protects against homologous influenza virus challenge (A) BALB/c mice were immunized once i.m./EP and challenged with homologous influenza A H1N1 PR8 virus at week 2 (short term) or between weeks 6 and 10 (long term) after vaccination. (B–F) 50 mg of each A and B plasmids (total 100 mg, B), 5 mg/ plasmid (10 mg total, C–E), or 1 mg/plasmid (2 mg total, F) of the indicated DNA vaccines (box) were used. Mice were challenged with a lethal dose of 5 LD50 (B–D and F) or 100 LD50 (E) at week 9 (B), week 10 (C), week 2 (D and E), or week 6 (F). Weight (left, mean ± SEM) and survival (right) were monitored up to 12 days post challenge. n = 8 mice/group (B, C, E, and F) or n = 6 mice/group (D). Dead mice were assigned a weight of 75%. *p % 0.05, ***p % 0.001; two-way ANOVA (weight curve) or Mantel-Cox test (survival). Statistical analysis compared MHC class II- targeted NA/NA versus HA/HA as well as targeted versus non-targeted and anti- gen-bivalent versus -monovalent NA.

Article Snippet: The copy number of viral RNA was determined with quantitative genomic RNA from H1N1 PR8 (VR-95DQ, ATCC) and shown per microgram total tissue RNA.

Techniques: Vaccines, Virus, Plasmid Preparation

Journal: Molecular therapy : the journal of the American Society of Gene Therapy

Article Title: Neuraminidase delivered as an APC-targeted DNA vaccine induces protective antibodies against influenza.

doi: 10.1016/j.ymthe.2023.03.012

Figure Lengend Snippet: Figure 5. MHC class II-targeted NA DNA vaccine induces superior NA-specific antibody responses compared with conventional inactivated virus after one vaccination (A, B) BALB/c mice were immunized once i.m. with 0.2, 1, or 5 mg of formalin-inactivated H1N1 PR8 virus or with 5 mg/ plasmid (10 mg total) of the indicated DNA vaccines, fol- lowed by EP (box). (A) NA-specific (left) and HA-specific (right) titers were measured by ELISA at the indicated time points after immunization. Mean ± SEM is shown. (B) Mice were challenged with 5 LD50 H1N1 PR8 virus 11 week post vaccination. Weight (left, mean ± SEM) and survival (right) were monitored up to 12 days post challenge. Dead mice were assigned a weight of 75%; n = 8 mice/group. Statistical analysis compared the different doses of inactivated virus with DNA vaccination with MHC class II-targeted NA and/or HA. **p % 0.01, ***p % 0.001; two-way ANOVA (IgG titers, weight curve) or Mantel-Cox test (survival).

Article Snippet: The copy number of viral RNA was determined with quantitative genomic RNA from H1N1 PR8 (VR-95DQ, ATCC) and shown per microgram total tissue RNA.

Techniques: Virus, Plasmid Preparation, Vaccines, Enzyme-linked Immunosorbent Assay

Journal: Molecular therapy : the journal of the American Society of Gene Therapy

Article Title: Neuraminidase delivered as an APC-targeted DNA vaccine induces protective antibodies against influenza.

doi: 10.1016/j.ymthe.2023.03.012

Figure Lengend Snippet: Figure 6. Vaccine-induced NA-specific antibodies mediate protection against homologous influenza infection, inhibit NA enzymatic activity, and block viral replication (A–D) BALB/c mice were immunized once i.m./EP with 50 mg each of A and B plasmids (100 mg total) as indicated. (A) Pooled sera (350 mL) from mice 5 weeks after vaccination were transferred i.p. to naive BALB/c mice. One day after transfer, mice were challenged with 2.5 LD50 H1N1 PR8 virus. (B) 2 weeks after vaccination with the indicated DNA vaccines, mice were challenged with 5 LD50 H1N1 PR8 virus. Starting on day 12 after vaccination, mice were treated with anti-CD4 and anti- CD8 mAbs or isotype-matched control mAbs every second day. n = 8 mice/group; weight (left, mean ± SEM) and survival (right) were monitored up to 12 days post challenge, and dead mice were assigned a weight of 75% (A and B). (C) Neutrali- zation of influenza A H1N1 PR8 virus by serum from vaccinated mice (box) obtained 4 weeks after immunization was measured by microneutralization assay and is shown as IC50. (D) The same sera were tested for ability to inhibit NA enzyme activity by ELLA, using a reassorted influenza A H6N1 virus containing NA from H1N1 PR8. IC50 is indicated. (E and F) BALB/c mice were immunized once i.m./EP with 50 mg of each A and B plasmids (100 mg total, E) or 5 mg of each A and B plasmids (10 mg total, F) as indicated. Mice were challenged with homologous H1N1 PR8 virus 6 weeks after vaccination, and lungs were harvested on day 5 post challenge. Viral loads were determined by qRT-PCR (left), showing gene copy number per micro- gram total tissue RNA, and by measuring in vitro cell culture infectivity (right), pre- sented as 50% tissue culture infectious dose (TCID50) per milliliter of lung homog- enates. Individual mice and mean ± SEM are shown; n = 4–6 mice/group (C–F). *p % 0.05, **p % 0.01, ***p % 0.001; two-way ANOVA (weight curve), Mantel-Cox test (survival), Kruskal-Wallis multiple-comparisons test with Dunn’s correction (C and D, TCID50), or multiple-comparisons one-way ANOVA with Tukey’s correction (copy number).

Article Snippet: The copy number of viral RNA was determined with quantitative genomic RNA from H1N1 PR8 (VR-95DQ, ATCC) and shown per microgram total tissue RNA.

Techniques: Infection, Activity Assay, Blocking Assay, Virus, Vaccines, Control, Microneutralization Assay, Quantitative RT-PCR, In Vitro, Cell Culture