Journal: Vaccines

Article Title: Immunogenicity and Efficacy of a Trivalent HSV-2 gC2, gD2, gE2 Nucleoside-Modified mRNA-LNP Vaccine Against HSV-1 Eye Infection and Neuroinvasion in Mice

doi: 10.3390/vaccines14030253

Figure Lengend Snippet: Inoculation of HSV-1 produces dose-dependent changes in survival, blepharoconjunctivitis, and weight loss. HSV-1 was inoculated onto the intact cornea of each eye. ( A ) Survival curve. p values calculated by the Log-rank test. n = 5 mice/group ( B ) The blepharoconjunctivitis disease score is shown for the most affected eye on the most severe day of disease. n = 5 mice/group. ( C ) Percent weight loss at time of humane euthanasia or at the end of the study on day 14 post-infection. n = 5 mice/group, except n = 4 for the 10 4 group. p values for ( B , C ) were calculated by Ordinary one-way ANOVA and Dunnett’s multiple comparisons test. * p < 0.05, ** p < 0.01, *** p < 0.001; **** p < 0.0001; ns, not significant.

Article Snippet: To determine HSV-1 DNA copy number, we ran purified HSV-1 DNA (ATCC, VR-539DQ) or adipsin DNA (BioChain, Princeton, NJ, USA, D1334149) as standards.

Techniques: Infection

Journal: Vaccines

Article Title: Immunogenicity and Efficacy of a Trivalent HSV-2 gC2, gD2, gE2 Nucleoside-Modified mRNA-LNP Vaccine Against HSV-1 Eye Infection and Neuroinvasion in Mice

doi: 10.3390/vaccines14030253

Figure Lengend Snippet: Kinetics of infection after HSV-1 was placed on the intact cornea of each eye. Animals were infected with 10 5 PFU (1 lethal dose 50, LD 50 ) of HSV-1 in each eye and sacrificed on days 1, 3, 5, and 7 post-infection ( n = 5 per group). Each tissue was processed separately and analyzed for virus titers (blue) and HSV-1 DNA copy number by qPCR (red). The tissues included ( A ) eye (eye bulb and attached ocular conjunctiva), ( B ) trigeminal ganglion, ( C ) brainstem, ( D ) olfactory bulb, and ( E ) cerebrum/cerebellum.

Article Snippet: To determine HSV-1 DNA copy number, we ran purified HSV-1 DNA (ATCC, VR-539DQ) or adipsin DNA (BioChain, Princeton, NJ, USA, D1334149) as standards.

Techniques: Infection, Virus, Olfactory

Journal: Vaccines

Article Title: Immunogenicity and Efficacy of a Trivalent HSV-2 gC2, gD2, gE2 Nucleoside-Modified mRNA-LNP Vaccine Against HSV-1 Eye Infection and Neuroinvasion in Mice

doi: 10.3390/vaccines14030253

Figure Lengend Snippet: Serum IgG ELISA (binding) and neutralizing antibodies. Mice were immunized twice IM with PBS or 10 μg of the trivalent HSV-2 mRNA vaccine. Sera were obtained 4 weeks after the second immunization, before infection with HSV-1. ( A ) Serum IgG ELISA endpoint titers against HSV-2 or cross-reacting antibodies against HSV-1. ( B ) Neutralizing antibodies to HSV-1. n = 9 or 10 animals/group. p values were calculated using the two-tailed Mann–Whitney test. **** p < 0.0001; ns, not significant.

Article Snippet: To determine HSV-1 DNA copy number, we ran purified HSV-1 DNA (ATCC, VR-539DQ) or adipsin DNA (BioChain, Princeton, NJ, USA, D1334149) as standards.

Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay, Infection, Two Tailed Test, MANN-WHITNEY

Journal: Vaccines

Article Title: Immunogenicity and Efficacy of a Trivalent HSV-2 gC2, gD2, gE2 Nucleoside-Modified mRNA-LNP Vaccine Against HSV-1 Eye Infection and Neuroinvasion in Mice

doi: 10.3390/vaccines14030253

Figure Lengend Snippet: Vaccine protection against disease after HSV-1 inoculation on the intact cornea of each eye. Mice were immunized with PBS or 10 μg trivalent mRNA, and 28 days after the second immunization, the intact cornea of each eye was inoculated with 10 6 PFU (10 LD 50 ) HSV-1 ( n = 10/group). ( A ) Survival curve. The p value was determined using the Log-rank test. ( B ) Blepharoconjunctivitis disease score on the most severe day of disease post-infection for each animal. ( C ) Weight loss. p values for ( B , C ) were calculated using the two-tailed Mann–Whitney test. n = 10/group; ** p < 0.01; *** p < 0.001.

Article Snippet: To determine HSV-1 DNA copy number, we ran purified HSV-1 DNA (ATCC, VR-539DQ) or adipsin DNA (BioChain, Princeton, NJ, USA, D1334149) as standards.

Techniques: Infection, Two Tailed Test, MANN-WHITNEY

Journal: Vaccines

Article Title: Immunogenicity and Efficacy of a Trivalent HSV-2 gC2, gD2, gE2 Nucleoside-Modified mRNA-LNP Vaccine Against HSV-1 Eye Infection and Neuroinvasion in Mice

doi: 10.3390/vaccines14030253

Figure Lengend Snippet: Vaccine protection as assessed by virus culture and HSV-1 DNA copy number in tissues after HSV-1 inoculation of the intact cornea of each eye. Mice were immunized twice with PBS or 10 μg of the trivalent HSV-2 mRNA vaccine, and challenged in each eye with 10 6 plaque forming units (PFU) HSV-1 (10 LD 50 ). ( A ) Tissues were harvested 5 days post-infection for viral titers in PBS and vaccine groups ( n = 5/group). ( B ) The same tissues were evaluated for HSV-1 DNA copy number. ( C ) Tissues were harvested 7 weeks post-infection for HSV-1 DNA copy number in mice in the vaccine group ( n = 10). The black symbol indicates the same mouse with HSV-1 DNA in the eye and the trigeminal ganglia. p values were calculated by the two-tailed Mann–Whitney test; * p < 0.05; ** p < 0.01; ns, not significant.

Article Snippet: To determine HSV-1 DNA copy number, we ran purified HSV-1 DNA (ATCC, VR-539DQ) or adipsin DNA (BioChain, Princeton, NJ, USA, D1334149) as standards.

Techniques: Virus, Infection, Two Tailed Test, MANN-WHITNEY

Journal: Vaccines

Article Title: Immunogenicity and Efficacy of a Trivalent HSV-2 gC2, gD2, gE2 Nucleoside-Modified mRNA-LNP Vaccine Against HSV-1 Eye Infection and Neuroinvasion in Mice

doi: 10.3390/vaccines14030253

Figure Lengend Snippet: Representative images of the eyelids and palpebral conjunctivae, and the trigeminal ganglia of mice infected with HSV-1 that were unvaccinated or vaccinated with the trivalent HSV-2 mRNA vaccine. ( a , b ) Evaluation for blepharoconjunctivitis in an immunized animal: The eyelid shows minimal mixed inflammatory cell infiltrates in the conjunctival substantia propria (arrow). No HSV-1 immunolabelling is detected by IHC. H&E ( a ) and IHC for HSV-1 ( b ). ( c , d ) Evaluation for blepharoconjunctivitis in an unimmunized mouse: Blepharoconjunctivitis is visible (arrow) with regional necrosis of the epithelium (asterisk) and strong immunolabeling of the surface and follicular epithelium for HSV-1. H&E ( c ) and IHC for HSV1 ( d ). Scale bar for ( a – d ), 100 µm. ( e , f ) Trigeminal ganglion of an immunized mouse: The trigeminal ganglion shows no significant findings. H&E ( e ) and IHC for HSV-1 ( f ). ( g , h ) Trigeminal ganglion of an unimmunized mouse: Meningitis is present in the meninges surrounding the trigeminal ganglion (arrow), and occasional neurons exhibit strong immunolabeling for HSV-1. H&E ( g ) and IHC for HSV1 ( h ). Scale bar for ( e – h ), 20 µm.

Article Snippet: To determine HSV-1 DNA copy number, we ran purified HSV-1 DNA (ATCC, VR-539DQ) or adipsin DNA (BioChain, Princeton, NJ, USA, D1334149) as standards.

Techniques: Infection, Immunolabeling