Adenovirus type 5 SARS-CoV-2 vaccines delivered orally or intranasally reduced disease severity and transmission in a hamster model

Stephanie N Langel, Susan Johnson, Clarissa I Martinez, Sarah N Tedjakusuma, Nadine Peinovich, Emery G Dora, Philip J Kuehl, Hammad Irshad, Edward G Barrett, Adam D Werts, Sean N Tucker, Stephanie N Langel, Susan Johnson, Clarissa I Martinez, Sarah N Tedjakusuma, Nadine Peinovich, Emery G Dora, Philip J Kuehl, Hammad Irshad, Edward G Barrett, Adam D Werts, Sean N Tucker

Abstract

Transmission-blocking strategies that slow the spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and protect against coronavirus disease 2019 (COVID-19) are needed. We have developed an orally delivered adenovirus type 5-vectored SARS-CoV-2 vaccine candidate that expresses the spike protein. Here, we demonstrated that hamsters vaccinated by the oral or intranasal route had robust and cross-reactive antibody responses. We then induced a postvaccination infection by inoculating vaccinated hamsters with SARS-CoV-2. Orally or intranasally vaccinated hamsters had decreased viral RNA and infectious virus in the nose and lungs and experienced less lung pathology compared to mock-vaccinated hamsters after SARS-CoV-2 challenge. Naïve hamsters exposed in a unidirectional air flow chamber to mucosally vaccinated, SARS-CoV-2-infected hamsters also had lower nasal swab viral RNA and exhibited fewer clinical symptoms than control animals, suggesting that the mucosal route reduced viral transmission. The same platform encoding the SARS-CoV-2 spike and nucleocapsid proteins elicited mucosal cross-reactive SARS-CoV-2-specific IgA responses in a phase 1 clinical trial (NCT04563702). Our data demonstrate that mucosal immunization is a viable strategy to decrease SARS-CoV-2 disease and airborne transmission.

Figures

Fig. 1.. Oral and intranasal r-Ad-S vaccination…
Fig. 1.. Oral and intranasal r-Ad-S vaccination induced robust IgG and IgA antibodies in golden hamsters.
Index hamsters were immunized with oral r-Ad-S, intranasal (IN) r-Ad-S, intramuscular (IM) spike protein (S) or mock (oral) (n = 4 per group) and inoculated with SARS-CoV-2 seven weeks later. (A) Experimental design schematic. Figure created with BioRender.com (B) Serum anti-spike protein IgG antibody endpoint titers were measured at weeks 0, 3 and 6 post immunization by enzyme-linked immunosorbent assay (ELISA). (C) Serum anti-spike protein IgG antibody endpoint titers were measured at week 6 post immunization against the spike protein of the Wuhan, Beta, and Delta SARS-CoV-2 variants by ELISA. (D) Surrogate neutralizing antibodies (antibodies capable of blocking binding of SARS-CoV-2 spike protein to ACE2) titers were measured in serum at week 6. (E) Serum anti-spike protein IgA antibody titers were measured (MSD arbitrary units (AU)/sample) at weeks 0, 3, and 6 post immunization using the MSD platform and normalized to day 0 AU values. (F) anti-spike protein IgA antibodies were measured in BAL fluid using the MSD platform. Data were analyzed by a one-way ANOVA and Dunnett’s multiple comparisons. Comparisons were made between vaccinated groups and mock (oral) controls for (B, D, E and F). Error bars represent the standard error of the mean (SEM). *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.
Fig. 2.. Oral and intranasal r-Ad-S vaccination…
Fig. 2.. Oral and intranasal r-Ad-S vaccination decreased SARS-CoV-2 RNA and infectious virus in nasal swabs and lungs.
(A) Nasal swabs were collected on days 1, 3, and 5 post-inoculation in index hamsters (n = 4 per group) and viral genomic RNA loads in these samples were determined by quantitative reverse transcription PCR (qRT-PCR) of the N gene. (B) Nasal swabs were collected on day 1 post inoculation and infectious virus titers were determined by TCID50. (C) Lung tissue was collected at necropsy (day 5) and RNA was isolated for SARS-CoV-2 detection by qRT-PCR of the N gene and (D) infectious viral titers by TCID50. The dotted line represents Lower Limit of Quantification (LLOQ), with data below the limit of detection plotted at ½ LLOQ. Data were analyzed by a one-way ANOVA and Dunnett’s multiple comparisons (A to C) or Kruskal-Wallis with Dunn’s multiple comparisons (D). Comparisons were made between vaccinated groups and mock (oral) controls. Error bars represent the SEM. *P<0.05, **P<0.01.
Fig. 3.. Oral and intranasal r-Ad-S vaccination…
Fig. 3.. Oral and intranasal r-Ad-S vaccination reduced disease indicators, including body weight loss, lung weight, and lung pathology scores in index hamsters.
(A) Daily weight change and (B) terminal body weights were determined by the percent of day 0 weight (relative to SARS-CoV-2 inoculation) for index hamsters (n = 4 per group). (C) relative terminal lung weights and (D) lung pathology scores were determined. Severity grade for red discoloration of the lung was based on a 0 to 4 scale indicating percent of whole lung affected: none (no grade), minimal (1), mild (2), moderate (3), marked (4) correlating to 0, 1 to 25, 26 to 50, 51 to 75, or 76 to 100% affected, respectively. Data were analyzed by a one-way ANOVA and Dunnett’s multiple comparisons. Comparisons were made between vaccinated groups and mock (oral) controls. Error bars represent the SEM. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.
Fig. 4.. Oral and intranasal SARS-CoV-2 vaccination…
Fig. 4.. Oral and intranasal SARS-CoV-2 vaccination decreased SARS-CoV-2 aerosol transmission.
(A) Nasal swabs were collected on days 1 and 3 post-inoculation from index animals (n = 4 per group) and from naïve animals (n = 16 per group) on days 1, 3, and (B) 5 after exposure to index, infected hamsters in airborne transmission chambers. Viral genomic RNA loads in these samples were determined by qRT-PCR of the N gene. (C) Lung tissue was collected at necropsy (day 5) and RNA was isolated for SARS-CoV-2 detection by qRT-PCR of the N gene. (D) Infectious viral titers in lung tissue were determined by TCID50. In (A to D) The dotted line represents the LLOQ, with data below the limit of detection plotted at ½ LLOQ. (E) Terminal body weights were determined by the percent weight change versus day 0 (relative to SARS-CoV-2 inoculation). (F) Terminal lung weights and (G) lung pathology scores were determined. Severity grade for red discoloration of the lung was based on a 0 to 4 scale as in Fig. 3D. In (E to G), data were analyzed by a one-way ANOVA and Dunnett’s multiple comparisons. Comparisons were made between vaccinated groups and mock (oral) controls. For all panels, error bars represent the SEM. *P <0.05, **P <0.01, ***P < 0.001, ****P<0.0001.
Fig. 5.. Cross reactive anti-spike protein IgA…
Fig. 5.. Cross reactive anti-spike protein IgA in the mucosal and salivary compartments is elicited by oral Ad5 vaccination in a subset of participants.
(A and B) Fold change (day 29 versus day 1) in spike (S)-, nucleocapsid (N)-, or receptor binding domain (RBD)-specific IgA antibodies in nasal (A) and saliva (B) samples were measured by the MSD platform (n = 35). (C and D). The fold change in IgA specific to endemic human coronaviruses was measured in both nasal (C) and saliva (D) samples respectively, in the responders (n = 12 or n=7 respectively) from (A) and (B). The red dotted line represents a two-fold change, by which individuals were classified as responders.
https://www.ncbi.nlm.nih.gov/pmc/articles/instance/9097881/bin/keyimage.jpg

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