Abstract

The effectiveness of vaccines is enhanced by adding adjuvants. Furthermore, the selection of an inoculation route depends on the type of adjuvant used and is important for achieving optimum vaccine efficacy. We investigated the immunological differences between two types of vaccines—spike protein from the Middle East respiratory syndrome virus and inactivated influenza virus vaccine, in combination with a single-stranded RNA adjuvant—administered through various routes (intramuscular, intradermal, and intranasal) to BALB/c mice. Intramuscular immunization with the RNA adjuvant-formulated spike protein elicited the highest humoral immune response, characterized by IgG1 and neutralizing antibody production. Although intranasal immunization did not elicit a humoral response, it showed extensive T-cell activation through large-scale induction of interferon-γ- and interleukin-2-secreting cells, as well as CD4+ T-cell activation in mouse splenocytes. Moreover, only intranasal immunization induced IgA production. When immunized with the inactivated influenza vaccine, administration of the RNA adjuvant via all routes led to protection after viral challenge, regardless of the presence of a vaccine-specific antibody. Therefore, the inoculation route should depend on the type of immune response needed; i.e., the intramuscular route is suitable for eliciting a humoral immune response, whereas the intranasal route is useful for T-cell activation and IgA induction.

Highlights

  • Vaccines are currently one of the most effective and practical ways to protect the human body against various infectious diseases [1,2]

  • We formulated an RNA adjuvant with spike (S) protein-subunit vaccine for Middle East respiratory syndrome (MERS) coronavirus (CoV) and inactivated the influenza virus vaccine to investigate how the immune response differs according to the route of inoculation

  • The concentrations of interferon γ (IFN-γ), interleukin-2 (IL-2), IL-6, and tumor necrosis factor α (TNF-α) were detected with Enzyme-Linked Immunosorbent Assay (ELISA) kits (Invitrogen; Thermo Fisher Scientific Inc., Waltham, MA, USA), according to the manufacturer’s instructions. The concentrations of these cytokines were calculated according to standard curves, and the obtained results are shown as the amount of IFN-γ, IL-2, IL-6, and TNF-α per mL of supernatant

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Summary

Introduction

Vaccines are currently one of the most effective and practical ways to protect the human body against various infectious diseases [1,2]. Vaccinations prevent infectious disease because the pathogen-specific immune system, which has been previously trained, can eradicate pathogenic antigens when exposed to live pathogens [3,4]. Aluminum potassium sulfate, referred to as alum, induces a strong innate immune response through the influx of neutrophils, eosinophils, natural killer cells, CD11b+ monocytes, and dendritic cells at the injection site [11]. Single-stranded RNA (ssRNA) can act as an agonist of Toll-like receptor 7/8 (TLR-7/8) and retinoic acid-inducible gene I to induce an innate immune response [16]. We formulated an RNA adjuvant with spike (S) protein-subunit vaccine for Middle East respiratory syndrome (MERS) coronavirus (CoV) and inactivated the influenza virus vaccine to investigate how the immune response differs according to the route of inoculation. We assessed which administration routes are most effective for the RNA-adjuvant-formulated vaccination

Materials and Methods
Vaccines
In Vitro Transcription and RNA Purification
Immunization
Serum Collection
2.11. Flow Cytometry
2.12. Real-Time PCR for Virus Titration
Findings
World Health Organization
Full Text
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