Immune Netw.  2012 Dec;12(6):261-268. 10.4110/in.2012.12.6.261.

Immunogenicity and Protective Efficacy of a Dual Subunit Vaccine Against Respiratory Syncytial Virus and Influenza Virus

• Park MH
• Chang J

• KMID: 2150758
• DOI: http://doi.org/10.4110/in.2012.12.6.261

Abstract

Respiratory syncytial virus (RSV) and influenza virus are the most significant pathogens causing respiratory tract diseases. Composite vaccines are useful in reducing the number of vaccination and confer protection against multiple infectious agents. In this study, we generated fusion of RSV G protein core fragment (amino acid residues 131 to 230) and influenza HA1 globular head domain (amino acid residues 62 to 284) as a dual vaccine candidate. This fusion protein, Gcf-HA1, was bacterially expressed, purified by metal resin affinity chromatography, and refolded in PBS. BALB/c mice were intranasally immunized with Gcf-HA1 in combination with a mucosal adjuvant, cholera toxin (CT). Both serum IgG and mucosal IgA responses specific to Gcf and HA1 were significantly increased in Gcf-HA1/CT-vaccinated mice. To determine the protective efficacy of Gcf-HA1/CT vaccine, immunized mice were challenged with RSV (A2 strain) or influenza virus (A/PR/8/34). Neither detectable viral replication nor pathology was observed in the lungs of the immune mice. These results demonstrate that immunity induced by intranasal Gcf-HA1/CT immunization confers complete protection against both RSV and homologous influenza virus infection, suggesting our Gcf-HA1 vaccine candidate could be further developed as a dual subunit vaccine against RSV and influenza virus.

Keyword

Respiratory syncytial virus; Influenza virus; G glycoprotein; Hemagglutinin; Dual vaccine; Intranasal immunization; Protective immunity

MeSH Terms

Animals
Cholera Toxin
Chorionic Gonadotropin, beta Subunit, Human
Chromatography, Affinity
GTP-Binding Proteins
Head
Hemagglutinins
Immunization
Immunoglobulin A
Immunoglobulin G
Influenza, Human
Lung
Mice
Orthomyxoviridae
Peptide Fragments
Respiratory Syncytial Viruses
Respiratory Tract Diseases
Vaccination
Vaccines
Cholera Toxin
Chorionic Gonadotropin, beta Subunit, Human
GTP-Binding Proteins
Hemagglutinins
Immunoglobulin A
Immunoglobulin G
Peptide Fragments
Vaccines

Figure

• 

  Figure 1 Preparation of Gcf-HA1 subunit vaccine. Expression of RSV G protein core fragment (Gcf: amino acid 131 to 230) fused with influenza HA1 domain (amino acid 76 to 284) was performed in pET-21d-Gcf-HA1-transformed E. coil BL21(DE3). Expressed fusion protein was purified from inclusion body by immobilized metal ion affinity chromatography. Then, Gcf-HA1 fusion protein was refolded in PBS. (A) Sequence structure of pET-21d-Gcf-HA1 expression vector was shown in the diagram. (B) The expression of Gcf-HA1 fusion protein was confirmed by SDS-PAGE of samples from purification steps.

• 

  Figure 2 Serum antibody responses induced by Gcf-HA1/CT immunization. BALB/c mice were immunized twice on day 0 and 14 by intranasal (i.n) injection of Gcf-HA1 fusion protein in combination with CT. (A) Vaccination and challenge experiment scheme. (B) Serum IgG antibody responses specific for Gcf were measured by ELISA 3 weeks after boosting immunization. (C) Serum IgG antibody titers specific for HA were measured by ELISA 3 weeks after boosting immunization. N.D., not detected.

• 

  Figure 3 Mucosal antibody responses induced by Gcf-HA1/CT immunization. (A) Mucosal IgA antibody responses specific for Gcf in BAL fluid were measured by ELISA 5 days after RSV challenge. (B) Mucosal IgA antibody titers specific for HA were measured 5 days after influenza PR8 challenge. N.D., not detected.

• 

  Figure 4 Protective efficacy of Gcf-HA1/CT vaccination against RSV challenge. The immune mice were challenged intranasally with 106 PFU of RSV A2. (A) Lung homogenates were prepared and lung viral titers were measured by standard plaque assay with HEp-2 cells. (B) The numbers of neutrophils and eosinophils in BAL fluids were measured by flow cytometry at day 5 following RSV challenge. N.D., not detected.

• 

  Figure 5 Protective efficacy of Gcf-HA1/CT vaccination against influenza virus infection. The immune mice were challenged with 10 LD50 of influenza PR8 virus. (A) Lung homogenates were prepared and lung viral titers were measured by standard plaque assay with MDCK cells. (B) Mice were monitored for weight loss each day after challenge.

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