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Expression and Purification of Virus-like Particles for Vaccination
Published on: June 2, 2016
Vaccination with a human parainfluenza virus type 3 chimeric FHN glycoprotein formulated with a combination adjuvant
R Garg1, R Brownlie1, L Latimer1
1VIDO-Intervac, University of Saskatchewan, Saskatoon, SK S7N 5E3, Canada.
Insights
A novel chimeric FHN glycoprotein vaccine candidate shows promise for preventing human parainfluenza virus type 3 (PIV3) lower respiratory infections in infants. Immunization induced strong neutralizing antibodies and protection against PIV3 challenge.
Area of Science:
- Virology
- Vaccinology
- Immunology
Background:
- Human parainfluenza virus type 3 (PIV3) is a leading cause of severe lower respiratory tract infections in infants and young children.
- Currently, no licensed vaccine is available to prevent PIV3 infections.
Purpose of the Study:
- To develop an effective subunit vaccine against PIV3.
- To evaluate the immunogenicity and protective efficacy of a chimeric FHN glycoprotein vaccine candidate.
Main Methods:
- Expression and purification of a chimeric FHN glycoprotein and individual F and HN proteins in mammalian cells.
- Immunization of mice, cotton rats, and hamsters with FHN or F+HN proteins formulated with TriAdj adjuvant via intramuscular or intranasal routes.
- Assessment of systemic and mucosal immune responses, including virus-neutralizing antibodies and IgA.
- Evaluation of protection against PIV3 challenge by measuring viral load.
Main Results:
- The FHN/TriAdj formulation elicited significantly higher levels of systemic virus-neutralizing antibodies compared to the F+HN/TriAdj formulation.
- Intranasal administration of FHN/TriAdj induced mucosal IgA production and serum neutralizing antibodies.
- All animals immunized with FHN/TriAdj were protected against PIV3 challenge, with no detectable virus post-exposure.
- FHN/TriAdj induced protective immunity against PIV3 in multiple animal models.
Conclusions:
- The chimeric FHN protein formulated with TriAdj demonstrates potential as a safe and effective PIV3 vaccine.
- Both systemic and mucosal immunization routes with FHN/TriAdj confer protection against PIV3 infection.
Abstract:
Human parainfluenza virus type 3 (PIV3) is a major cause of lower respiratory disease i.e. bronchitis, bronchiolitis or pneumonia, in infants and young children. Presently there is no licensed vaccine against PIV3. To produce an effective subunit vaccine, a chimeric FHN glycoprotein consisting of the N-terminal ectodomain of the fusion (F) protein linked to the haemagglutinin-neuraminidase (HN) protein without transmembrane domain, and secreted forms of the individual F and HN glycoproteins, were expressed in mammalian cells and purified. Mice and cotton rats were immunized intramuscularly (IM) with FHN or both F and HN proteins (F + HN), formulated with poly(I:C) and an innate defense regulator peptide in polyphosphazene (TriAdj). Significantly higher levels of systemic virus-neutralizing antibodies were observed in mice and cotton rats immunized with FHN/TriAdj when compared to animals immunized with the combination of F and HN proteins (F + HN/TriAdj). As PIV3 is a pneumotropic virus, another goal is to produce an effective mucosal subunit vaccine. Intranasal (IN) administration with FHN/TriAdj resulted in mucosal IgA production in the lung and virus neutralizing antibodies in the sera. After PIV3 challenge no virus was detected in cotton rats immunized with FHN/TriAdj regardless of the route of delivery. Protective immunity against PIV3 was also induced by FHN/TriAdj in hamsters. In conclusion, the FHN protein formulated with TriAdj has potential for development of a safe and effective vaccine against PIV3.
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