A lipidic delivery system of a triple vaccine adjuvant enhances mucosal immunity following nasal administration in mice

Ellen K. Wasan, Jaweria Syeda, Stacy Strom, Jacqueline Cawthray, Robert Hancock, Kishor M. Wasan, Volker Gerdts

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

We previously developed an highly efficacious combination adjuvant comprised of innate defense regulator (IDR)-1002 peptide, poly(I:C) and polyphosphazene (TriAdj). Here we aimed to design and test the in vivo efficacy of a mucoadhesive nasal formulation of this adjuvant. To determine the physical properties of the formulation, the effect of addition of each individual component was characterised by gel electrophoresis and fluorescence quenching using rhodamine-poly(I:C). Cationic liposomes comprised of didodecyl dimethylammonium bromide (DDAB), dioleoyl phosphatidylethanolamine (DOPE) (50:50 or 75:25 mol:mol) and DDAB, L-α-phosphatidylcholine (egg PC) and DOPE (40:50:10 mol:mol:mol) were prepared by the thin-film extrusion method. The liposomes and TriAdj were combined by simple mixing. The formed complex (L-TriAdj) was characterized by dynamic light scattering, zeta potential, and mucin interactions. We found that IDR-1002 peptide, polyphosphazene and poly(I:C) self-assembled in solution forming an anionic complex. Exposure of RAW267.4 mouse macrophage cells to TriAdj alone vs. L-TriAdj indicated that DDAB/DOPE (50:50) and DDAB/EPC/cholesterol (40:50:10) complexation reduced TriAdj toxicity. Next, TriAdj-containing cationic liposomes were prepared at several molar ratios to determine optimal size, stability and desired positive charge. Transmission electron microscopy showed rearrangement of lipid structures on binding of liposomes to TriAdj and to mucin. Stable particles (<200 nm over 24 h) showed mucin binding of DDAB/DOPE + TriAdj was greater than DDAB/EPC/DOPE + TriAdj. To verify in vivo efficacy, mice were administered the DDAB/DOPE + TriAdj complex intranasally with ovalbumin as the antigen, and the immunogenic response was measured by ELISA (serum IgG1, IgG2a, IgA) and ELISpot assays (splenocyte IL-5, IFN-γ). Mice administered adjuvant showed a significantly greater immune response with L-TriAdj than TriAdj alone, with a dose-response proportionate to the triple adjuvant content, and an overall balanced Th1/Th2 immune response representing both systemic and mucosal immunity.

LanguageEnglish
Pages1503-1515
Number of pages13
JournalVaccine
Volume37
Issue number11
DOIs
Publication statusPublished - 7 Mar 2019
Externally publishedYes

Keywords

  • Adjuvant
  • Intranasal delivery
  • Liposomes
  • Nanoparticles
  • Self-assembled systems
  • Vaccine

ASJC Scopus subject areas

  • Molecular Medicine
  • Immunology and Microbiology(all)
  • veterinary(all)
  • Public Health, Environmental and Occupational Health
  • Infectious Diseases

Cite this

Wasan, Ellen K. ; Syeda, Jaweria ; Strom, Stacy ; Cawthray, Jacqueline ; Hancock, Robert ; Wasan, Kishor M. ; Gerdts, Volker. / A lipidic delivery system of a triple vaccine adjuvant enhances mucosal immunity following nasal administration in mice. In: Vaccine. 2019 ; Vol. 37, No. 11. pp. 1503-1515.
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A lipidic delivery system of a triple vaccine adjuvant enhances mucosal immunity following nasal administration in mice. / Wasan, Ellen K.; Syeda, Jaweria; Strom, Stacy; Cawthray, Jacqueline; Hancock, Robert; Wasan, Kishor M.; Gerdts, Volker.

In: Vaccine, Vol. 37, No. 11, 07.03.2019, p. 1503-1515.

Research output: Contribution to journalArticle

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T1 - A lipidic delivery system of a triple vaccine adjuvant enhances mucosal immunity following nasal administration in mice

AU - Wasan, Ellen K.

AU - Syeda, Jaweria

AU - Strom, Stacy

AU - Cawthray, Jacqueline

AU - Hancock, Robert

AU - Wasan, Kishor M.

AU - Gerdts, Volker

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AB - We previously developed an highly efficacious combination adjuvant comprised of innate defense regulator (IDR)-1002 peptide, poly(I:C) and polyphosphazene (TriAdj). Here we aimed to design and test the in vivo efficacy of a mucoadhesive nasal formulation of this adjuvant. To determine the physical properties of the formulation, the effect of addition of each individual component was characterised by gel electrophoresis and fluorescence quenching using rhodamine-poly(I:C). Cationic liposomes comprised of didodecyl dimethylammonium bromide (DDAB), dioleoyl phosphatidylethanolamine (DOPE) (50:50 or 75:25 mol:mol) and DDAB, L-α-phosphatidylcholine (egg PC) and DOPE (40:50:10 mol:mol:mol) were prepared by the thin-film extrusion method. The liposomes and TriAdj were combined by simple mixing. The formed complex (L-TriAdj) was characterized by dynamic light scattering, zeta potential, and mucin interactions. We found that IDR-1002 peptide, polyphosphazene and poly(I:C) self-assembled in solution forming an anionic complex. Exposure of RAW267.4 mouse macrophage cells to TriAdj alone vs. L-TriAdj indicated that DDAB/DOPE (50:50) and DDAB/EPC/cholesterol (40:50:10) complexation reduced TriAdj toxicity. Next, TriAdj-containing cationic liposomes were prepared at several molar ratios to determine optimal size, stability and desired positive charge. Transmission electron microscopy showed rearrangement of lipid structures on binding of liposomes to TriAdj and to mucin. Stable particles (<200 nm over 24 h) showed mucin binding of DDAB/DOPE + TriAdj was greater than DDAB/EPC/DOPE + TriAdj. To verify in vivo efficacy, mice were administered the DDAB/DOPE + TriAdj complex intranasally with ovalbumin as the antigen, and the immunogenic response was measured by ELISA (serum IgG1, IgG2a, IgA) and ELISpot assays (splenocyte IL-5, IFN-γ). Mice administered adjuvant showed a significantly greater immune response with L-TriAdj than TriAdj alone, with a dose-response proportionate to the triple adjuvant content, and an overall balanced Th1/Th2 immune response representing both systemic and mucosal immunity.

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KW - Intranasal delivery

KW - Liposomes

KW - Nanoparticles

KW - Self-assembled systems

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