Optimising the oil phases of aluminium hydrogel-stabilised emulsions for stable, safe and efficient vaccine adjuvant

doi:10.1007/s11705-021-2123-1

Front. Chem. Sci. Eng.

2022, Vol. 16

Issue (6) : 973-984 https://doi.org/10.1007/s11705-021-2123-1

RESEARCH ARTICLE

Optimising the oil phases of aluminium hydrogel-stabilised emulsions for stable, safe and efficient vaccine adjuvant

Lili Yuan¹, Xiao-Dong Gao¹(

), Yufei Xia^2,^3,⁴(

)

¹. Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China
². State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, China
³. University of Chinese Academy of Sciences, Beijing 100049, China
⁴. Innovation Academy for Green Manufacture, Chinese Academy of Sciences, Beijing 100190, China

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Abstract

To increase antibody secretion and dose sparing, squalene-in-water aluminium hydrogel (alum)-stabilised emulsions (ASEs) have been developed, which offer increased surface areas and cellular interactions for higher antigen loading and enhanced immune responses. Nevertheless, the squalene (oil) in previous attempts suffered from limited oxidation resistance, thus, safety and stability were compromised. From a clinical translational perspective, it is imperative to screen the optimal oils for enhanced emulsion adjuvants. Here, because of the varying oleic to linoleic acid ratio, soybean oil, peanut oil, and olive oil were utilised as oil phases in the preparation of aluminium hydrogel-stabilised squalene-in-water emulsions, which were then screened for their stability and immunogenicity. Additionally, the underlying mechanisms of oil phases and emulsion stability were unravelled, which showed that a higher oleic to linoleic acid ratio increased anti-oxidative capabilities but reduced the long-term storage stability owing to the relatively low zeta potential of the prepared droplets. As a result, compared with squalene-in-water ASEs, soybean-in-water ASEs exhibited comparable immune responses and enhanced stability. By optimising the oil phase of the emulsion adjuvants, this work may offer an alternative strategy for safe, stable, and effective emulsion adjuvants.

Keywords pickering emulsion vaccine adjuvant alum-stabilised emulsion oleic to linoleic acid ratio stability

Corresponding Author(s): Xiao-Dong Gao,Yufei Xia

Online First Date: 14 January 2022 Issue Date: 28 June 2022

Cite this article:

Lili Yuan,Xiao-Dong Gao,Yufei Xia. Optimising the oil phases of aluminium hydrogel-stabilised emulsions for stable, safe and efficient vaccine adjuvant[J]. Front. Chem. Sci. Eng., 2022, 16(6): 973-984.

URL:

https://academic.hep.com.cn/fcse/EN/10.1007/s11705-021-2123-1
https://academic.hep.com.cn/fcse/EN/Y2022/V16/I6/973

Tab.1 Selected oils for ASEs optimisations

Tab.2 Marking standard of stability evaluation scale

Fig.1 (a) Confocal images of ASEs and (b) ASE-loading antigen efficiency. OVA and the surface alum were labelled with Cy5 (red) and Lumogallion (green), respectively. Scale bar= 2 µm. Data were shown as mean ± s.e.m. (n = 3).

Tab.3 Optimal formulations and characterisations on the ASEs ^a)

Fig.2 Antigen reservoir effect: (a) in vivo images and (b) quantitative fluorescent intensity of Cy5-OVA persistence at the injection sites. Data were shown as mean ± s.e.m. (n = 6).

Fig.3 Serum OVA-specific IgG titer on Day 28. Data were shown as mean ± s.e.m. (n = 6) and analysed by one-way ANOVA.

Fig.4 Cytokine profile and memory T cell activations: (a) IFN-γ, (b) IL-4 levels in supernatant of ex vivo stimulated splenocytes, (c) effector memory T cells (CD44^high CD62L^low), and (d) central memory T cells (CD44^high CD62L^high) among CD3⁺ cells. Data were shown as mean ± s.e.m. (n = 6) and analysed by one-way ANOVA.

Fig.5 Biocompatibility evaluations via H&E staining of vital organ sections and injection sites (muscle) from BALB/c mice. Scale bar= 100 µm.

Fig.6 (a) IL-6, (b) IL-8, and (c) IL-10 levels in supernatant of ex vivo stimulated splenocytes. Data were shown as mean ± s.e.m. (n = 6) and analysed by one-way ANOVA.

Tab.4 Biochemical parameters in the serum

Fig.7 Storage stability of ASEs. (a) Optical micrographs of ASEs at the indicated temperatures on 30 days. Scale bar= 100 µm. For optical microscopy determination, the images were acquired with 40 × magnification. (b–d) Size of ASEs from Day 0 to Day 30 of storage at (b) 4 °C, (c) 25 °C, and (d) 37 °C. Data were shown as the mean ± s.e.m. (n = 3).

Tab.5 Oxidation induction period of the oils and ASEs

Tab.6 Likert chart and factor analysis on the stability of the ASEs (mean ± s.e.m.)

Fig.8 Quantification and characterisation of the stability of ASEs: (a) aggregation time of micrographs, (b) size change degree, and (c) zeta potential change degree. Data were shown as the mean ± s.e.m. (n = 9, ∗p<0.05, ∗∗p<0.01, ∗∗∗p<0.001) and analysed by one-way ANOVA.

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