Radial porous SiO<sub>2</sub> nanoflowers potentiate the effect of antigen/adjuvant in antitumor immunotherapy

doi:10.1007/s11705-020-2034-6

Front. Chem. Sci. Eng.

2021, Vol. 15

Issue (5) : 1296-1311 https://doi.org/10.1007/s11705-020-2034-6

RESEARCH ARTICLE

Radial porous SiO₂ nanoflowers potentiate the effect of antigen/adjuvant in antitumor immunotherapy

Chuangnian Zhang¹, Ying Dong², Jing Gao², Xiaoli Wang¹(

), Yanjun Jiang²(

)

¹. Tianjin Key Laboratory of Biomaterial Research, Institute of Biomedical Engineering, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin 300192, China
². School of Chemical Engineering and Technology, Hebei University of Technology, Tianjin 300130, China

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Abstract

Here, we reported a cancer nanovaccine based on SiO₂ nanoflowers with a special radial pore structure, which greatly enhanced cross-presentation and induced the production of cytotoxic T lymphocyte cells secreting granzymes B and interferon-γ. The antigen ovalbumin was covalently conjugated onto the as-synthesized hierarchical SiO₂ nanoflowers, and the adjuvant cytosine-phosphate-guanine was electrostatically adsorbed into their radial pore by simple mixing before use. The nanovaccine exhibited excellent storage stability without antigen release after 27 days of incubation, negligible cytotoxicity to dendritic cells, and a high antigen loading capacity of 430 ± 66 mg·g⁻¹ support. Besides, the nanovaccine could be internalized by dendritic cells via multiple pathways. And the enhancement of antigen/adjuvant uptake and lysosome escape of antigen were observed. Noteworthy, in vitro culture of bone marrow-derived dendritic cells in the presence of nanovaccine proved the activation of dendritic cells and antigen cross-presentation as well as secretion of proinflammatory cytokines. Besides, in vivo study verified the targeting of nanovaccine to draining lymph nodes, the complete suppression of tumor in six out of ten mice, and the triggering of notable tumor growth delay. Overall, the present results indicated that the nanovaccine can be served as a potential therapeutic vaccine to treat cancer.

Keywords silica nanoflower antigen delivery cancer immunotherapy nanovaccine

Corresponding Author(s): Xiaoli Wang,Yanjun Jiang

Just Accepted Date: 03 March 2021 Online First Date: 12 April 2021 Issue Date: 30 August 2021

Cite this article:

Chuangnian Zhang,Ying Dong,Jing Gao, et al. Radial porous SiO₂ nanoflowers potentiate the effect of antigen/adjuvant in antitumor immunotherapy[J]. Front. Chem. Sci. Eng., 2021, 15(5): 1296-1311.

URL:

https://academic.hep.com.cn/fcse/EN/10.1007/s11705-020-2034-6
https://academic.hep.com.cn/fcse/EN/Y2021/V15/I5/1296

Fig.1 (a) Schematic diagram for the synthetic process of the nanovaccine (OVA@fSiO₂ + CpG); (b) SEM image; (c) TEM image; (d) the nitrogen adsorption-desorption isotherm; (e) the pore diameter distribution curve of fSiO₂-NH₂; (f) the effect of glutaraldehyde concentration on the OVA loading capacity in OVA@fSiO₂; (g) the surface zeta potential of fSiO₂ prior and after amination, activation and covalently conjugated with OVA; (h) dynamic light scattering size distribution of fSiO₂-NH₂ and OVA@fSiO₂; (i) Fourier transform infrared spectrum of fSiO₂-OH, fSiO₂-NH₂, fSiO₂-CHO and OVA@fSiO₂; (j–l) the stabilities of OVA@fSiO₂ in different pH solution.

Fig.2 (a) The cell viability of DC2.4 and RAW264.7 after incubating these cells for 48 h with OVA@fSiO₂, fresh culture medium was served as a negative control; (b) the internalization pathways of nanovaccine by DCs and the corresponding inhibitors; the uptake of (c, e) OVA and (d, f) CpG estimated by the MFI of DC2.4 cells after incubating the cells with OVA@fSiO₂ + CpG in the presence of various inhibitors, and their representative FACS histograms (Data were expressed as the mean±SD of five individual experiments, and the differences between groups were determined by one-way ANOVA followed by Tukey’s multiple comparison test, *P<0.05, **P<0.01, ***P<0.001).

Fig.3 (a) Images observed by CLSM after incubating BMDCs with OVA+ CpG or OVA@fSiO₂ + CpG for 24 h, the endo/lysosomes membrane was marked with Lamp-1-APC; (b) the colocalization analysis was conducted by Zeiss Zen 2008 Light Edition.

Fig.4 The expression of costimulatory markers: (a) CD40; (b) CD80; (c) CD86; (d) MHC II; (e) MHC I; (f) SIINFEKL-MHC I on pulsed BMDCs; (g) the cross-presentation of antigen OVA was appraised by the activity of b-galactosidase produced by B3Z cells using a colorimetric LacZ assay (OD405 nm); (h) the expression of CCR7 on pulsed BMDCs (Data were represented the mean±SD of three individual experiments, the differences between groups were determined by one-way ANOVA followed by Tukey’s multiple comparison test, ** P<0.01, *** P<0.001).

Fig.5 The released cytokine levels (IL-12, IL-1b, IL-6, TNF-a, IL-4 and IL-10) from BMDCs pulsed with PBS, OVA+ CpG or OVA@fSiO₂ + CpG (Data were represented the mean±SD of three individual experiments, the differences between groups were determined by one-way ANOVA followed by Tukey’s multiple comparison test, *** P<0.001).

Fig.6 The fluorescent signal of Cy7 at the administration site and left lymph nodes of mice vaccinated by OVA-Cy7 or OVA-Cy7@fSiO₂, lymph nodes indicated by red arrows.

Fig.7 (a) The images of fresh tumor tissue excised from each treated group at day 25; (b–d) the tumor growth curves of each mouse in each experimental group; (e) the average tumor volumes at the 25th day; (f) the average tumor weight of mice at the 25th day; (g) the body weight of mice over time in each experimental group (Data were shown as the mean±s. e. m of ten individual experiments and analyzed by unpaired student’s t-test, *P<0.05).

Fig.8 The percentages of (a) Ki67⁺, (b) CD44⁺CD62⁺ and (c) CD44⁺CD62^– among CD8⁺ T cells in mouse splenocyte; (d) the percentages of CD3⁺CD8⁺Granzyme B⁺ CTLs in mouse splenocyte; (e) IFN-g level of splenocyte supernatant after three days of antigen restimulation; (f) the serum IFN-g level tested by ELISA assay; (g–i) IL-2, IL-4 and IL-10 levels of splenocyte supernatant after three days of antigen restimulation; OVA-specific total (j) IgG titer, (k) IgG1 titer, and (l) IgG2a titer on day 25 were measured by ELISA assay (The data were shown as the means±s. e. m (n = 10) and analyzed using one-way ANOVA followed by Newman-Keuls multiple comparison test. *P<0.05, **P<0.01, ***P<0.001).

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