Preparation of Oral Vaccine Carriers by Combining Morphology Control and Plating Modification

doi:10.13523/j.cb.2204006

China Biotechnology

2022, Vol. 42

Issue (8): 40-51 DOI: 10.13523/j.cb.2204006

Preparation of Oral Vaccine Carriers by Combining Morphology Control and Plating Modification

HU Wei^1,²,WU Jie²,HIDEKI Nakanishi^1,^**(

)

1. Key Laboratory of Carbohydrate Chemistry and Biotechnology of 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

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Abstract

Oral vaccines have attracted much attention due to their advantages of high patient compliance, reduced generation of harmful waste, convenient vaccination, and their ability to cause mucosal immunity. However, unfavorable conditions such as the acidic environment of the stomach, proteases, intestinal mucus and tight junctions between intestinal epithelial cells make the gap between oral vaccines and injectable vaccines too large, which restricts the immune effect of oral vaccines. In order to improve the immune effect of oral vaccines, we combined morphology control and coating modification strategies to prepare a new type of oral vaccine carrier. Specifically, polylactic acid-glycolic acid copolymer (PLGA) rod-shaped particles were prepared by combining emulsion solvent evaporation method with fast membrane emulsification method, then β-glucan that enhances immune response and thiolated hydroxypropyl methyl cellulose phthalate (T-HPMCP) with higher degradation pH and stronger adhesion with intestinal epithelium were used for coating modification of PLGA rod-shaped particles. In the preparation of PLGA rod-shaped particles, the effects of PBS concentration and polyvinyl alcohol (PVA) concentration in the outer aqueous phase on the preparation of PLGA rod-shaped particles were explored. It is found that the deformation degree of PLGA particles first increased and then decreased with the increase of PBS concentration, but the deformation degree of PLGA particles always increased with the increase of PVA concentration. This is because PBS forms an electric double layer with -COO^- of PLGA, which makes the emulsion more stable and makes deformation more difficult to occur. Finally, the optimal formula was determined to prepare PLGA rod-shaped particles with a length of 2-4 μm and a width of 1-2 μm suitable for the uptake of small intestinal epithelial cells. It is found that the protein entrapment rate of PLGA rod-shaped particles is higher than that of PLGA spherical particles because the electric double layer makes the emulsion more stable. The results of in vitro experiments show that the vaccine carrier modified by T-HPMCP is stable in an acidic environment and the amount of released protein is negligible, which can prevent the antigen from being corroded by the acidic environment and is conducive to the protection of antigen activity. It can be decomposed at pH ≥ 7.4 and then release antigen. The results of cell experiments show that its special rod-shaped morphology can be taken up by Caco-2 cells faster and can be rapidly transported by the Caco-2 cell monolayer model constructed in transwell chambers, and the modification of β-glucan can also promote dendritic cells (DCs) secretion of surface molecules MHC-I, MHC-II and CD80 to activate DCs. The results of animal experiments showed that PLGA rod-shaped particles modified by β-glucan and T-HPCMP could increase the levels of OVA-specific IgA and IgG antibodies, which reached their maximum on the 28th day, and they promoted immune central memory T cells and CD8⁺ effects generation of memory T cells. In conclusion, the prepared coated PLGA rod-shaped particles can improve the immune response of the body as an oral vaccine carrier, thereby producing a better immune effect and providing new materials and ideas for the research of oral vaccines.

Key words： Oral vaccine PLGA rod-shaped particles β-glucan Dendritic cells (DCs) Mucosal immunity

Received: 04 April 2022 Published: 07 September 2022

ZTFLH:

Q819

Corresponding Authors: Nakanishi HIDEKI E-mail: hideki@jiangnan.edu.cn

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Cite this article:

HU Wei,WU Jie,HIDEKI Nakanishi. Preparation of Oral Vaccine Carriers by Combining Morphology Control and Plating Modification. China Biotechnology, 2022, 42(8): 40-51.

URL:

https://manu60.magtech.com.cn/biotech/10.13523/j.cb.2204006 OR https://manu60.magtech.com.cn/biotech/Y2022/V42/I8/40

Fig.1 Schematic diagram of the preparation flow chart of coated PLGA rod-shaped particl

Table 1 Each immunization dose per mouse

Fig.2 PLGA particles prepared under different PBS concentrations (a) 3 mmol (b) 5 mmol (c) 10 mmol (d) 13 mmol (e) 15 mmol (f) Relationship between PBS concentration and PLGA particle aspect ratio

Fig.3 PLGA particles prepared under different PVA concentrations (a) 0.3% (b) 0.5% (c) 0.75% (d) 1% (e) 1.5% (f) 2% (g) Relationship between PVA concentration and PLGA particle aspect ratio

Fig.4 XRD patterns of PLGA rod-shaped particles(a), PLGA spherical particles(b) and PLGA particles with different morphologies(c)

Fig.5 BET specific surface area(a) and protein entrapment rates(b) of PLGA particles with different morphologies

Fig.6 IR spectrum(a)and NMR image (b) of T-HPMCP

Fig.7 Plating potential distribution diagram

Fig.8 In vitro protein release profiles of PLGA particles with and without pH-sensitive shells

Fig.9 Investigating the permeation rate of PLGA particles through the Caco-2 cell monolayer model

Fig.10 Confocal characterization of Caco-2 cells uptake of coated PLGA rod-shaped particles(a)and flow cytometry characterizes the uptake of PLGA particles with different morphologies by Caco-2 cells(b)

Fig.11 Activation effect of PLGA particles with different surface coatings on the surface molecules of DC (a) CD11c⁺MHCI⁺ (b) CD11c⁺MHCII⁺ (c) CD11c⁺CD80⁺

Fig.12 OVA-specific antibody secretion (a)IgA (b)IgG

Fig.13 Memory T lymphocytes activation (a) CD4⁺CD44⁺CD62L⁺ (b) CD4⁺CD44⁺CD62L^- (c) CD8⁺CD44⁺CD62L⁺ (d) CD8⁺CD44⁺CD62L^-


[1]	Shamsuzzaman S, Ahmed T, Mannoor K, et al. Robust gut associated vaccine-specific antibody-secreting cell responses are detected at the mucosal surface of Bangladeshi subjects after immunization with an oral killed bivalent V. cholerae O1/O 139 whole cell cholera vaccine: comparison with other mucosal and systemic responses. Vaccine, 2009, 27(9): 1386-1392. doi: 10.1016/j.vaccine.2008.12.041 pmid: 19146897

[2]	Drucker D J. Advances in oral peptide therapeutics. Nature Reviews Drug Discovery, 2020, 19(4): 277-289. doi: 10.1038/s41573-019-0053-0 pmid: 31848464

[3]	Ferber S, Gonzalez R J, Cryer A M, et al. Immunology-guided biomaterial design for mucosal cancer vaccines. Advanced Materials, 2020, 32(13): 1903847. doi: 10.1002/adma.201903847

[4]	Lee K Y, Mooney D J. Alginate: properties and biomedical applications. Progress in Polymer Science, 2012, 37(1): 106-126. doi: 10.1016/j.progpolymsci.2011.06.003

[5]	Singh B, Maharjan S, Jiang T, et al. Attuning hydroxypropyl methylcellulose phthalate to oral delivery vehicle for effective and selective delivery of protein vaccine in ileum. Biomaterials, 2015, 59: 144-159. doi: 10.1016/j.biomaterials.2015.04.017

[6]	Banerjee A, Qi J P, Gogoi R, et al. Role of nanoparticle size, shape and surface chemistry in oral drug delivery. Journal of Controlled Release, 2016, 238: 176-185. doi: S0168-3659(16)30497-7 pmid: 27480450

[7]	Dendukuri D, Pregibon D C, Collins J, et al. Continuous-flow lithography for high-throughput microparticle synthesis. Nature Materials, 2006, 5(5): 365-369. doi: 10.1038/nmat1617

[8]	Dendukuri D, Tsoi K, Hatton T A, et al. Controlled synthesis of nonspherical microparticles using microfluidics. Langmuir: the ACS Journal of Surfaces and Colloids, 2005, 21(6): 2113-2116. doi: 10.1021/la047368k

[9]	Fan Q Z, Qi F, Miao C Y, et al. Direct and controllable preparation of uniform PLGA particles with various shapes and surface morphologies. Colloids and Surfaces A: Physicochemical and Engineering Aspects, 2016, 500: 177-185. doi: 10.1016/j.colsurfa.2016.04.028

[10]	de Smet R, Demoor T, Verschuere S, et al. Β-Glucan microparticles are good candidates for mucosal antigen delivery in oral vaccination. Journal of Controlled Release: Official Journal of the Controlled Release Society, 2013, 172(3): 671-678. doi: 10.1016/j.jconrel.2013.09.007

[11]	Liu H, Jia Z H, Yang C M, et al. Aluminum hydroxide colloid vaccine encapsulated in yeast shells with enhanced humoral and cellular immune responses. Biomaterials, 2018, 167: 32-43. doi: 10.1016/j.biomaterials.2018.03.014

[12]	Nan F F, Wu J, Qi F, et al. Preparation of uniform-sized colloidosomes based on chitosan-coated alginate particles and its application for oral insulin delivery. Journal of Materials Chemistry B, 2014, 2(42): 7403-7409. doi: 10.1039/C4TB01259C

[13]	Quan J S, Jiang H L, Kim E M, et al. pH-sensitive and mucoadhesive thiolated eudragit-coated chitosan microspheres. International Journal of Pharmaceutics, 2008, 359(1-2): 205-210. doi: 10.1016/j.ijpharm.2008.04.003 pmid: 18490120

[14]	Lv P P, Wei W, Yue H, et al. Porous quaternized chitosan nanoparticles containing paclitaxel nanocrystals improved therapeutic efficacy in non-small-cell lung cancer after oral administration. Biomacromolecules, 2011, 12(12): 4230-4239. doi: 10.1021/bm2010774

[15]	Heslinga M J, Mastria E M, Eniola-Adefeso O. Fabrication of biodegradable spheroidal microparticles for drug delivery applications. Journal of Controlled Release, 2009, 138(3): 235-242. doi: 10.1016/j.jconrel.2009.05.020 pmid: 19467275

[16]	Delon L, Gibson R J, Prestidge C A, et al. Mechanisms of uptake and transport of particulate formulations in the small intestine. Journal of Controlled Release, 2022, 343: 584-599. doi: 10.1016/j.jconrel.2022.02.006

[17]	Mohan S, Jayaprakash A, Jose S P. Fourier transform Raman and infrared spectral investigations of 2, 6-dimethyl-2, 5-heptadien-4-one. AIP Conference Proceedings, 2010, 1267(1): 1209-1210.

[18]	Xiong X B, Mahmud A, Uludağ H, et al. Conjugation of arginine-glycine-aspartic acid peptides to poly (ethylene oxide)-b-poly(ε-caprolactone) micelles for enhanced intracellular drug delivery to metastatic tumor cells. Biomacromolecules, 2007, 8(3): 874-884. doi: 10.1021/bm060967g

[19]	Sharma G, Valenta D T, Altman Y, et al. Polymer particle shape independently influences binding and internalization by macrophages. Journal of Controlled Release, 2010, 147(3): 408-412. doi: 10.1016/j.jconrel.2010.07.116

[20]	Huang H B, Ostroff G R, Lee C K, et al. Robust stimulation of humoral and cellular immune responses following vaccination with antigen-loaded beta-glucan particles. mBio, 2010, 1(3): e00164-e00110.

[21]	Cornel A M, Mimpen I L, Nierkens S. MHC class I downregulation in cancer: underlying mechanisms and potential targets for cancer immunotherapy. Cancers, 2020, 12(7): 1760. doi: 10.3390/cancers12071760

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