Eudragit L100修饰的铝锰双金属有机框架作为猪流行性腹泻灭活疫苗口服递送载体的初步评价

doi:10.11843/j.issn.0366-6964.2025.05.027

Abstract

Abstract:

This study aims to evaluate aluminum-manganese bimetallic organic framework nanoparticles modified with Eudragit L100 as an oral delivery vehicle for inactivated porcine epidemic diarrhea virus (PEDV). By utilizing the coordination interaction between metal ions (Al³⁺, Mn²⁺) and the organic ligand 2-aminoterephthalic acid, inactivated PEDV is encapsulated in situ within Al/Mn-MOF to form Al/Mn-MOF@PEDV nanoparticles. An anionic polymer, Eudragit L100, is then electrostatically attached to the surface of Al/Mn-MOF@PEDV nanoparticles, producing L100@Al/Mn-MOF@PEDV nanoparticles. The morphology is examined using scanning electron microscopy, the surface charge is measured by zeta potential meter, the composition is analyzed by fourier transform infrared spectroscopy, and the crystal structure is determined by X-ray Diffraction. The loading efficiency of inactivated PEDV is characterized using ELISA and antigen detection cards. The stability of L100@Al/Mn-MOF@PEDV nanoparticles in simulated gastric fluid and the release rate of inactivated PEDV are examined using ELISA. Macrophage uptake of the nanoparticles is observed using confocal laser scanning microscopy. The cytotoxicity of nanoparticles with different concentrations is determined by CCK8 method, and the effect on the secretion of immunomodulatory factors by macrophages is evaluated using ELISA kits. Results indicate successful preparation of L100@Al/Mn-MOF@PEDV nanoparticles, with sizes under 1 μm. At a PEDV/metal ion/organic ligand ratio of 0.42 ∶1 ∶1, the loading rate of inactivated PEDV reaches 99.6%. After 2 hours in simulated gastric fluid, L100@Al/Mn-MOF@PEDV nanoparticles maintained high stability with only about 12.73% release rate of inactivated PEDV. Uptake experiments in mouse RAW264.7 cells show enhanced uptake of Al/Mn-MOF@PEDV compared to unencapsulated inactivated PEDV. Additionally, Al/Mn-MOF@PEDV significantly boosts the expression of Th1-type factor IFN-γ and Th2-type factor IL-4 in RAW264.7 cells. This research offers new insights into the design of oral carriers for inactivated PED vaccines and lays the foundation for future applications in animal immunization.

Key words: PEDV, inactivated vaccine, nanoparticle carrier, metal-organic framework, oral delivery

CLC Number:

S855.3

HU Mi, SHEN Yaoxin, FAN Baochao, SUN Min, ZHOU Jinzhu, GUO Rongli, LI Bin. Evaluation of the Characteristics of Eudragit L100-Modified Aluminum-Manganese Metal-Organic Framework as an Oral Delivery Vehicle for Inactivated Porcine Epidemic Diarrhea Vaccine[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(5): 2292-2230.

Figures/Tables 7

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References 25

1	ZHANGH,ZOUC C,PENGO Y,et al.Global dynamics of porcine enteric coronavirus PEDV epidemiology, evolution, and transmission[J].Mol Biol Evol,2023,40(3):msad052. doi: 10.1093/molbev/msad052
2	SONGD,MOONH,KANGB.Porcine epidemic diarrhea: a review of current epidemiology and available vaccines[J].Clin Exp Vaccine Res,2015,4(2):166-176. doi: 10.7774/cevr.2015.4.2.166
3	NIUX Y,WANGQ H.Prevention and control of porcine epidemic diarrhea: the development of recombination-resistant live attenuated vaccines[J].Viruses (Basel),2022,14(6):1317. doi: 10.3390/v14061317
4	WANGQ H,VLASOVAA N,KENNEYS P,et al.Emerging and re-emerging coronaviruses in pigs[J].Curr Opin Virol,2019,34,39-49. doi: 10.1016/j.coviro.2018.12.001
5	李天芝,于新友,沈志强.黏膜免疫及其在猪病防控中的应用[J].猪业科学,2019,36(3):90-94.
	LIT Z,YUX Y,SHENZ Q.Mucosal immunity and its application in prevention and control of swine diseases[J].Swine Industry Science,2019,36(3):90-94.
6	QINZ L,NAIZ D,LIG,et al.The oral inactivated porcine epidemic diarrhea virus presenting in the intestine induces mucosal immunity in mice with alginate-chitosan microcapsules[J].Animals (Basel),2023,13(5):889.
7	MIAOY B,PANW Y,CHENK H,et al.Engineering a nanoscale Al-MOF-armored antigen carried by a "trojan horse"-like platform for oral vaccination to induce potent and long-lasting immunity[J].Adv Funct Mater,2019,29(43):1904828. doi: 10.1002/adfm.201904828
8	ZHONGX F,ZHANGY T,TANL,et al.An aluminum adjuvant-integrated nano-MOF as antigen delivery system to induce strong humoral and cellular immune responses[J].J Control Release,2019,300,81-92. doi: 10.1016/j.jconrel.2019.02.035
9	MARRACKP,MCKEEA S,MUNKSM W.Towards an understanding of the adjuvant action of aluminium[J].Nat Rev Immunol,2009,9(4):287-293. doi: 10.1038/nri2510
10	WANGZ M,YUANY M,CHENC,et al.Colloidal manganese salt improves the efficacy of rabies vaccines in mice, cats, and dogs[J].J Virol,2021,95(23):e0141421. doi: 10.1128/JVI.01414-21
11	THAKRALS,THAKRALN K,MAJUMDARD K.Eudragit^®: a technology evaluation[J].Expert Opin Drug Deliv,2013,10(1):131-149. doi: 10.1517/17425247.2013.736962
12	FANB C,SUNJ,ZHUL,et al.Development of a novel double antibody sandwich quantitative enzyme-linked immunosorbent assay for detection of porcine epidemic diarrhea virus antigen[J].Front Vet Sci,2020,7,540248. doi: 10.3389/fvets.2020.540248
13	FARZANEHF,HAMIDIPOURL.Mn-metal organic framework as heterogenous catalyst for oxidation of alkanes and alkenes[J].J Sci Islamic Repub Iran,2016,27(1):31-37.
14	HAOS L,WANGB C,WANGY Z,et al.Preparation of Eudragit L 100-55 enteric nanoparticles by a novel emulsion diffusion method[J].Colloids Surf B: Biointerfaces,2013,108,127-133. doi: 10.1016/j.colsurfb.2013.02.036
15	LUANY,QIY,GAOH Y,et al.A general post-synthetic modification approach of amino-tagged metal-organic frameworks to access efficient catalysts for the Knoevenagel condensation reaction[J].J Mater Chem A,2015,3(33):17320-17331. doi: 10.1039/C5TA00816F
16	XIAOY,WANGX N,LIY,et al.Evaluation of the immunogenicity in mice orally immunized with recombinant Lactobacillus casei expressing porcine epidemic diarrhea virus S1 protein[J].Viruses (Basel),2022,14(5):890. doi: 10.3390/v14050890
17	HUANGK Y,YANGG L,JINY B,et al.Construction and immunogenicity analysis of Lactobacillus plantarum expressing a porcine epidemic diarrhea virus S gene fused to a DC-targeting peptide[J].Virus Res,2018,247,84-93. doi: 10.1016/j.virusres.2017.12.011
18	WANGX L,WANGZ B,XUH Y,et al.Orally administrated whole yeast vaccine against porcine epidemic diarrhea virus induced high levels of IgA response in mice and piglets[J].Viral Immunol,2016,29(9):526-531. doi: 10.1089/vim.2016.0067
19	WANGF Q,ZHANGQ,ZHANGF Q,et al.Adenovirus vector-mediated single chain variable fragments target the nucleocapsid protein of porcine epidemic diarrhea virus and protect against viral infection in piglets[J].Front Immunol,2023,14,1058327. doi: 10.3389/fimmu.2023.1058327
20	DINGC C,MAJ F,DONGQ L,et al.Live bacterial vaccine vector and delivery strategies of heterologous antigen: a review[J].Immunol Lett,2018,197,70-77. doi: 10.1016/j.imlet.2018.03.006
21	LEEW J,CHAS,SHINM,et al.Efficacy of thiolated eudragit microspheres as an oral vaccine delivery system to induce mucosal immunity against enterotoxigenic Escherichia coli in mice[J].Eur J Pharm Biopharm,2012,81(1):43-48. doi: 10.1016/j.ejpb.2012.01.010
22	HORIM,ONISHIH,MACHIDAY.Evaluation of Eudragit-coated chitosan microparticles as an oral immune delivery system[J].Int J Pharm,2005,297(1-2):223-234. doi: 10.1016/j.ijpharm.2005.04.008
23	CURRYH,ALVAREZG R,ZWILLINGB S,et al.Toll-like receptor 2 stimulation decreases IFN-γ receptor expression in mouse RAW264. 7 macrophages[J].J Interf Cytok Res,2004,24(12):699-710. doi: 10.1089/jir.2004.24.699
24	GORDONS.Alternative activation of macrophages[J].Nat Rev Immunol,2003,3(1):23-35. doi: 10.1038/nri978
25	CENY W,CHENS J,WEIS Y,et al.A unique combination of Mn²⁺ and aluminum adjuvant acted the synergistic effect[J].Can J Infect Dis Med Microbiol,2024,2024,7502110.

试验方案 Protocol	灭活PEDV/金属离子/有机配体投料比 Feeding ratio of PEDV/Metal ion/Organic linker	负载效率/% Loading efficiency (LE)
1	0.42∶1.0∶1.0	56.3
2	0.21∶1.0∶1.0	85.7
3	0.14∶1.0∶1.0	99.6

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Evaluation of the Characteristics of Eudragit L100-Modified Aluminum-Manganese Metal-Organic Framework as an Oral Delivery Vehicle for Inactivated Porcine Epidemic Diarrhea Vaccine

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