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Preparation of Modified Konjac Glucomannan Nanoparticles and their Application as Vaccine Adjuvants to Promote Ovalbumin-Induced Immune Response in Mice

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Abstract

Purpose

Herein, we reported a facile strategy for synthesis of two types of modified konjac glucomannan nanoparticles (NPs). The goal of this project was to explore the potential of the NPs as vaccine adjuvants.

Methods

Firstly, anionic carboxymethylated konjac glucomannan (CKGM) and cationic quaternized konjac glucomannan (QKGM) were synthesized by chemical modification of konjac glucomannan (KGM). Subsequently, two types of NPs, CKGM/QKGM and sodium tripolyphosphate (TPP)/QKGM, were prepared through polyelectrolyte complex method and ionic cross-linking method, respectively. The thus-synthesized NPs were then loaded with ovalbumin (OVA) to further evaluate the effect of NPs on immune response in mice.

Results

The encapsulation efficiency of OVA for CKGM/QKGM/OVA and TPP/QKGM/OVA NPs could be 49.2% and 67.7%, respectively, while the drug loading capacity could reach 10.9% and 60%. The NPs showed irregular spherical shape and exhibited good sustained-release properties. In vitro cytotoxicity assay revealed that both the blank and OVA-loaded NPs were not toxic to cells. The OVA-specific IgG, splenocytes proliferation and cytokine levels indicated that the OVA-induced humoral and cellular immune responses were up-regulated by OVA-loaded NPs. What’s more, CKGM/QKGM/OVA NPs elicited both higher IL-2 and IFN-γ production, while TPP/QKGM/OVA NPs elicited both higher IL-4 and IL-10 production.

Conclusions

These results suggest that TPP/QKGM and CKGM/QKGM NPs are promising to be used as vaccine adjuvants. The TPP/QKGM/OVA NPs could induce stronger humoral immune response, while CKGM/QKGM/OVA NPs could enhance the cellular immune response more effectively.

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Abbreviations

AHKGM:

Acid-hydrolysed konjac glucomannan

CCK-8:

Cell counting kit-8

CKGM:

Carboxymethylated konjac glucomannan

DMEM:

Dulbecco’s modified Eagle’s medium

EE:

Encapsulation efficiency

ELISA:

Enzyme linked immunosorbent assay

FBS:

Fetal bovine serum

FT-IR:

Fourier transform infrared

IFN:

Interferon

IL:

Interleukin

KGM:

Konjac glucomannan

LC:

Loading capacity

MTT:

3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide

NMR:

Nuclear magnetic resonance

NPs:

Nanoparticles

OD:

Optical density

OVA:

Ovalbumin

PBS:

Phosphate buffer saline

QKGM:

Quaternized konjac glucomannan

RPMI:

Roswell Park Memorial Institute

RT:

Room temperature

SEM:

Standard error mean

SI:

Stimulation index

TEM:

Transmission electron microscope

TPP:

Sodium tripolyphosphate

TMB:

3, 3′, 5, 5’-Tetramethylbenzidine

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Acknowledgments and Disclosures

This work was financially supported by the Da BeiNong Group Promoted Project for Young Scholar of HZAU (Grant No. 2017DBN010), the open funds of the State Key Laboratory of Agricultural Microbiology (Grant No. AMLKF201507), the National Natural Science Foundation of China (Grant No. 21503085), and the Natural Science Foundation of Hubei Province (Grant No. 2015CFB233).

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Author notes

  1. Na Chen and Pei Zhu contributed equally to this work.

Authors and Affiliations

  1. State Key Laboratory of Agricultural Microbiology, College of Science, Huazhong Agricultural University, Wuhan, 430070, People’s Republic of China

    Na Chen, Pei Zhu, Ting Du, Kai Han, Dang Wang, Jianfeng Ye, Shaobo Xiao, Xiaozhou Ye & Yun Wang

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  1. Na Chen
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Correspondence to Shaobo Xiao or Xiaozhou Ye.

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Chen, N., Zhu, P., Du, T. et al. Preparation of Modified Konjac Glucomannan Nanoparticles and their Application as Vaccine Adjuvants to Promote Ovalbumin-Induced Immune Response in Mice. Pharm Res 35, 105 (2018). https://doi.org/10.1007/s11095-018-2381-x

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