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

Research Paper

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.

Key words

chemical modification immune response konjac glucomannan nanoparticles vaccine adjuvants 

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

Notes

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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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.State Key Laboratory of Agricultural Microbiology, College of ScienceHuazhong Agricultural UniversityWuhanPeople’s Republic of China

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