Carboxymethylation enhances the maturation-inducing activity in dendritic cells of polysaccharide from the seeds of Plantago asiatica L.

doi:10.1016/j.intimp.2014.06.027

International Immunopharmacology

Volume 22, Issue 2, October 2014, Pages 324-331

https://doi.org/10.1016/j.intimp.2014.06.027 Get rights and content

Highlights

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Carboxymethylated PLCP enhances the expression of surface molecules of DCs.
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Carboxymethylated PLCP enhances the stimulatory capacity of MLR of DCs.
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Carboxymethylated PLCP stimulates the secretion of IL-12p70 cytokine of DCs.
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Carboxymethylated PLCP induces the mRNA synthesis of chemokine receptors of DCs.
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Carboxymethylated polysaccharides can be used as immunotherapeutic adjuvant.

Abstract

Carboxymethylation is a well-known modification process for polysaccharides. To evaluate the biological availability of carboxymethyl, polysaccharide from the seeds of Plantago asiatica L. (PLCP) was carboxymethylated (CM-PLCP) and the immunomodulatory activities of five CM-PLCPs of gradient degree of substitution (DS) from 0.40 to 0.62 were determined on dendritic cells (DCs) in vitro. Compared with DCs treated with PLCP, DCs treated with CM-PLCP of DS_0.50, DS_0.55, DS_0.62, as well as CD86 and CD80, expressed higher levels of MHCII, CD86 and CD80 surface molecules. In addition, the secretion of IL-12p70 and the mRNA of CCR7 and CXCR4 chemokines were increased, while the endocytosis activities were inhibited. Correspondingly, stronger mixed lymphocyte reactions were induced by the DCs treated with the CM-PLCPs. The results showed that carboxymethylation modification of relevant high DS can enhance the DC maturation-inducing function of PLCP, indicating the potential application of carboxymethylated polysaccharide as an immunotherapeutic adjuvant.

Graphical abstract

Introduction

In the past decades, numerous studies come to the viewpoint that natural polysaccharides have a broad spectrum of immunological function. Recently, semi-synthetic polysaccharides and derivatives from natural polysaccharides prepared by chemical or enzymatic modification have been reported to exhibit high biological properties [1]. For the advantage of low cost of the chemicals and the non-toxicity of the products, carboxymethylation reactions are applied to natural polymers, such as chitosan [2], starch [3] and kappa-carrageenan [4], to obtain biomaterials in a wide variety of fields.

Plantaginaceae is the largest genus of the cosmopolitan family and some Plantago species are confirmed to have beneficial bioactivities, such as anti-inflammatory, anti-cancer and anti-oxidant activities [5], [6], [7], [8]. Plantago asiatica L., a traditional Chinese herbal medicine, is used in the treatment of many diseases. We have recently reported the structure and bioactivities of crude polysaccharide from the seeds of P. asiatica L. (PLCP). It contains three fractions, i.e., PLP-1, PLP-2 and PLP-3. The main fraction of PLP-2 is a highly branched heteroxylan which consisted of a β-1,4-linked Xylp backbone with side chains attached to O-2 or O-3 [9]. PLP-3 was found to be arabinoxylan which consisted of a β-1,4-linked Xylp backbone with short side chains attached to its O-2 (1,2,4-linked Xylp, 17.87%) or O-3 (1,3,4-linked Xylp, 24.24%) positions [10]. Interestingly, we found that PLCP could induce the maturation of dendritic cells (DCs) [11], which orchestrate the innate and adaptive immunity. So far, whether the carboxymethylation modification would influence the immunoregulation activity of PLCP is still unknown.

In the present study, the effect of five carboxymethylated PLCPs (CM-PLCPs) on DCs were determined. The main purpose was to determine whether carboxymethylation modification could raise the immune enhancement of PLCP.

Section snippets

Preparation of PLCP and CM-PLCPs

The seeds of P. asiatica L. were purchased from Ji'an County, Jiangxi Province, China, and air dried before use. The species were identified by Dr. Cui-sheng Fan, Jiangxi University of Traditional Chinese Medicine, Nanchang, China.

Dried seeds of P. asiatica L. were extracted three times with tenfold volumes of distilled water at 90 °C for 2 h. The combined supernatant was concentrated at 55 °C. A fourfold volume of ethanol was added to the concentrated solution to precipitate polysaccharide. The

FT-IR spectroscopy analysis

The FT-IR spectrum of PLCP and CM-PLCPs is showed in Fig. 1. The strong absorption at 3429.9 cm^− 1 was due to hydroxyl stretching vibration of the polysaccharide and the absorption at 2927.0 cm^− 1 was due to the C single bond H stretching vibration. The characteristic anomeric region absorption bands at 893.2 cm^− 1indicates a β configuration of the polysaccharide. The absorbance at 1729.2 cm^− 1 is characteristic of C double bond O stretching vibration of carboxylic acid. Compared with PLCP, CM-PLPs have a stronger absorption at

Discussion

Abundant polysaccharides obtained from plants, animals and microorganisms have long been believed to have immunological enhancement properties [19]. These bioactivities are based on their chemical structure and configuration, however, little is known about the relationship between structure and function. One approach to studying such structure–function relationship is to compare the properties of families of polysaccharides in which individual members of the group show small naturally arising

Conclusion

In the current study, we have demonstrated that relevant high carboxymethylation modification can enhance the DC maturation-inducing function of PLCP, indicating the potential application of carboxymethylated polysaccharide as an immunotherapeutic adjuvant.

Abbreviations

cluster of differentiation

carboxymethylated

DCs

dendritic cells

degree of substitution

FT-IR

Fourier transform infrared spectroscopy

PLCP

polysaccharide from the seeds of Plantago asiatica L.

RPMI

Roswell Park Memorial Institute

Acknowledgment

This study is supported by the National Key Technology R & D Program of China (No. 2012BAD33B06), the Key Program of National Natural Science Foundation of China (No: 31130041), the National Natural Science Foundation of China (No. 21062012), the National High-tech R & D (863 Program)(No. 2013AA102102), the Program for New Century Excellent Talents in University (NCET-12-0749), and the College Students Innovative Training Project of Nanchang University (No. 2012044), which are gratefully

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Carboxymethylation enhances the maturation-inducing activity in dendritic cells of polysaccharide from the seeds of Plantago asiatica L.

Highlights

Abstract

Graphical abstract

Introduction

Section snippets

Preparation of PLCP and CM-PLCPs

FT-IR spectroscopy analysis

Discussion

Conclusion

Abbreviations

Acknowledgment

Biomaterials

Int J Pharm

Carbohydr Polym

LWT – Food Sci Technol

J Ethnopharmacol

J Ethnopharmacol

Int J Biol Macromol

Carbohydr Polym

Carbohydr Polym

Eur J Pharmacol

Carbohydr Polym

Clin Appl Immunol Rev

Int J Biol Macromol

Carbohydr Polym

Carbohydr Res

Carbohydr Res

Curr Opin Immunol

Cell

Int J Biol Macromol

Int Immunopharmacol

Immunity