Improved stability and immunological potential of tetanus toxoid containing surface engineered bilosomes following oral administration

doi:10.1016/j.nano.2013.08.012

Nanomedicine: Nanotechnology, Biology and Medicine

Volume 10, Issue 2, February 2014, Pages 431-440

https://doi.org/10.1016/j.nano.2013.08.012 Get rights and content

Abstract

The present study was designed with the objective to investigate the stability and potential of glucomannan-modified bilosomes (GM-bilosomes) in eliciting immune response following oral administration. GM-bilosomes exhibited desired quality attributes simultaneously maintaining the chemical and conformation stability of the tetanus toxoid (TT) entrapped in to freeze dried formulations. The GM-bilosomes exhibited excellent stability in different simulated biological fluids and sustained release profile up to 24 h. GM-bilosomes elicited significantly higher (P < 0.05) systemic immune response (serum IgG level) as compared to bilosomes, niosomes and alum adsorbed TT administered through oral route. More importantly, GM-bilosomes were found capable of inducing mucosal immune response, i.e. sIgA titre in salivary and intestinal secretions as well as cell mediated immune response (IL-2 and IFN-γ levels in spleen homogenate) which was not induced by i.m. TT, the conventional route of immunization. Conclusively, GM-bilosomes could be considered as a promising carrier and adjuvant system for oral mucosal immunization.

From the Clinical Editor

This team reports on the development and effects of a glucomannan-modified bilosome as an oral vaccine vector, using tetanus toxoid in the experiments. These GM-bilosomes not only elicited significantly higher systemic immune response as compared to bilosomes, niosomes and alum adsorbed orally administered TT, but also demonstrated mucosal immune response induction as well as cell mediated immune responses, which were not induced by the conventional route of immunization.

Graphical Abstract

GM-bilosomes exhibited significantly higher systemic immune response simultaneously maintaining the chemical and conformation stability of the tetanus toxoid (TT) entrapped in to freeze dried formulations and can be considered as a promising carrier and adjuvant system for oral mucosal immunization.

Section snippets

Materials and reagents

TT (1850 Lf/mL, Batch No. TAM 046) was provided as generous gift sample by Panacea Biotec, India. Tetanus Toxoid (Adsorbed I.P.) (Serum Institute of India Ltd., Batch No. 017I1017B) was purchased from local market. Secondary antibodies namely anti-mouse IgA (α-chain specific) peroxidase conjugate, anti-mouse IgG (γ-chain specific) peroxidase conjugate, 3,3′,5,5′-tetramethylbenzidine (TMB), Nunc Immuno™ Maxisorb F96 well solid plates, Cholesterol (CH), sodium deoxycholate (Na-DOC), Span 80,

Preparation and optimization of vesicular formulations

The size, PDI, zeta potential and entrapment efficiency (EE) of different vesicular formulations are shown in Table 1. Morphology determination by TEM analysis (Figure 1) further confirmed the formation of vesicles of almost spherical shape with smooth surface. The results obtained in TEM analysis were also in good correlation with the results of DLS analysis.

Lyophilization

All vesicular formulations were lyophilized in order to impart long term stability. A number of cryoprotectants (Table 2) were screened

Discussion

In the present report mannosylated bilosomes were proposed for selective uptake and delivery of TT to APCs through the oral route. GM has been reported to exhibit excellent stability against digestive enzymes19, 20 which can also increase the stability of bilosomes in GIT. Furthermore, polymeric nature can provide higher density of mannose molecules over the bilosomes surface which can result in more precise recognition and binding to mannose receptors over expressed on APCs. Therefore, we

Acknowledgment

Authors are thankful to Prof. KB Tikoo and Mr. Vinod Kumar for providing TEM facilities at NIPER. Authors appreciate the technical assistance provided by Mr. Rahul Mahajan.

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: Conflict of Interest and Disclosure: The authors report no financial interest that might pose a potential, perceived, or real conflict of interest.

: All sources of support for research: Authors are grateful to the Department of Biotechnology (DBT), Government of India, New Delhi, India for providing support.

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Research ArticleImproved stability and immunological potential of tetanus toxoid containing surface engineered bilosomes following oral administration

Abstract

From the Clinical Editor

Graphical Abstract

Section snippets

Materials and reagents

Preparation and optimization of vesicular formulations

Lyophilization

Discussion

Acknowledgment

Mannosylated niosomes as adjuvant-carrier system for oral genetic immunization against hepatitis B

Immunol Lett

Mannosylated niosomes as adjuvant-carrier system for oral mucosal immunization

J Liposome Res

Lymphocyte migration in the intestinal mucosa: entry, transit and emigration of lymphoid cells and the influence of antigen

Vet Immunol Immunopathol

Nanoparticles as potential oral delivery systems of proteins and vaccines: a mechanistic approach

J Control Release

Mucosal vaccines: the promise and the challenge

Nat Rev Immunol

Liposome-based delivery system for vaccine candidates: constructing an effective formulation

Nanomedicine

Non-ionic surfactant based vesicles (niosomes) in drug delivery

Int J Pharm

Chitosan microparticles for oral vaccination: preparation, characterization and preliminary in vivo uptake studies in murine Peyer's patches

Biomaterials

Preparation of coated nanoparticles for a new mucosal vaccine delivery system

Int J Pharm

Oral delivery of tetanus toxoid using vesicles containing bile salts (bilosomes) induces significant systemic and mucosal immunity

Methods

Advances in lipid-based subunit vaccine formulations

Curr Immunol Rev

Topical and mucosal liposomes for vaccine delivery

Wiley Interdiscip Rev Nanomed Nanobiotechnol

Cholera toxin B subunit conjugated bile salt stabilized vesicles (bilosomes) for oral immunization

Int J Pharm

Role of the glycocalyx in regulating access of microparticles to apical plasma membranes of intestinal epithelial cells: implications for microbial attachment and oral vaccine targeting

J Exp Med

The uptake and translocation of latex nanospheres and microspheres after oral administration to rats

J Pharm Pharmacol

Targeting to macrophages: role of physicochemical properties of particulate carriers-liposomes and microspheres-on the phagocytosis by macrophages

J Control Release

Development of phosphorylated glucomannan-coated chitosan nanoparticles as nanocarriers for protein delivery

J Nanosci Nanotechnol

Formation of new glucomannan-chitosan nanoparticles and study of their ability to associate and deliver proteins

Macromolecules

Research Article
Improved stability and immunological potential of tetanus toxoid containing surface engineered bilosomes following oral administration