Back to Journals » International Journal of Nanomedicine » Volume 7

Positively charged cholesterol–recombinant human gelatins foster the cellular uptake of proteins and murine immune reactions

Authors Kadengodlu PA, Hebishima T, Takeshima SN, Ito M, Liu M, Abe H, Aida Y, Aigaki T, Ito Y

Received 25 July 2012

Accepted for publication 17 August 2012

Published 11 October 2012 Volume 2012:7 Pages 5437—5450

DOI https://doi.org/10.2147/IJN.S36350

Checked for plagiarism Yes

Review by Single-blind

Peer reviewer comments 2

Pallavi A Kadengodlu,1,3 Takehisa Hebishima,2 Shin-Nosuke Takeshima,2 Mika Ito,1 Mingzhe Liu,1 Hiroshi Abe,1 Yoko Aida,2 Toshiro Aigaki,3 Yoshihiro Ito1,3

1
Nano Medical Engineering Laboratory, 2Viral Infectious Diseases Unit, RIKEN Advance Science Institute, Wako, Saitama, Japan; 3Graduate School of Biological Science, Tokyo Metropolitan University, Tokyo, Japan

Purpose: Recombinant human gelatins with defined molecular weights were modified with cholesterol to make them amphiphilic in nature. We investigated the feasibility of these modified human gelatins acting as a carrier of antigenic proteins for inducing cellular immunity. The aim of this study was to synthesize novel and effective compounds for vaccine delivery in vivo.
Methods: Two types of cholesterol-modified gelatin micelles, anionic cholesterol-modified gelatin (aCMG) and cationic-cholesterol modified gelatin (cCMG), were synthesized using different cholesterol derivatives such as the cholesterol-isocyanate (Ch-I) for aCMG and amino-modified cholesterol for cCMG. One was anionic and the other cationic, and therefore they differed in terms of their zeta potential. The aCMG and cCMG were characterized for their size, zeta potential, and in their ability to form micelles. Cytotoxicity was also evaluated. The modified human gelatins were then investigated as a carrier of antigenic proteins for inducing cellular immunity both in vitro in DC 2.4 cells, a murine dendritic cell line, as well as in vivo. The mechanism of entry of the polymeric micelles into the cells was also evaluated.
Results: It was found that only cCMG successfully complexed with the model antigenic protein, fluorescein-isothiocyanate ovalbumin (OVA) and efficiently delivered and processed proteins in DC 2.4 cells. It was hypothesized that cCMG enter the cells predominantly by a caveolae-mediated pathway that required tyrosine kinase receptors on the cell surface. Animal testing using mice showed that the cationic cholesterol-modified gelatin complexed with OVA produced significantly high antibody titers against OVA: 2580-fold higher than in mice immunized with free OVA.
Conclusion: Conclusively, cCMG has shown to be very effective in stimulating an immune response due to its high efficiency, stability, and negligible cytotoxicity.

Keywords: recombinant human gelatin, cholesterol, micelle, protein delivery, caveolae pathway, receptor-mediated endocytosis

Creative Commons License This work is published and licensed by Dove Medical Press Limited. The full terms of this license are available at https://www.dovepress.com/terms.php and incorporate the Creative Commons Attribution - Non Commercial (unported, v3.0) License. By accessing the work you hereby accept the Terms. Non-commercial uses of the work are permitted without any further permission from Dove Medical Press Limited, provided the work is properly attributed. For permission for commercial use of this work, please see paragraphs 4.2 and 5 of our Terms.

Download Article [PDF]  View Full Text [HTML][Machine readable]

 

Other articles by this author:

Nanolayer formation on titanium by phosphonated gelatin for cell adhesion and growth enhancement

Zhou X, Park SH, Mao H, Isoshima T, Wang Y, Ito Y

International Journal of Nanomedicine 2015, 10:5597-5607

Published Date: 2 September 2015

Mussel-inspired human gelatin nanocoating for creating biologically adhesive surfaces

Yang X, Zhu L, Tada S, Zhou D, Kitajima T, Isoshima T, Yoshida Y, Nakamura M, Yan W, Ito Y

International Journal of Nanomedicine 2014, 9:2753-2765

Published Date: 29 May 2014

Readers of this article also read:

Novel PLGA-based nanoparticles for the oral delivery of insulin

Malathi S, Nandhakumar P, Pandiyan V, Webster TJ, Balasubramanian S

International Journal of Nanomedicine 2015, 10:2207-2218

Published Date: 19 March 2015

Green synthesis of water-soluble nontoxic polymeric nanocomposites containing silver nanoparticles

Prozorova GF, Pozdnyakov AS, Kuznetsova NP, Korzhova SA, Emel’yanov AI, Ermakova TG, Fadeeva TV, Sosedova LM

International Journal of Nanomedicine 2014, 9:1883-1889

Published Date: 16 April 2014

Methacrylic-based nanogels for the pH-sensitive delivery of 5-Fluorouracil in the colon

Ashwanikumar N, Kumar NA, Nair SA, Kumar GS

International Journal of Nanomedicine 2012, 7:5769-5779

Published Date: 15 November 2012

A novel preparation method for silicone oil nanoemulsions and its application for coating hair with silicone

Hu Z, Liao M, Chen Y, Cai Y, Meng L, Liu Y, Lv N, Liu Z, Yuan W

International Journal of Nanomedicine 2012, 7:5719-5724

Published Date: 12 November 2012

Cross-linked acrylic hydrogel for the controlled delivery of hydrophobic drugs in cancer therapy

Deepa G, Thulasidasan AK, Anto RJ, Pillai JJ, Kumar GS

International Journal of Nanomedicine 2012, 7:4077-4088

Published Date: 27 July 2012

Corrigendum

Chen ZQ, Liu Y, Zhao JH, Wang L, Feng NP

International Journal of Nanomedicine 2012, 7:1709-1710

Published Date: 30 March 2012

Crystallization after intravitreal ganciclovir injection

Pitipol Choopong, Nattaporn Tesavibul, Nattawut Rodanant

Clinical Ophthalmology 2010, 4:709-711

Published Date: 14 July 2010