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

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

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 Ito1,31Nano Medical Engineering Laboratory, 2Viral Infectious Diseases Unit, RIKEN Advance Science Institute, Wako, Saitama, Japan; 3Graduate School of...

Descripción completa

Guardado en:
Detalles Bibliográficos
Autores principales: Kadengodlu PA, Hebishima T, Takeshima SN, Ito M, Liu M, Abe H, Aida Y, Aigaki T, Ito Y
Formato: article
Lenguaje:EN
Publicado: Dove Medical Press 2012
Materias:
Medicine (General)
R5-920
Acceso en línea:https://doaj.org/article/9ef8895996c14f279b5384751a402ce6
Etiquetas: Agregar Etiqueta
Sin Etiquetas, Sea el primero en etiquetar este registro!
id oai:doaj.org-article:9ef8895996c14f279b5384751a402ce6
record_format dspace
spelling oai:doaj.org-article:9ef8895996c14f279b5384751a402ce62021-12-02T02:28:10ZPositively charged cholesterol–recombinant human gelatins foster the cellular uptake of proteins and murine immune reactions1176-91141178-2013https://doaj.org/article/9ef8895996c14f279b5384751a402ce62012-10-01T00:00:00Zhttp://www.dovepress.com/positively-charged-cholesterolndashrecombinant-human-gelatins-foster-t-a11249https://doaj.org/toc/1176-9114https://doaj.org/toc/1178-2013Pallavi 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 Ito1,31Nano Medical Engineering Laboratory, 2Viral Infectious Diseases Unit, RIKEN Advance Science Institute, Wako, Saitama, Japan; 3Graduate School of Biological Science, Tokyo Metropolitan University, Tokyo, JapanPurpose: 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 endocytosisKadengodlu PAHebishima TTakeshima SNIto MLiu MAbe HAida YAigaki TIto YDove Medical PressarticleMedicine (General)R5-920ENInternational Journal of Nanomedicine, Vol 2012, Iss default, Pp 5437-5450 (2012)
institution DOAJ
collection DOAJ
language EN
topic Medicine (General)
R5-920
spellingShingle Medicine (General)
R5-920
Kadengodlu PA
Hebishima T
Takeshima SN
Ito M
Liu M
Abe H
Aida Y
Aigaki T
Ito Y
Positively charged cholesterol–recombinant human gelatins foster the cellular uptake of proteins and murine immune reactions
description 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 Ito1,31Nano Medical Engineering Laboratory, 2Viral Infectious Diseases Unit, RIKEN Advance Science Institute, Wako, Saitama, Japan; 3Graduate School of Biological Science, Tokyo Metropolitan University, Tokyo, JapanPurpose: 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
format article
author Kadengodlu PA
Hebishima T
Takeshima SN
Ito M
Liu M
Abe H
Aida Y
Aigaki T
Ito Y
author_facet Kadengodlu PA
Hebishima T
Takeshima SN
Ito M
Liu M
Abe H
Aida Y
Aigaki T
Ito Y
author_sort Kadengodlu PA
title Positively charged cholesterol–recombinant human gelatins foster the cellular uptake of proteins and murine immune reactions
title_short Positively charged cholesterol–recombinant human gelatins foster the cellular uptake of proteins and murine immune reactions
title_full Positively charged cholesterol–recombinant human gelatins foster the cellular uptake of proteins and murine immune reactions
title_fullStr Positively charged cholesterol–recombinant human gelatins foster the cellular uptake of proteins and murine immune reactions
title_full_unstemmed Positively charged cholesterol–recombinant human gelatins foster the cellular uptake of proteins and murine immune reactions
title_sort positively charged cholesterol–recombinant human gelatins foster the cellular uptake of proteins and murine immune reactions
publisher Dove Medical Press
publishDate 2012
url https://doaj.org/article/9ef8895996c14f279b5384751a402ce6
work_keys_str_mv AT kadengodlupa positivelychargedcholesterolampndashrecombinanthumangelatinsfosterthecellularuptakeofproteinsandmurineimmunereactions
AT hebishimat positivelychargedcholesterolampndashrecombinanthumangelatinsfosterthecellularuptakeofproteinsandmurineimmunereactions
AT takeshimasn positivelychargedcholesterolampndashrecombinanthumangelatinsfosterthecellularuptakeofproteinsandmurineimmunereactions
AT itom positivelychargedcholesterolampndashrecombinanthumangelatinsfosterthecellularuptakeofproteinsandmurineimmunereactions
AT lium positivelychargedcholesterolampndashrecombinanthumangelatinsfosterthecellularuptakeofproteinsandmurineimmunereactions
AT abeh positivelychargedcholesterolampndashrecombinanthumangelatinsfosterthecellularuptakeofproteinsandmurineimmunereactions
AT aiday positivelychargedcholesterolampndashrecombinanthumangelatinsfosterthecellularuptakeofproteinsandmurineimmunereactions
AT aigakit positivelychargedcholesterolampndashrecombinanthumangelatinsfosterthecellularuptakeofproteinsandmurineimmunereactions
AT itoy positivelychargedcholesterolampndashrecombinanthumangelatinsfosterthecellularuptakeofproteinsandmurineimmunereactions
_version_ 1718402487427268608

Ejemplares similares