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

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 t...

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Autores principales: Kadengodlu, Pallavi A, Hebishima, Takehisa, Takeshima, Shin-Nosuke, Ito, Mika, Liu, Mingzhe, Abe, Hiroshi, Aida, Yoko, Aigaki, Toshiro, Ito, Yoshihiro
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Dove Medical Press 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3471541/
https://www.ncbi.nlm.nih.gov/pubmed/23091385
http://dx.doi.org/10.2147/IJN.S36350
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author Kadengodlu, Pallavi A
Hebishima, Takehisa
Takeshima, Shin-Nosuke
Ito, Mika
Liu, Mingzhe
Abe, Hiroshi
Aida, Yoko
Aigaki, Toshiro
Ito, Yoshihiro
author_facet Kadengodlu, Pallavi A
Hebishima, Takehisa
Takeshima, Shin-Nosuke
Ito, Mika
Liu, Mingzhe
Abe, Hiroshi
Aida, Yoko
Aigaki, Toshiro
Ito, Yoshihiro
author_sort Kadengodlu, Pallavi A
collection PubMed
description 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.
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spelling pubmed-34715412012-10-22 Positively charged cholesterol–recombinant human gelatins foster the cellular uptake of proteins and murine immune reactions Kadengodlu, Pallavi A Hebishima, Takehisa Takeshima, Shin-Nosuke Ito, Mika Liu, Mingzhe Abe, Hiroshi Aida, Yoko Aigaki, Toshiro Ito, Yoshihiro Int J Nanomedicine Original Research 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. Dove Medical Press 2012 2012-10-11 /pmc/articles/PMC3471541/ /pubmed/23091385 http://dx.doi.org/10.2147/IJN.S36350 Text en © 2012 Kadengodlu et al, publisher and licensee Dove Medical Press Ltd. This is an Open Access article which permits unrestricted noncommercial use, provided the original work is properly cited.
spellingShingle Original Research
Kadengodlu, Pallavi A
Hebishima, Takehisa
Takeshima, Shin-Nosuke
Ito, Mika
Liu, Mingzhe
Abe, Hiroshi
Aida, Yoko
Aigaki, Toshiro
Ito, Yoshihiro
Positively charged cholesterol–recombinant human gelatins foster the cellular uptake of proteins and murine immune reactions
title 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_short 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
topic Original Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3471541/
https://www.ncbi.nlm.nih.gov/pubmed/23091385
http://dx.doi.org/10.2147/IJN.S36350
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