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  1. Article ; Online: Design of a lipid nano-delivery system containing recombinant Candida albicans chitinase 3 as a potential vaccine against fungal infections.

    Costa-Barbosa, Augusto / Pacheco, Maria Inês / Carneiro, Catarina / Botelho, Cláudia / Gomes, Andreia C / Real Oliveira, M Elisabete C D / Collins, Tony / Vilanova, Manuel / Pais, Célia / Correia, Alexandra / Sampaio, Paula

    Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

    2023  Volume 166, Page(s) 115362

    Abstract: Opportunistic fungi cause lethal systemic infections and impose high medical costs to health systems. The World Health Organization has recognized the importance of fungal infections, including them in its global priority list guiding research, ... ...

    Abstract Opportunistic fungi cause lethal systemic infections and impose high medical costs to health systems. The World Health Organization has recognized the importance of fungal infections, including them in its global priority list guiding research, development, and discovery of new therapeutic approaches. Fungal vaccine development has been proposed as one of the treatment and prevention strategies in the last decade. In this study, we present the design of a lipid antigen delivery system based on Dioctadecyldimethylammonium bromide: Monoolein (DODAB: MO) containing recombinant Candida albicans Chitinase 3 (Cht3) for modulation the immune response against fungal infections. Several DODAB:MO liposomes containing Cht3 were prepared and those prepared by the incubation method and containing 5 µg/mL Cht3 were selected due to their favorable size, ζ-potential and stability, suited for antigen delivery applications. The encapsulation of Cht3 in these liposomes resulted in a significant increase in cellular uptake compared to empty liposomes, demonstrating their efficacy in delivering the antigen. Moreover, the liposomes proved to be safe for use in immunization procedures. Subcutaneous administration of Cht3 liposomes elicited a Th1/Th17 immune response profile, associated with the production of high levels of antibodies against Cht3. These antibodies recognized both the native and the recombinant forms of the protein, opsonizing mother-yeast at the cell scars, which has the potential to disrupt cell separation and hinder yeast growth. The findings suggest that the designed lipid antigen delivery system shows promise as a potential candidate for enhancing immune responses against fungal infections, offering a valuable strategy for future fungal vaccine development.
    MeSH term(s) Candida albicans ; Chitinases ; Liposomes ; Vaccines ; Fungal Vaccines ; Mycoses ; Antibodies ; Lipids
    Chemical Substances Chitinases (EC 3.2.1.14) ; dimethyldioctadecylammonium (251IW5I21C) ; Liposomes ; Vaccines ; Fungal Vaccines ; Antibodies ; Lipids
    Language English
    Publishing date 2023-08-24
    Publishing country France
    Document type Journal Article
    ZDB-ID 392415-4
    ISSN 1950-6007 ; 0753-3322 ; 0300-0893
    ISSN (online) 1950-6007
    ISSN 0753-3322 ; 0300-0893
    DOI 10.1016/j.biopha.2023.115362
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Candida albicans chitinase 3 with potential as a vaccine antigen: production, purification, and characterisation.

    Costa-Barbosa, Augusto / Ferreira, Diogo / Pacheco, Maria Inês / Casal, Margarida / Duarte, Henrique Oliveira / Gomes, Catarina / Barbosa, Ana Margarida / Torrado, Egídio / Sampaio, Paula / Collins, Tony

    Biotechnology journal

    2023  Volume 19, Issue 1, Page(s) e2300219

    Abstract: Chitinases are widely studied enzymes that have already found widespread application. Their continued development and valorisation will be driven by the identification of new and improved variants and/or novel applications bringing benefits to industry ... ...

    Abstract Chitinases are widely studied enzymes that have already found widespread application. Their continued development and valorisation will be driven by the identification of new and improved variants and/or novel applications bringing benefits to industry and society. We previously identified a novel application for chitinases wherein the Candida albicans cell wall surface chitinase 3 (Cht3) was shown to have potential in vaccine applications as a subunit antigen against fungal infections. In the present study, this enzyme was investigated further, developing production and purification protocols, enriching our understanding of its properties, and advancing its application potential. Cht3 was heterologously expressed in Pichia pastoris and a 4-step purification protocol developed and optimised: this involves activated carbon treatment, hydrophobic interaction chromatography, ammonium sulphate precipitation, and gel filtration chromatography. The recombinant enzyme was shown to be mainly O-glycosylated and to retain the epitopes of the native protein. Functional studies showed it to be highly specific, displaying activity on chitin, chitosan, and chito-oligosaccharides larger than chitotriose only. Furthermore, it was shown to be a stable enzyme, exhibiting activity, and stability over broad pH and temperature ranges. This study represents an important step forward in our understanding of Cht3 and contributes to its development for application.
    MeSH term(s) Candida albicans/genetics ; Candida albicans/metabolism ; Chitinases/genetics ; Chitinases/chemistry ; Proteins ; Chitin/chemistry ; Chitin/metabolism ; Chitosan ; Hydrogen-Ion Concentration
    Chemical Substances Chitinases (EC 3.2.1.14) ; Proteins ; Chitin (1398-61-4) ; Chitosan (9012-76-4)
    Language English
    Publishing date 2023-10-30
    Publishing country Germany
    Document type Journal Article
    ZDB-ID 2221885-3
    ISSN 1860-7314 ; 1860-6768
    ISSN (online) 1860-7314
    ISSN 1860-6768
    DOI 10.1002/biot.202300219
    Database MEDical Literature Analysis and Retrieval System OnLINE

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