Autor(es):
Rodrigues, Daniel Barreira ; Moreira, Helena R. ; Cerqueira, Mariana T. ; Marques, A. P. ; Castro, António G. ; Reis, R. L. ; Pirraco, Rogério P.
Data: 2022
Identificador Persistente: https://hdl.handle.net/1822/88570
Origem: RepositóriUM - Universidade do Minho
Projeto/bolsa:
info:eu-repo/grantAgreement/EC/H2020/805411/EU;
info:eu-repo/grantAgreement/FCT/POR_NORTE/SFRH%2FBD%2F119756%2F2016/PT;
info:eu-repo/grantAgreement/FCT/Investigador FCT/IF%2F00347%2F2015%2FCP1294%2FCT0004/PT;
Assunto(s): Cytotoxic T cells; Gellan Gum; Nanoparticles; T cell stimulation; Science & Technology
Descrição
Background: T cell priming has been shown to be a powerful immunotherapeutic approach for cancer treatment in terms of efcacy and relatively weak side efects. Systems that optimize the stimulation of T cells to improve therapeutic efcacy are therefore in constant demand. A way to achieve this is through artifcial antigen presenting cells that are complexes between vehicles and key molecules that target relevant T cell subpopulations, eliciting antigenspecifc T cell priming. In such T cell activator systems, the vehicles chosen to deliver and present the key molecules to the targeted cell populations are of extreme importance. In this work, a new platform for the creation of T cell activator systems based on highly tailorable nanoparticles made from the natural polymer gellan gum (GG) was developed and validated. Methods: GG nanoparticles were produced by a water in oil emulsion procedure, and characterized by dynamic light scattering, high resolution scanning electronic microscopy and water uptake. Their biocompatibility with cultured cells was assessed by a metabolic activity assay. Surface functionalization was performed with anti-CD3/ CD28 antibodies via EDC/NHS or NeutrAvidin/Biotin linkage. Functionalized particles were tested for their capacity to stimulate CD4+ T cells and trigger T cell cytotoxic responses. Results: Nanoparticles were approximately 150 nm in size, with a stable structure and no detectable cytotoxicity. Water uptake originated a weight gain of up to 3200%. The functional antibodies did efciently bind to the nanoparticles, as confrmed by SDS-PAGE, which then targeted the desired CD4+ populations, as confrmed by confocal microscopy. The developed system presented a more sustained T cell activation over time when compared to commercial alternatives. Concurrently, the expression of higher levels of key cytotoxic pathway molecules granzyme B/perforin was induced, suggesting a greater cytotoxic potential for future application in adoptive cancer therapy. Conclusions: Our results show that GG nanoparticles were successfully used as a highly tailorable T cell activator system platform capable of T cell expansion and re-education.
Authors would like to acknowledge the fnancial support from the European Research Council through the Starting Grant “CapBed” (ERC-2018-STG-805411), FCT/MCTES (Fundação para a Ciência e a Tecnologia/ Ministério da Ciência, Tecnologia, e Ensino Superior) through the grants SFRH/BD/119756/2016 (D.B.R.) and IF/00347/2015 (R.P.P.) and to the FSE/POCH (Fundo Social Europeu através do Programa Operacional do Capital Humano) under the scope of the PD/169/2013, NORTE-08–5369-FSE-000037 (H.R.M.).
