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Development of potent class II transactivator gene delivery systems capable of inducing de novo MHC II expression in human cells, in vitro and ex vivo

Gene Therapy volume 24, pages 342–352 (2017)Cite this article

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Abstract

Class II transactivator (CIITA) induces transcription of major histocompatibility complex (MHC) II genes and can potentially be used to improve genetic immunotherapies by converting non-immune cells into cells capable of presenting antigens to CD4+ T cells. However, CIITA expression is tightly controlled and it remains unclear whether distinct non-immune cells differ in this transactivator regulation. Here we describe the development of gene delivery systems capable of promoting the efficient CIITA expression in non-immune cell lines and in primary human cells of an ex vivo skin explant model. Different human cell types undergoing CIITA overexpression presented high-level de novo expression of MHC II, validating the delivery systems as suitable tools for the CIITA evaluation as a molecular adjuvant for gene therapies.

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Acknowledgements

This work was supported by the Fundação de Amparo a Pesquisa do Estado de São Paulo Grants 2011/20649-6, 2011/12134-6 and 2012/22171-9 and by the Fondazione Ri.MED. BD is a Fondazione Ri.MED Associate Scholar. We thank Stefan P Lamers for manuscript editing. We also acknowledge Dr Patrícia A Toniolo and Dr Elizabeth R Wonderlich for assistance with the lentivirus transduction system, and Dr Jared Evans for kindly providing the pcDNAl-eGFP plasmid.

Author contributions

MLP designed, performed and analyzed most of the experiments, and wrote the manuscript. PD performed the skin explant inoculations, immunohistochemistry, acquisition of confocal images and the density of markers quantification in confocal images. BD designed, performed and analyzed the quantitative real-time PCR assays. IFTV provided technical assistance. RD performed the artificial sequence design and optimization. POR provided technical assistance. EJMN, TMO, RBM, SMBB, AJSD and ETAM Jr conceived and supervised the study.

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Authors and Affiliations

  1. University of Pittsburgh Center for Vaccine Research, Pittsburgh, PA, USA

    M L Palma, P Duangkhae, B Douradinha, I F T Viana, S M Barratt-Boyes, E J M Nascimento & E T A Marques

  2. Laboratory of Dermatology and Immunodeficiencies, School of Medicine from University of São Paulo, São Paulo, Brazil

    M L Palma, P O Rigato, T M Oshiro & A J da Silva Duarte

  3. Fondazione Ri.MED, Palermo, Italy

    B Douradinha

  4. Center of Immunology, Institute Adolfo Lutz, Secretary of Health of São Paulo, São Paulo, Brazil,

    P O Rigato

  5. Virology and Experimental Virology Department, Aggeu Magalhães Research Center, Fiocruz, Recife, Brazil

    R Dhalia & E T A Marques

  6. Department of Infectious Diseases and Microbiology, University of Pittsburgh, Pittsburgh, PA, USA

    R B Mailliard, S M Barratt-Boyes & E T A Marques

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  1. M L Palma
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Palma, M., Duangkhae, P., Douradinha, B. et al. Development of potent class II transactivator gene delivery systems capable of inducing de novo MHC II expression in human cells, in vitro and ex vivo. Gene Ther 24, 342–352 (2017). https://doi.org/10.1038/gt.2017.25

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