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Genetic Modification of Limbal Stem Cells to Decrease Allogeneic Immune Responses

Limbal stem cell (LSC) transplantation is the only efficient treatment for patients affected by LSC deficiency (LSCD). Allogeneic LSC transplantation is one of the most successful alternative for patients with bilateral LSCD. Nevertheless, the high variability of the human leukocyte antigens (HLA) r...

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Autores principales: Valdivia, Emilio, Bertolin, Marina, Breda, Claudia, Carvalho Oliveira, Marco, Salz, Anna Katharina, Hofmann, Nicola, Börgel, Martin, Blasczyk, Rainer, Ferrari, Stefano, Figueiredo, Constanca
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8696204/
https://www.ncbi.nlm.nih.gov/pubmed/34956181
http://dx.doi.org/10.3389/fimmu.2021.747357
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author Valdivia, Emilio
Bertolin, Marina
Breda, Claudia
Carvalho Oliveira, Marco
Salz, Anna Katharina
Hofmann, Nicola
Börgel, Martin
Blasczyk, Rainer
Ferrari, Stefano
Figueiredo, Constanca
author_facet Valdivia, Emilio
Bertolin, Marina
Breda, Claudia
Carvalho Oliveira, Marco
Salz, Anna Katharina
Hofmann, Nicola
Börgel, Martin
Blasczyk, Rainer
Ferrari, Stefano
Figueiredo, Constanca
author_sort Valdivia, Emilio
collection PubMed
description Limbal stem cell (LSC) transplantation is the only efficient treatment for patients affected by LSC deficiency (LSCD). Allogeneic LSC transplantation is one of the most successful alternative for patients with bilateral LSCD. Nevertheless, the high variability of the human leukocyte antigens (HLA) remains a relevant obstacle to long-term allogeneic graft survival. This study characterized the immunologic properties of LSCs and proposed a genetic engineering strategy to reduce the immunogenicity of LSCs and of their derivatives. Hence, LSC HLA expression was silenced using lentiviral vectors encoding for short hairpin (sh) RNAs targeting β2-microglobulin (β2M) or class II major histocompatibility complex transactivator (CIITA) to silence HLA class I and II respectively. Beside the constitutive expression of HLA class I, LSCs showed the capability to upregulate HLA class II expression under inflammatory conditions. Furthermore, LSCs demonstrated the capability to induce T-cell mediated immune responses. LSCs phenotypical and functional characteristics are not disturbed after genetic modification. However, HLA silenced LSC showed to prevent T cell activation, proliferation and cytotoxicity in comparison to fully HLA-expressing LSCs. Additionally; HLA-silenced LSCs were protected against antibody-mediated cellular-dependent cytotoxicity. Our data is a proof-of-concept of the feasibility to generate low immunogenic human LSCs without affecting their typical features. The use of low immunogenic LSCs may support for long-term survival of LSCs and their derivatives after allogeneic transplantation.
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spelling pubmed-86962042021-12-24 Genetic Modification of Limbal Stem Cells to Decrease Allogeneic Immune Responses Valdivia, Emilio Bertolin, Marina Breda, Claudia Carvalho Oliveira, Marco Salz, Anna Katharina Hofmann, Nicola Börgel, Martin Blasczyk, Rainer Ferrari, Stefano Figueiredo, Constanca Front Immunol Immunology Limbal stem cell (LSC) transplantation is the only efficient treatment for patients affected by LSC deficiency (LSCD). Allogeneic LSC transplantation is one of the most successful alternative for patients with bilateral LSCD. Nevertheless, the high variability of the human leukocyte antigens (HLA) remains a relevant obstacle to long-term allogeneic graft survival. This study characterized the immunologic properties of LSCs and proposed a genetic engineering strategy to reduce the immunogenicity of LSCs and of their derivatives. Hence, LSC HLA expression was silenced using lentiviral vectors encoding for short hairpin (sh) RNAs targeting β2-microglobulin (β2M) or class II major histocompatibility complex transactivator (CIITA) to silence HLA class I and II respectively. Beside the constitutive expression of HLA class I, LSCs showed the capability to upregulate HLA class II expression under inflammatory conditions. Furthermore, LSCs demonstrated the capability to induce T-cell mediated immune responses. LSCs phenotypical and functional characteristics are not disturbed after genetic modification. However, HLA silenced LSC showed to prevent T cell activation, proliferation and cytotoxicity in comparison to fully HLA-expressing LSCs. Additionally; HLA-silenced LSCs were protected against antibody-mediated cellular-dependent cytotoxicity. Our data is a proof-of-concept of the feasibility to generate low immunogenic human LSCs without affecting their typical features. The use of low immunogenic LSCs may support for long-term survival of LSCs and their derivatives after allogeneic transplantation. Frontiers Media S.A. 2021-12-09 /pmc/articles/PMC8696204/ /pubmed/34956181 http://dx.doi.org/10.3389/fimmu.2021.747357 Text en Copyright © 2021 Valdivia, Bertolin, Breda, Carvalho Oliveira, Salz, Hofmann, Börgel, Blasczyk, Ferrari and Figueiredo https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Immunology
Valdivia, Emilio
Bertolin, Marina
Breda, Claudia
Carvalho Oliveira, Marco
Salz, Anna Katharina
Hofmann, Nicola
Börgel, Martin
Blasczyk, Rainer
Ferrari, Stefano
Figueiredo, Constanca
Genetic Modification of Limbal Stem Cells to Decrease Allogeneic Immune Responses
title Genetic Modification of Limbal Stem Cells to Decrease Allogeneic Immune Responses
title_full Genetic Modification of Limbal Stem Cells to Decrease Allogeneic Immune Responses
title_fullStr Genetic Modification of Limbal Stem Cells to Decrease Allogeneic Immune Responses
title_full_unstemmed Genetic Modification of Limbal Stem Cells to Decrease Allogeneic Immune Responses
title_short Genetic Modification of Limbal Stem Cells to Decrease Allogeneic Immune Responses
title_sort genetic modification of limbal stem cells to decrease allogeneic immune responses
topic Immunology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8696204/
https://www.ncbi.nlm.nih.gov/pubmed/34956181
http://dx.doi.org/10.3389/fimmu.2021.747357
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