T cells fail to develop in the human skin-cell explants system; an inconvenient truth

<p>Abstract</p> <p>Background</p> <p>Haplo-identical hematopoietic stem cell (HSC) transplantation is very successful in eradicating haematological tumours, but the long post-transplant T-lymphopenic phase is responsible for high morbidity and mortality rates. Clark <...

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Main Authors: Vanderlocht Joris, Lumeij Stijn BJ, Tonnaer Siebe, van der Stegen Sjoukje JC, Huijskens Mirelle JAJ, Van Elssen Catharina HMJ, Meek Bob, Kirkland Mark A, Hesselink Reinout, Germeraad Wilfred TV, Bos Gerard MJ
Format: Article
Language:English
Published: BMC 2011-02-01
Series:BMC Immunology
Online Access:http://www.biomedcentral.com/1471-2172/12/17
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author Vanderlocht Joris
Lumeij Stijn BJ
Tonnaer Siebe
van der Stegen Sjoukje JC
Huijskens Mirelle JAJ
Van Elssen Catharina HMJ
Meek Bob
Kirkland Mark A
Hesselink Reinout
Germeraad Wilfred TV
Bos Gerard MJ
author_facet Vanderlocht Joris
Lumeij Stijn BJ
Tonnaer Siebe
van der Stegen Sjoukje JC
Huijskens Mirelle JAJ
Van Elssen Catharina HMJ
Meek Bob
Kirkland Mark A
Hesselink Reinout
Germeraad Wilfred TV
Bos Gerard MJ
author_sort Vanderlocht Joris
collection DOAJ
description <p>Abstract</p> <p>Background</p> <p>Haplo-identical hematopoietic stem cell (HSC) transplantation is very successful in eradicating haematological tumours, but the long post-transplant T-lymphopenic phase is responsible for high morbidity and mortality rates. Clark <it>et al. </it>have described a skin-explant system capable of producing host-tolerant donor-HSC derived T-cells. Because this T-cell production platform has the potential to replenish the T-cell levels following transplantation, we set out to validate the skin-explant system.</p> <p>Results</p> <p>Following the published procedures, while using the same commercial components, it was impossible to reproduce the skin-explant conditions required for HSC differentiation towards mature T-cells. The keratinocyte maturation procedure resulted in fragile cells with minimum expression of delta-like ligand (DLL). In most experiments the generated cells failed to adhere to carriers or were quickly outcompeted by fibroblasts. Consequently it was not possible to reproduce cell-culture conditions required for HSC differentiation into functional T-cells. Using cell-lines over-expressing DLL, we showed that the antibodies used by Clark <it>et al. </it>were unable to detect native DLL, but instead stained 7AAD<sup>+ </sup>cells. Therefore, it is unlikely that the observed T-lineage commitment from HSC is mediated by DLL expressed on keratinocytes. In addition, we did confirm expression of the Notch-ligand Jagged-1 by keratinocytes.</p> <p>Conclusions</p> <p>Currently, and unfortunately, it remains difficult to explain the development or growth of T-cells described by Clark <it>et al.</it>, but for the fate of patients suffering from lymphopenia it is essential to both reproduce and understand how these co-cultures really "work". Fortunately, alternative procedures to speed-up T-cell reconstitution are being established and validated and may become available for patients in the near future.</p>
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spelling doaj.art-d7b2d480bf014460bc263ac33b3a926d2022-12-22T00:51:26ZengBMCBMC Immunology1471-21722011-02-011211710.1186/1471-2172-12-17T cells fail to develop in the human skin-cell explants system; an inconvenient truthVanderlocht JorisLumeij Stijn BJTonnaer Siebevan der Stegen Sjoukje JCHuijskens Mirelle JAJVan Elssen Catharina HMJMeek BobKirkland Mark AHesselink ReinoutGermeraad Wilfred TVBos Gerard MJ<p>Abstract</p> <p>Background</p> <p>Haplo-identical hematopoietic stem cell (HSC) transplantation is very successful in eradicating haematological tumours, but the long post-transplant T-lymphopenic phase is responsible for high morbidity and mortality rates. Clark <it>et al. </it>have described a skin-explant system capable of producing host-tolerant donor-HSC derived T-cells. Because this T-cell production platform has the potential to replenish the T-cell levels following transplantation, we set out to validate the skin-explant system.</p> <p>Results</p> <p>Following the published procedures, while using the same commercial components, it was impossible to reproduce the skin-explant conditions required for HSC differentiation towards mature T-cells. The keratinocyte maturation procedure resulted in fragile cells with minimum expression of delta-like ligand (DLL). In most experiments the generated cells failed to adhere to carriers or were quickly outcompeted by fibroblasts. Consequently it was not possible to reproduce cell-culture conditions required for HSC differentiation into functional T-cells. Using cell-lines over-expressing DLL, we showed that the antibodies used by Clark <it>et al. </it>were unable to detect native DLL, but instead stained 7AAD<sup>+ </sup>cells. Therefore, it is unlikely that the observed T-lineage commitment from HSC is mediated by DLL expressed on keratinocytes. In addition, we did confirm expression of the Notch-ligand Jagged-1 by keratinocytes.</p> <p>Conclusions</p> <p>Currently, and unfortunately, it remains difficult to explain the development or growth of T-cells described by Clark <it>et al.</it>, but for the fate of patients suffering from lymphopenia it is essential to both reproduce and understand how these co-cultures really "work". Fortunately, alternative procedures to speed-up T-cell reconstitution are being established and validated and may become available for patients in the near future.</p>http://www.biomedcentral.com/1471-2172/12/17
spellingShingle Vanderlocht Joris
Lumeij Stijn BJ
Tonnaer Siebe
van der Stegen Sjoukje JC
Huijskens Mirelle JAJ
Van Elssen Catharina HMJ
Meek Bob
Kirkland Mark A
Hesselink Reinout
Germeraad Wilfred TV
Bos Gerard MJ
T cells fail to develop in the human skin-cell explants system; an inconvenient truth
BMC Immunology
title T cells fail to develop in the human skin-cell explants system; an inconvenient truth
title_full T cells fail to develop in the human skin-cell explants system; an inconvenient truth
title_fullStr T cells fail to develop in the human skin-cell explants system; an inconvenient truth
title_full_unstemmed T cells fail to develop in the human skin-cell explants system; an inconvenient truth
title_short T cells fail to develop in the human skin-cell explants system; an inconvenient truth
title_sort t cells fail to develop in the human skin cell explants system an inconvenient truth
url http://www.biomedcentral.com/1471-2172/12/17
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