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

BMC Immunology

Correspondence

Mark A Kirkland¹  Joris Vanderlocht²  Sjoukje JC van der Stegen²  Gerard MJ Bos²  Wilfred TV Germeraad²  Bob Meek²  Mirelle JAJ Huijskens²  Catharina HMJ Van Elssen³  Reinout Hesselink⁴  Siebe Tonnaer⁴  Stijn BJ Lumeij⁴ 

[1] Department of Clinical and Biomedical Sciences, Geelong, University of Melbourne, Australia;Department of Internal Medicine, Division of Hematology, Maastricht University Medical Center, Maastricht, the Netherlands;Department of Internal Medicine, Division of Hematology, Maastricht University Medical Center, Maastricht, the Netherlands;PharmaCell BV, BioPartner Building, Maastricht, the Netherlands;PharmaCell BV, BioPartner Building, Maastricht, the Netherlands;

关键词: Hematopoietic Stem Cell;    Hematopoietic Stem Cell Transplantation;    Notch Signaling;    H265 Antibody;    Hematopoetic Stem Cell;   

DOI  :  10.1186/1471-2172-12-17

 received in 2010-09-17, accepted in 2011-02-18,  发布年份 2011

来源: Springer

【 摘 要 】

BackgroundHaplo-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 et al. 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.ResultsFollowing 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 et al. were unable to detect native DLL, but instead stained 7AAD+ 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.ConclusionsCurrently, and unfortunately, it remains difficult to explain the development or growth of T-cells described by Clark et al., 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.

【 授权许可】

Unknown   
© Meek et al; licensee BioMed Central Ltd. 2011. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.