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10.18632/oncotarget.13101

http://scihub22266oqcxt.onion/10.18632/oncotarget.13101
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suck abstract from ncbi


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pmid27821811
      Oncotarget 2016 ; 7 (48 ): 79474-79484
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  • Utilization of leukapheresis and CD4 positive selection in Treg isolation and the ex-vivo expansion for a clinical application in transplantation and autoimmune disorders #MMPMID27821811
  • Go??b K ; Grose R ; Trzonkowski P ; Wickrema A ; Tibudan M ; Marek-Trzonkowska N ; Matosz S ; Solomina J ; Ostrega D ; Michael Millis J ; Witkowski P
  • Oncotarget 2016[Nov]; 7 (48 ): 79474-79484 PMID27821811 show ga
  • Adoptive transfer of T regulatory cells (Tregs) is of great interest as a novel immunosuppressive therapy in autoimmune disorders and transplantation. Obtaining a sufficient number of stable and functional Tregs generated according to current Good Manufacturing Practice (cGMP) requirements has been a major challenge in introducing Tregs as a clinical therapy. Here, we present a protocol involving leukapheresis and CD4+ cell pre-enrichment prior to Treg sorting, which allows a sufficient number of Tregs for a clinical application to be obtained. With this method there is a decreased requirement for ex- vivo expansion. The protocol was validated in cGMP conditions. Our final Treg product passed all release criteria set for clinical applications. Moreover, during expansion Tregs presented their stable phenotype: percentage of CD4+CD25hiCD127- and CD4+FoxP3+ Tregs was > 95% and > 80%, respectively, and Tregs maintained proper immune suppressive function in vitro. Our results suggest that utilization of leukapheresis and CD4 positive selection during Treg isolation improves the likelihood of obtaining a sufficient number of high quality Treg cells during subsequent ex-vivo expansion and they can be applied clinically.
  • |Biomarkers/metabolism [MESH]
  • |CD4-Positive T-Lymphocytes/*cytology [MESH]
  • |Cell Separation [MESH]
  • |Flow Cytometry [MESH]
  • |Forkhead Transcription Factors/metabolism [MESH]
  • |Humans [MESH]
  • |Interleukin-2 Receptor alpha Subunit/metabolism [MESH]
  • |Interleukin-7 Receptor alpha Subunit/metabolism [MESH]
  • |Leukapheresis/*methods [MESH]


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