Utilization of leukapheresis and CD4 positive selection in Treg isolation and the ex-vivo expansion for a clinical application in transplantation and autoimmune disorders

Karolina Golab, Randall Grose, Piotr Trzonkowski, Amittha Wickrema, Martin Tibudan, Natalia Marek-Trzonkowska, Sabrina Matosz, Julia Solomina, Diane Ostrega, J. Michael Millis, Piotr Witkowski

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

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.

LanguageEnglish
Pages79474-79484
Number of pages11
JournalOncotarget
Volume7
Issue number48
DOIs
Publication statusPublished - 1 Jan 2016

Keywords

  • CD4 cells
  • Clinical application
  • Immunosupressive therapy
  • Leukapheresis
  • Regulatory T cells

ASJC Scopus subject areas

  • Oncology

Cite this

Golab, Karolina ; Grose, Randall ; Trzonkowski, Piotr ; Wickrema, Amittha ; Tibudan, Martin ; Marek-Trzonkowska, Natalia ; Matosz, Sabrina ; Solomina, Julia ; Ostrega, Diane ; Millis, J. Michael ; Witkowski, Piotr. / Utilization of leukapheresis and CD4 positive selection in Treg isolation and the ex-vivo expansion for a clinical application in transplantation and autoimmune disorders. In: Oncotarget. 2016 ; Vol. 7, No. 48. pp. 79474-79484.
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Golab, K, Grose, R, Trzonkowski, P, Wickrema, A, Tibudan, M, Marek-Trzonkowska, N, Matosz, S, Solomina, J, Ostrega, D, Millis, JM & Witkowski, P 2016, 'Utilization of leukapheresis and CD4 positive selection in Treg isolation and the ex-vivo expansion for a clinical application in transplantation and autoimmune disorders', Oncotarget, vol. 7, no. 48, pp. 79474-79484. https://doi.org/10.18632/oncotarget.13101

Utilization of leukapheresis and CD4 positive selection in Treg isolation and the ex-vivo expansion for a clinical application in transplantation and autoimmune disorders. / Golab, Karolina; Grose, Randall; Trzonkowski, Piotr; Wickrema, Amittha; Tibudan, Martin; Marek-Trzonkowska, Natalia; Matosz, Sabrina; Solomina, Julia; Ostrega, Diane; Millis, J. Michael; Witkowski, Piotr.

In: Oncotarget, Vol. 7, No. 48, 01.01.2016, p. 79474-79484.

Research output: Contribution to journalArticle

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