Open Access Research article

Murine CD4+CD25- cells activated in vitro with PMA/ionomycin and anti-CD3 acquire regulatory function and ameliorate experimental colitis in vivo

Anna Majowicz12*, Sander van der Marel12, Anje A te Velde3, Sybren L Meijer4, Harald Petry1, Sander J van Deventer12 and Valerie Ferreira1

Author Affiliations

1 Research and Development, uniQure B.V., Meibergdreef 61, 1105 BA, Amsterdam, The Netherlands

2 Department of Gastroenterology & Hepatology, Leiden University Medical Center, Albinusdreef 2, 2333 ZA, Leiden, The Netherlands

3 Tytgat Institute for Liver and Intestinal Diseases, Meibergdreef 69-71, 1105 BK, Amsterdam, The Netherlands

4 Department of Pathology, Academic Medical Center, Meibergdreef 9, 1105 AZ, Amsterdam, The Netherlands

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BMC Gastroenterology 2012, 12:172  doi:10.1186/1471-230X-12-172

Published: 3 December 2012

Abstract

Background

Induced regulatory T (iTreg) lymphocytes show promise for application in the treatment of allergic, autoimmune and inflammatory disorders. iTreg cells demonstrate advantages over natural Treg (nTreg) cells in terms of increased number of starting population and greater potential to proliferate. Different activation methods to generate iTreg cells result in iTreg cells that are heterogeneous in phenotype and mechanisms of suppression. Therefore it is of interest to explore new techniques to generate iTreg cells and to determine their physiological relevance.

Methods

Using phorbol myristate acetate (PMA)/ionomycin and anti-CD3 activation of CD4+CD25- cells we generated in vitro functional CD4+CD25+ iTreg (TregPMA) cells. Functionality of the generated TregPMA cells was tested in vivo in a mouse model of inflammatory bowel disease (IBD) - CD45RB transfer colitis model.

Results

TregPMA cells expressed regulatory markers and proved to ameliorate the disease phenotype in murine CD45RB transfer colitis model. The body weight loss and disease activity scores for TregPMA treated mice were reduced when compared to diseased control group. Histological assessment of colon sections confirmed amelioration of the disease phenotype. Additionally, cytokine analysis showed decreased levels of proinflammatory colonic and plasma IL-6, colonic IL-1 β and higher levels of colonic IL-17 when compared to diseased control group.

Conclusions

This study identifies a new method to generate in vitro iTreg cells (TregPMA cells) which physiological efficacy has been demonstrated in vivo.

Keywords:
IBD; CD45RB transfer; Treg; PMA/ionomycin