Skip to main content
SpringerLink
Log in

Administration of CD4+CD25highCD127−FoxP3+ Regulatory T Cells for Relapsing-Remitting Multiple Sclerosis: A Phase 1 Study

  • Original Research Article
  • Published:
BioDrugs Aims and scope Submit manuscript

Abstract

Background

Multiple sclerosis (MS) is an immune-mediated disease in which autoimmune T conventional (Tconv) cells break the blood–brain barrier and destroy neurons of the central nervous system. It is hypothesized that CD4+CD25highCD127−FoxP3+ T regulatory (Treg) cells may inhibit this destruction through suppressive activity exerted on Tconv cells.

Methods

We present the results of a phase 1b/2a, open-label, two-arm clinical trial in 14 patients treated with autologous Treg cells for relapsing-remitting MS. The patients received either expanded ex vivo Treg cells intravenously (intravenous [IV] group, n = 11; dose 40 × 106 Treg cells/kg of body weight) or freshly isolated Treg cells intrathecally (intrathecal [IT] group, n = 3; dose 1.0 × 106 Treg cells). Importantly, patients were not treated with any other disease-modifying drugs for at least 6 months before the recruitment and during the follow-up.

Results

No severe adverse events were observed. Self-assessed quality of life (EuroQol–5 Dimensions [EQ-5D] form) did not change and did not differ significantly between the groups. A total of 12 relapses were noted in five intravenously treated patients, who had from one to three attacks per year. Three out of ten participants who completed the trial in the IV group deteriorated more than 1 point on the Expanded Disability Status Scale (EDSS) during the follow-up. At the same time, no patients in the IT group experienced a relapse or such a deterioration in the EDSS. No significant differences were found in the Multiple Sclerosis Functional Composite (MSFC) scale in both the IV and IT groups. Magnetic resonance imaging (MRI) scans revealed a significantly lower change in the T2 lesion volume in the IT group compared to the IV group. The increase in the number of new T2 lesions during the follow-up was significant for the IV group only. There were no significant changes in the level of Treg cells or Tconv cells in the peripheral blood throughout the follow-up or between the groups. Interestingly, Treg cells in all patients consisted of two different phenotypes: peripheral Treg cells Helios(−) (≈ 20%) and thymic Treg cells Helios(+) (≈ 80%). The analysis of the cytokine pattern revealed higher levels of transforming growth factor-α and proinflammatory factors MCP3, CXCL8, and IL-1RA in the IT group compared with the IV group.

Conclusions

No serious adverse events were reported in the 14 patients with MS treated with Treg cells in this study. The results suggest that IT administration is more promising than IV administration. Because of the low number of patients recruited, the statistical results may be underpowered and further studies are necessary to reach conclusions on efficacy and safety.

Trial registration

EudraCT: 2014-004320-22; registered 18 November 2014.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Dendrou CA, Fugger L, Friese MA. Immunopathology of multiple sclerosis. Nat Rev Immunol. 2015;15:545.

    Article  CAS  PubMed  Google Scholar 

  2. Havla J, Warnke C, Derfuss T, Kappos L, Hartung HP, Hohlfeld R. Interdisciplinary risk management in the treatment of multiple sclerosis. Dtsch Arztebl Int. 2016;113:879–86.

    PubMed  PubMed Central  Google Scholar 

  3. Abbas AK, Benoist C, Bluestone JA, Campbell DJ, Ghosh S, Hori S, Jiang S, Kuchroo VK, Mathis D, Roncarolo MG, Rudensky A, Sakaguchi S, Shevach EM, Vignali DA, Ziegler SF. Regulatory T cells: recommendations to simplify the nomenclature. Nat Immunol. 2013;14:307–8.

    Article  CAS  PubMed  Google Scholar 

  4. Kleinewietfeld M, Hafler DA. Regulatory T cells in autoimmune neuroinflammation. Immunol Rev. 2014;259:231–44.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Sakaguchi S, Miyara M, Costantino CM, Hafler DA. FOXP3+ regulatory T cells in the human immune system. Nat Rev Immunol. 2010;10:490–500.

    Article  CAS  PubMed  Google Scholar 

  6. Miyara M, Gorochov G, Ehrenstein M, Musset L, Sakaguchi S, Amoura Z. Human FoxP3+ regulatory T cells in systemic autoimmune diseases. Autoimmun Rev. 2011;10:744–55.

    Article  CAS  PubMed  Google Scholar 

  7. Trzonkowski P, Szmit E, Myśliwska J, Myśliwski A. Ex vivo expansion of CD4(+)CD25(+) T regulatory cells for immunosuppressive therapy. Cytometry A. 2009;75:175–88.

    Article  PubMed  Google Scholar 

  8. Campbell DJ. Control of regulatory T cell migration, function, and homeostasis. J Immunol. 2015;195:2507–13.

    Article  CAS  PubMed  Google Scholar 

  9. Gliwiński M, Iwaszkiewicz-Grześ D, Trzonkowski P. Cell-based therapies with T regulatory cells. BioDrugs. 2017;31:335–47.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  10. Trzonkowski P, Bacchetta R, Battaglia M, Berglund D, Bohnenkamp HR, ten Brinke A, Bushell A, Cools N, Geissler EK, Gregori S, Marieke van Ham S, Hilkens C, Hutchinson JA, Lombardi G, Madrigal JA, Marek-Trzonkowska N, Martinez-Caceres EM, Roncarolo MG, Sanchez-Ramon S, Saudemont A, Sawitzki B. Hurdles in therapy with regulatory T cells. Sci Transl Med. 2015;7:304–18.

    Article  Google Scholar 

  11. Trzonkowski P, Bieniaszewska M, Juścińska J, Dobyszuk A, Krzystyniak A, Marek N, Myśliwska J, Hellmann A. First-in-man clinical results of the treatment of patients with graft versus host disease with human ex vivo expanded CD4+CD25+CD127- T regulatory cells. Clin Immunol. 2009;133:22–6.

    Article  CAS  PubMed  Google Scholar 

  12. Marek-Trzonkowska N, Myśliwiec M, Dobyszuk A, Grabowska M, Techmanska I, Juscinska J, Wujtewicz MA, Witkowski P, Mlynarski W, Balcerska A, Mysliwska J, Trzonkowski P. Administration of CD4+CD25highCD127- regulatory T cells preserves β-cell function in type 1 diabetes in children. Diabetes Care. 2012;35:1817–20.

    Article  PubMed  PubMed Central  Google Scholar 

  13. Marek-Trzonkowska N, Myśliwiec M, Iwaszkiewicz-Grześ D, Gliwiński M, Derkowska I, Żalińska M, Zieliński M, Grabowska M, Zielińska H, Piekarska K, Jaźwińska-Curyłło A, Owczuk R, Szadkowska A, Wyka K, Witkowski P, Młynarski W, Jarosz-Chobot P, Bossowski A, Siebert J, Trzonkowski P. Factors affecting long-term efficacy of T regulatory cell-based therapy in type 1 diabetes. J Transl Med. 2016;14:332.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  14. Marek-Trzonkowska N, Myśliwiec M, Dobyszuk A, Grabowska M, Derkowska I, Juścińska J, Owczuk R, Szadkowska A, Witkowski P, Młynarski W, Jarosz-Chobot P, Bossowski A, Siebert J, Trzonkowski P. Therapy of type 1 diabetes with CD4+CD25highCD127- regulatory T cells prolongs survival of pancreatic islets—results of one year follow-up. Clin Immunol. 2014;153:23–30.

    Article  CAS  PubMed  Google Scholar 

  15. Marek N, Bieniaszewska M, Krzystyniak A, Juscinska J, Mysliwska J, Witkowski P, Hellmann A, Trzonkowski P. The time is crucial for ex vivo expansion of T regulatory cells for therapy. Cell Transplant. 2011;11–12:1747–58.

    Article  Google Scholar 

  16. Duffy SS, Keating BA, Perera CJ, Lees JG, Tonkin RS, Makker PGS, Carrive P, Butovsky O, Moalem-Taylor G. Regulatory T cells and their derived cytokine, interleukin-35, reduce pain in experimental autoimmune encephalomyelitis. J Neurosci. 2011;39:2326–46.

    Article  Google Scholar 

  17. McDonald WI, Compston A, Edan G, Goodkin D, Hartung HP, Lublin FD, McFarland HF, Paty DW, Polman CH, Reingold SC, Sandberg-Wollheim M, Sibley W, Thompson A, van den Noort S, Weinshenker BY, Wolinsky JS. Recommended diagnostic criteria for multiple sclerosis: guidelines from the International Panel on the diagnosis of multiple sclerosis. Ann Neurol. 2001;50:121–7.

    Article  CAS  PubMed  Google Scholar 

  18. Polman CH, Reingold SC, Banwell B, Clanet M, Cohen JA, Filippi M, Fujihara K, Havrdova E, Hutchinson M, Kappos L, Lublin FD, Montalban X, O’Connor P, Sandberg-Wollheim M, Thompson AJ, Waubant E, Weinshenker B, Wolinsky JS. Diagnostic criteria for multiple sclerosis: 2010 revisions to the McDonald criteria. Ann Neurol. 2011;69:292–302.

    Article  PubMed  PubMed Central  Google Scholar 

  19. European Medicinal Agency (2005) Guideline on clinical trials in small populations. CHMP/EWP/83561/2005s. https://www.ema.europa.eu/en/documents/scientific-guideline/guideline-clinical-trials-small-populations_en.pdf. Accessed 14 Dec 2020

  20. Clinical Study Measures. National Multiple Sclerosis Society. https://www.nationalmssociety.org/For-Professionals/Researchers/Resources-for-Researchers/Clinical-Study-Measures. Accessed 14 Dec 2020

  21. Kurtzke JF. Rating neurologic impairment in multiple sclerosis: an expanded disability status scale (EDSS). Neurology. 1983;33:1444–52.

    Article  CAS  PubMed  Google Scholar 

  22. Fischer JS, Rudick RA, Cutter GR, Reingold SC, for the National MS Society Clinical Outcomes Assessment Task Force. The multiple sclerosis functional composite measure (MSFC): an integrated approach to MS clinical outcome assessment. Multiple Sclerosis. 1999;5:244–50.

    Article  CAS  PubMed  Google Scholar 

  23. EuroQol Group. EuroQol—a new facility for the measurement of health-related quality of life. Health Policy. 1990;16:199–208.

    Article  Google Scholar 

  24. Wattjes MP, Rovira À, Miller D, Yousry TA, Sormani MP, de Stefano MP, Tintoré M, Auger C, Tur C, Filippi M, Rocca MA, Fazekas F, Kappos L, Polman C, Barkhof F, Xavier M, MAGNIMS study group. Evidence-based guidelines: MAGNIMS consensus guidelines on the use of MRI in multiple sclerosis–establishing disease prognosis and monitoring patients. Nat Rev Neurol. 2015;11:597–606.

    CAS  PubMed  Google Scholar 

  25. Rovira À, Wattjes MP, Tintoré M, Tur C, Yousry TA, Sormani MP, De Stefano N, Filippi M, Auger C, Rocca MA, Barkhof F, Fazekas F, Kappos L, Polman C, Miller D, Montalban X, MAGNIMS study group. Evidence-based guidelines: MAGNIMS consensus guidelines on the use of MRI in multiple sclerosis-clinical implementation in the diagnostic process. Nat Rev Neurol. 2015;11:471–82.

    Article  PubMed  Google Scholar 

  26. Thornton AM, Korty PE, Tran DQ, Wohlfert EA, Murray PE, Belkaid Y, Shevach EM. Expression of helios, an ikaros transcription factor family member, differentiates thymic-derived from peripherally induced Foxp3+ T regulatory cells. J Immunol. 2010;184:3433–8.

    Article  CAS  PubMed  Google Scholar 

  27. International Multiple Sclerosis Genetics Consortium, Wellcome Trust Case Control Consortium. Genetic risk and a primary role for cell-mediated immune mechanisms in multiple sclerosis. Nature. 2011;476:214–9.

    Article  CAS  Google Scholar 

  28. Raddassi K, Kent SC, Yang J, Bourcier K, Bradshaw EM, Seyfert-Margolis V, Nepom GT, Kwok WW, Hafler DA. Increased frequencies of myelin oligodendrocyte glycoprotein/MHC class II-binding CD4 cells in patients with multiple sclerosis. J Immunol. 2011;187:1039.

    Article  CAS  PubMed  Google Scholar 

  29. Nylander A, Hafler DA. Multiple sclerosis. J Clin Investig. 2012;122:1180–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Zhang X, Koldzic DN, Izikson L, Reddy J, Nazareno RF, Sakaguchi S, Kuchroo VK, Weiner HL. IL-10 is involved in the suppression of experimental autoimmune encephalomyelitis by CD25+CD4+ regulatory T cells. Int Immunol. 2004;16:249–56.

    Article  CAS  PubMed  Google Scholar 

  31. Haas J, Hug A, Viehöver A, Fritzsching B, Falk CS, Filser A, Vetter T, Milkova L, Korporal M, Fritz B, Storch-Hagenlocher B, Krammer PH, Suri-Payer E, Wildemann B. Reduced suppressive effect of CD4+CD25high regulatory T cells on the T cell immune response against myelin oligodendrocyte glycoprotein in patients with multiple sclerosis. Eur J Immunol. 2005;35:3343–52.

    Article  CAS  PubMed  Google Scholar 

  32. Venken K, Hellings N, Hensen K, Rummens JL, Medaer R, Dhooghe MB, Dubois B, Raus J, Stinissen P. Secondary progressive in contrast to relapsing-remitting multiple sclerosis patients show a normal CD4+CD25+ regulatory T-cell function and FOXP3 expression. J Neurosci Res. 2006;83:1432–46.

    Article  CAS  PubMed  Google Scholar 

  33. Feger U, Luther C, Poeschel S, Melms A, Tolosa E, Wiendl H. Increased frequency of CD4+ CD25+ regulatory T cells in the cerebrospinal fluid but not in the blood of multiple sclerosis patients. Clin Exp Immunol. 2007;147:412–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Lee MJ, Bing SJ, Choi J, Jang M, Lee G, Lee H, Chang BS, Jee Y, Lee SJ, Cho IH. IKKβ-mediated inflammatory myeloid cell activation exacerbates experimental autoimmune encephalomyelitis by potentiating Th1/Th17 cell activation and compromising blood brain barrier. Mol Neurodegener. 2016;11:54.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  35. Hoffmann PER, Boeld TJ, Doser K, Piseshka B, Andreesen R, Edinger M. Only the CD45RA+ subpopulation of CD4+CD25high T cells gives rise to homogeneous regulatory T-cell lines upon in vitro expansion. Blood. 2006;108:4260–7.

    Article  CAS  PubMed  Google Scholar 

  36. Shevach EM, Thornton AM. tTregs, pTregs, and iTregs: similarities and differences. Immunol Rev. 2014;259:88–102.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Edinger MHP. Regulatory T cells for the prevention of graft-versus-host disease: professionals defeat amateurs. Eur J Immunol. 2009;39:2966–8.

    Article  CAS  PubMed  Google Scholar 

  38. The IFNB Multiple Sclerosis Study Group. Interferon β-1b is effective in relapsing-remitting multiple sclerosis: I. Clinical results of a multicenter, randomized, double-blind, placebo-controlled trial. Neurology. 1993;43:655–61.

    Article  Google Scholar 

  39. The IFNB Multiple Sclerosis Study Group and The University of British Columbia MS/MRI Analysis Group. Interferon β-1b in the treatment of multiple sclerosis: final outcome of the randomized controlled trial. Neurology. 1995;45:1277–85.

    Article  Google Scholar 

  40. Bornstein MB, Miller AI, Slagle S. A placebo controlled, double blinded, randomized, two-center, pilot trial of Cop 1 in chronic progressive multiple sclerosis. Neurology. 1991;41:533–9.

    Article  CAS  PubMed  Google Scholar 

  41. Bar-Or A, Gold R, Kappos L, Arnold DL, Giovannoni G, Selmaj K, O’Gorman J, Stephan M, Dawson KT. Clinical efficacy of BG-12 (dimethyl fumarate) in patients with relapsing-remitting multiple sclerosis: subgroup analyses of the DEFINE study. J Neurol. 2013;260:2297–305.

    Article  CAS  PubMed  Google Scholar 

  42. Bornstein MB, Miller A, Slagle S, Weitzman M, Crystal H, Drexler E, Keilson M, Merriam A, Wassertheil-Smoller S, Spada V. A pilot trial of Cop 1 in exacerbating-remitting multiple sclerosis. N Engl J Med. 1987;317:408–14.

    Article  CAS  PubMed  Google Scholar 

  43. Fox RJ, Miller DH, Phillips JT, Hutchinson M, Havrdova E, Kita M, Yang M, Raghupathi K, Novas M, Sweetser MT, Viglietta V, Dawson KT, CONFIRM Study Investigators. Placebo-controlled phase 3 study of oral BG-12 or glatiramer in multiple sclerosis. N Engl J Med. 2012;367:1087–97.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank to Prof. Maciej Juryńczyk from University of Oxford/Medical University of Łódź for critical review of the manuscript and Mrs. Anita Dobyszuk and Mrs. Grażyna Gniłka from Medical University of Gdańsk Medical Centre for their perfect assistance with laboratory and clinical procedures.

Author information

Author notes

  1. Kamil Chwojnicki, Dorota Iwaszkiewicz-Grześ and Anna Jankowska contributed equally to this study.

Authors and Affiliations

  1. Department of Neurology, Medical University of Gdańsk, Debinki 7, 80-210, Gdańsk, Poland

    Kamil Chwojnicki, Paweł Łowiec, Kamil Kowalczyk, Marlena Kubach, Bartosz Karaszewski & Walenty Nyka

  2. Department of Anaesthesiology and Intensive Care, Medical University of Gdańsk, Debinki 7, 80-210 Gdańsk, Poland

    Kamil Chwojnicki & Paulina Glasner

  3. Department of Medical Immunology, Medical University of Gdańsk, Debinki 7, 80-210, Gdańsk, Poland

    Dorota Iwaszkiewicz-Grześ, Maciej Zieliński, Mateusz Gliwiński, Justyna Sakowska, Julia Kulczycka, Piotr Trzonkowski & Piotr Trzonkowski

  4. Second Department of Radiology, Medical University of Gdańsk, Debinki 2, 80-210, Gdańsk, Poland

    Anna Jankowska, Małgorzata Grzywińska, Aleksandra Konarzewska & Edyta Szurowska

  5. Department of Human Physiology, Medical University of Gdańsk, Debinki 2, 80-210, Gdańsk, Poland

    Małgorzata Grzywińska

  6. Department of Ophthalmology, Medical University of Gdańsk, Debinki 2, 80-210, Gdańsk, Poland

    Paulina Glasner

  7. Regional Center of Blood Donation and Treatment, Hoene-Wrońskiego 4, 80-210, Gdańsk, Poland

    Anna Jaźwińska-Curyłło

  8. Poltreg S.A., Trzy Lipy 3 Street, 80-172, Gdańsk, Poland

    Piotr Trzonkowski

Authors
  1. Kamil Chwojnicki
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Dorota Iwaszkiewicz-Grześ
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Anna Jankowska
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Maciej Zieliński
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Paweł Łowiec
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Mateusz Gliwiński
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Małgorzata Grzywińska
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Kamil Kowalczyk
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Aleksandra Konarzewska
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Paulina Glasner
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Justyna Sakowska
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Julia Kulczycka
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Anna Jaźwińska-Curyłło
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Marlena Kubach
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. Bartosz Karaszewski
    View author publications

    You can also search for this author in PubMed Google Scholar

  16. Walenty Nyka
    View author publications

    You can also search for this author in PubMed Google Scholar

  17. Edyta Szurowska
    View author publications

    You can also search for this author in PubMed Google Scholar

  18. Piotr Trzonkowski
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Piotr Trzonkowski.

Ethics declarations

Funding

The study was supported by the National Centre for Research and Development, Poland (grant no STRATEGMED1/233368/1/NCBR/2014 to PT).

Conflict of interest

PT and KC are co-inventors of patents related to the presented content. PT is a stakeholder and board member of POLTREG venture. Medical University of Gdańsk received payment for the license to the presented content. AJ, AJC, AK, BK, DIG, ES, JK, JS, KK, MG, MGrz, MK, MZ, PG, PŁ, and WN have no conflicts of interest relevant to the contents of this article.

Ethics approval

The study was conducted according to the Declaration of Helsinki principles. The protocol has been registered in the EudraCT database under the number 2014-004320-22 and received approval from the Institutional Review Board of the Medical University of Gdańsk (no. NKBBN/414/2012 and NKBBN/414-163/2017).

Consent to participate

Written informed consent was received from all the participants at recruitment, before any medical procedure was commenced.

Consent for publication

All the patients signed written consent that the data obtained in the study could be used for scientific publications, provided the data are anonymized.

Availability of data and material

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

Code availability

Not applicable.

Author contributions

KC was the principal investigator and contributed to the study design, protocol writing, data collection, analysis, interpretation, and writing and reviewing the report. DIG was a head of the R&D team and contributed to the study design, cell preparation, data collection, analysis, and writing and reviewing the report. AJ contributed to the study design, protocol writing, MRI data collection, analysis, interpretation, and writing and reviewing the report. MZ contributed to lab assays, data collection, and interpretation. PŁ contributed to patient care, data collection, and interpretation. MG contributed to the cell preparation, data collection, analysis, and reviewing the report. MGrz contributed to MRI data collection, analysis, and interpretation. KK contributed to patient care, data collection, and interpretation. AK contributed to MRI data collection, analysis, and interpretation. PG contributed to patient care, data collection, and interpretation. AJC contributed to clinical assessment, and data collection. JS and JK contributed to the cell preparation, data collection, and analysis. MK contributed to patient care, data collection, and interpretation. BK, WN, and ES contributed to study design, data collection, and interpretation and reviewed the report. PT is the guarantor of the study, and supervised the whole process and contributed to the study design, protocol writing, cell preparation, data collection, analysis, interpretation, and writing and reviewing the report.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (PDF 1386150 KB)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Chwojnicki, K., Iwaszkiewicz-Grześ, D., Jankowska, A. et al. Administration of CD4+CD25highCD127−FoxP3+ Regulatory T Cells for Relapsing-Remitting Multiple Sclerosis: A Phase 1 Study. BioDrugs 35, 47–60 (2021). https://doi.org/10.1007/s40259-020-00462-7

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s40259-020-00462-7

Search

Navigation