Mycophenolic acid and 6-mercaptopurine both inhibit B-cell proliferation in granulomatosis with polyangiitis patients, whereas only mycophenolic acid inhibits B-cell IL-6 production

PLoS One. 2020 Jul 9;15(7):e0235743. doi: 10.1371/journal.pone.0235743. eCollection 2020.

Abstract

Granulomatosis with polyangiitis (GPA) is an autoimmune disease affecting mainly small blood vessels. B-cells are important in the GPA pathogenesis as precursors of autoantibody-producing cells but likely also contribute (auto)antibody-independently. This has been underlined by the effectiveness of B-cell-depletion (with Rituximab) in inducing and maintaining disease remission. Mycophenolate-mofetil (MMF) and azathioprine (AZA) are immunosuppressive therapies frequently used in GPA-patients. Interestingly, MMF-treated GPA-patients are more prone to relapses than AZA-treated patients, while little is known about the influence of these drugs on B-cells. We investigated whether MMF or AZA treatment (or their active compounds) alters the circulating B-cell subset distribution and has differential effects on in vitro B-cell proliferation and cytokine production in GPA-patients that might underlie the different relapse rate. Circulating B-cell subset frequencies were determined in samples from AZA-treated (n = 13), MMF-treated (n = 12), untreated GPA-patients (n = 19) and matched HCs (n = 41). To determine the ex vivo effects of the active compounds of MMF and AZA, MPA and 6-MP respectively, on B-cell proliferation and cytokine production, PBMCs of untreated GPA-patients (n = 29) and matched HCs (n = 30) were cultured for 3-days in the presence of CpG-oligodeoxynucleotides (CpG) with MPA or 6-MP. After restimulation (with phorbol myristate acetate, calcium-ionophore), cytokine-positive B-cell frequencies were measured. Finally, to assess the effect of MMF or AZA treatment on in vitro B-cell proliferation and cytokine production, PBMCs of MMF-treated (n = 18), and AZA-treated patients (n = 28) and HCs (n = 41) were cultured with CpG. The memory B-cell frequency was increased in AZA- compared to MMF-treated patients, while no other subset was different. The active compounds of MMF and AZA showed in vitro that MPA decreased B-cell proliferation in GPA-patients and HCs. B-cell proliferation in MMF- and AZA-treated patients was not different. Finally, the IL-6+ B-cell frequency was decreased by MPA compared to 6-MP. No differences in IL-10+, IL-6+ or TNFα+ B-cell proportions or proliferation were found in MMF- and AZA-treated patients. Our results indicate that MMF could be superior to AZA in inhibiting B-cell cytokine production in GPA-patients. Future studies should assess the effects of these immunosuppressive drugs on other immune cells to elucidate mechanisms underlying the potential differences in relapse rates.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Azathioprine / pharmacology
  • B-Lymphocyte Subsets / immunology
  • B-Lymphocytes / drug effects
  • B-Lymphocytes / immunology*
  • B-Lymphocytes / metabolism
  • Cell Proliferation / drug effects
  • Cytokines / metabolism
  • Female
  • Flow Cytometry
  • Granulomatosis with Polyangiitis* / drug therapy
  • Granulomatosis with Polyangiitis* / immunology
  • Humans
  • Immunosuppressive Agents / pharmacology*
  • Interleukin-6 / metabolism
  • Male
  • Mercaptopurine / pharmacology
  • Middle Aged
  • Mycophenolic Acid / pharmacology*

Substances

  • Cytokines
  • Immunosuppressive Agents
  • Interleukin-6
  • Mercaptopurine
  • Mycophenolic Acid
  • Azathioprine

Grants and funding

Research leading to these results has received funding from the Jan Kornelis de Cock foundation (https://www.decockstichting.nl/subsidies.html). JSS is supported by personal grants from the Dutch Kidney Foundation (grant no. 13OKJ39; https://www.nierstichting.nl) and the Dutch Organization for Scientific Research (Clinical Fellow grant no. 907-14-542; https://www.zonmw.nl/nl/). WHA and PH are supported by the European Union's Horizon 2020 research and innovation program project RELENT (grant no. 668036; https://ec.europa.eu/programmes/horizon2020/en). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.