Action of methotrexate and tofacitinib on directly stimulated and bystander-activated lymphocytes

Elisa Piscianz, Vanessa Candilera, Erica Valencic, Claudia Loganes, Greta Paron, Sara De Iudicibus, Giuliana Decorti, Alberto Tommasini

Research output: Contribution to journalArticle


Chronic inflammation associated with autoimmune activation is characteristic of rheumatic diseases from childhood to adulthood. In recent decades, significant improvements in the treatment of these types of disease have been achieved using disease modifying anti-rheumatic drugs (DMARDs), such as methotrexate (MTX) and, more recently, using biologic inhibitors. The recent introduction of kinase inhibitors (for example, tofacitinib; Tofa) further increases the available ARDs. However, there are patients that do not respond to any treatment strategies, for whom combination therapies are proposed. The data regarding the combined action of different drugs is lacking and the knowledge of the mechanisms of ARDs and their actions upon pathogenic lymphocytes, which are hypothesized to sustain disease, is poor. An in vitro model of inflammation was developed in the current study, in which stimulated and unstimulated lymphocytes were cultured together, but tracked separately, to investigate the action of MTX and Tofa on the two populations. By analysing lymphocyte proliferation and activation, and cytokine secretion in the culture supernatants, it was established that, due to the presence of activated cells, unstimulated cells underwent a bystander activation that was modulated by the ARDs. Additionally, varying administration schedules were demonstrated to affect lymphocytes differently in vitro, either directly or via bystander activation. Furthermore, MTX and Tofa exerted different effects; while MTX showed an antiproliferative effect, Tofa marginally effected activation, although only a slight antiproliferative action, which could be potentiated by sequential treatment with MTX. Thus, it was hypothesized that these differences may be exploited in sequential therapeutic strategies, to maximize the anti-rheumatic effect. These findings are notable and must be accounted for, as bystander-activated cells in vivo could contribute to the spread of autoimmune activation and disease progression.

Original languageEnglish
Pages (from-to)574-582
Number of pages9
JournalMolecular Medicine Reports
Issue number1
Publication statusPublished - Jul 1 2016



  • Anti-rheumatic drugs
  • Bystander effect
  • Methotrexate
  • Pathogenic lymphocytes
  • Tofacitinib

ASJC Scopus subject areas

  • Biochemistry
  • Cancer Research
  • Genetics
  • Molecular Biology
  • Molecular Medicine
  • Oncology

Cite this

Piscianz, E., Candilera, V., Valencic, E., Loganes, C., Paron, G., De Iudicibus, S., Decorti, G., & Tommasini, A. (2016). Action of methotrexate and tofacitinib on directly stimulated and bystander-activated lymphocytes. Molecular Medicine Reports, 14(1), 574-582.