Fate of lymphocytes after withdrawal of tofacitinib treatment
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Updated: May 4, 2026

Lymphocyte Isolation from Human Skin for Phenotypic Analysis and Ex Vivo Cell Culture
Published on: April 8, 2016
Elisa Piscianz1, Erica Valencic1, Eva Cuzzoni2
1Department of Diagnostic Medicine, Institute for Maternal and Child Health - IRCCS "Burlo Garofolo", Trieste, Italy.
Abstract:
Tofacitinib (Tofa) is an inhibitor of Janus Kinase 3, developed for the treatment of autoimmune diseases and for the prevention of transplant rejection. Due to its selective action on proliferating cells, Tofa can offer a way to block T cell activation, without toxic effects on resting cells. However, few studies have investigated the effects of Tofa on lymphocyte activation in vitro. Our aim was to study the action of Tofa on different lymphocyte subsets after in vitro stimulation and to track the behaviour of treated cells after interruption of the treatment. Peripheral blood lymphocytes were stimulated in vitro with mitogen and treated with two concentrations of Tofa. After a first period in culture, cells were washed and further incubated for an additional time. Lymphocyte subsets, activation phenotype and proliferation were assessed at the different time frames. As expected, Tofa was able to reduce the activation and proliferation of lymphocytes in the first four days of treatment. In addition the drug led to a relative decrease of Natural Killer, B cells and CD8 T cells compared to CD4 T cells. However, treated cells were still viable after the first period in culture and begun to proliferate, strikingly, in a dose dependent manner when the drug was removed from the environment by replacing the culture medium. This novel data does not necessarily predict a similar behaviour in vivo, but can warn about the clinical use of this drug when a discontinuation of treatment with Tofa is considered for any reason.
Insights
Tofacitinib (Tofa) inhibits lymphocyte activation and proliferation in vitro. Upon drug removal, Tofa-treated cells resume proliferation in a dose-dependent manner, suggesting caution during treatment discontinuation.
Area of Science:
- Immunology
- Pharmacology
Background:
- Tofacitinib (Tofa) is a Janus Kinase 3 inhibitor used for autoimmune diseases and transplant rejection.
- Tofa selectively targets proliferating cells, potentially sparing resting cells from toxicity.
- Limited in vitro studies exist on Tofa's effects on lymphocyte activation.
Purpose of the Study:
- To investigate Tofa's impact on distinct lymphocyte subsets after in vitro stimulation.
- To monitor the behavior of Tofa-treated lymphocytes following treatment interruption.
Main Methods:
- Peripheral blood lymphocytes were stimulated in vitro with a mitogen.
- Cells were treated with two concentrations of Tofacitinib.
- Lymphocyte subsets, activation phenotype, and proliferation were assessed before and after drug removal.
Main Results:
- Tofa effectively reduced lymphocyte activation and proliferation within the initial four days.
- Tofa treatment led to a relative decrease in Natural Killer, B, and CD8 T cells compared to CD4 T cells.
- Following Tofa removal, treated cells exhibited dose-dependent proliferation, indicating viability and recovery.
Conclusions:
- In vitro, Tofacitinib suppresses lymphocyte activation but allows for recovery upon drug withdrawal.
- This observed cell behavior warrants consideration for clinical Tofa use, especially regarding treatment discontinuation.

