@article {8355, title = {Action of methotrexate and tofacitinib on directly stimulated and bystander-activated lymphocytes.}, journal = {Mol Med Rep}, volume = {14}, year = {2016}, month = {2016 Jul}, pages = {574-82}, abstract = {

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.

}, issn = {1791-3004}, doi = {10.3892/mmr.2016.5263}, author = {Piscianz, Elisa and Candilera, Vanessa and Valencic, Erica and Loganes, Claudia and Paron, Greta and De Iudicibus, Sara and Decorti, Giuliana and Tommasini, Alberto} } @article {1798, title = {Association between BclI polymorphism in the NR3C1 gene and in vitro individual variations in lymphocyte responses to methylprednisolone.}, journal = {Br J Clin Pharmacol}, volume = {73}, year = {2012}, month = {2012 Apr}, pages = {651-5}, abstract = {

WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT: In vitro lymphocyte steroid sensitivity has been suggested as a useful tool to predict in vivo response to glucocorticoid treatment in different inflammatory chronic diseases. A correlation between genetic polymorphisms and clinical response to glucocorticoids has been demonstrated in these patients.

WHAT THIS STUDY ADDS: The BclI polymorphism in the glucocorticoid receptor (NR3C1) gene is associated with higher methylprednisolone potency in vitro. The combined evaluation of the in vitro sensitivity to methylprednisolone and BclI polymorphism could represent an aid for physicians to adjust therapy a priori. AIM To evaluate the association between the in vitro sensitivity of peripheral blood mononuclear cells (PBMCs) to methylprednisolone (MP) and the presence of genetic polymorphisms involved in glucocorticoid (GC) response.

METHODS: In vitro MP inhibition of the proliferation of lymphocytes stimulated with concanavalin A was determined. Non linear regression of dose-response data was performed computing the MP concentration required to reduce proliferation to 50\% (IC(50) ). The maximum inhibition achievable at the highest MP concentration (I(max) ) was also calculated. Moreover, the Taqman technique was used to analyze the BclI polymorphism in the NR3C1 gene and the Leu155His polymorphism in the NALP1 gene.

RESULTS: A significant association between the BclI mutated genotype and an increased in vitro sensitivity to GCs was observed.

CONCLUSIONS: The a priori evaluation of the BclI polymorphism, associated with a lymphocyte proliferation assay, could represent a useful diagnostic tool for the optimization of steroid treatment.

}, keywords = {Adaptor Proteins, Signal Transducing, Adolescent, Adult, Apoptosis Regulatory Proteins, Cyclin D1, Dose-Response Relationship, Drug, Female, Genotype, Glucocorticoids, Humans, Lymphocytes, Male, Methylprednisolone, Middle Aged, Polymorphism, Single Nucleotide, Receptors, Glucocorticoid, Young Adult}, issn = {1365-2125}, doi = {10.1111/j.1365-2125.2011.04130.x}, author = {Cuzzoni, Eva and De Iudicibus, Sara and Bartoli, Fiora and Ventura, Alessandro and Decorti, Giuliana} }