Purinergic Profiling of Regulatory T-cells in Patients With Episodic Migraine

Abstract

Objectives: Immune responses in migraine are poorly characterized, yet implicated in the disease pathogenesis. This study was carried out to characterize purinergic profiles of T-cells in patients with episodic migraine without aura (MWoA) to provide mechanistic evidence for ATP and adenosine involvement in modulation of immune regulation in migraine.Methods: Peripheral blood samples were obtained from patients with migraine (n = 16) and age-matched control subjects (n = 21). Subsets of T-cells were identified by flow cytometry based on specific membrane markers.Results: Migraine patients showed reduced total T-cell counts in the peripheral blood. Whereas the total number of CD3+CD4+, CD3+CD8+, or regulatory T lymphocytes (Treg) was not changed, the proportion of Treg CD45R0+CD62L– and CD45R0–CD62L– cells was increased. Interestingly, in migraine, less Treg cells expressed CD39 and CD73 suggesting disrupted ATP breakdown to adenosine. The negative correlations were observed between the duration of migraine and the relative number of CD73+CD39– Tregs and total number of CD73-positive CD45R0+CD62L+ Tregs.Conclusion: Obtained data indicate that T-cell populations are altered in episodic migraine and suggest the involvement of Tregs in the pathophysiology of this disorder. Reduced expression of CD39 and CD73 suggests promotion of ATP-dependent pro-inflammatory and reduction of adenosine-mediated anti-inflammatory mechanisms in migraine.

CD39⁺ EXPRESSION BY REGULATORY T CELLS IN PULMONARY SARCOIDOSIS AND LOFGREN’S SYNDROME

“…To identify major subsets among peripheral blood T cells, an algorithm earlier described by [29] was applied. In brief, at least CD45-positive 40,000 singlet events were analyzed in each sample.…”

“…In particular, naïve Tregs were defined as CD45R0 -CD62L + , central memory (CM) subset -as CD45R0 + CD62L + , effector memory (EM) -as CD45R0 + , whereas CD45RApositive terminally differentiated effector T cells (TEMRA) -as CD45R0 -CD62Lcells. Finally, these Treg subsets were analyzed for surface expression of CD39 and CD73 markers [29].…”

Profile of subpopulation composition of regulatory T lymphocytes and intestinal microbiota in patients with irritable bowel syndrome

“…In addition to the usual controlled shifts, an increase of bacteria belonging to the genera Weilonella, Lahnospiria, Streptococcus, Bifidobacteria and decrease of Bacteroides spp is present. Numerous studies of gut microbiota in IBS have noted an increase in some types of microorganisms with "pro-inflammatory" activity [24,26], including representatives of the Dorea, Ruminococcus and Clostridium genera. It should be noted, that the presence of Dorea (a member of the Lachnospiraceae family) was closely associated with an increase in pro-inflammatory cytokine production.…”

“…This set of monoclonal antibodies was used to stain 100 µl of peripheral blood in accordance with the manufacturer's recommendations. To identify the main Treg populations, as well as to analyze their expression of CD39 and CD73, an algorithm (gating strategy) was used, which was described in detail earlier [24].…”

“…In order to detect individual peripheral blood Treg subpopulations, the level of expression on the surface of cells of two antigens was evaluated: CD45R0 (short form of antigen CD45, indicating differentiation of T lymphocytes into central or effector memory cells) and CD62L (adhesion molecule allo wing T lymphocytes to enter peripheral lymphoid organs through high endothelial venules) [24]. Based on this analysis three main populations of Treg were identified: naive or thymic cells with CD45R0 -CD62L + phenotype and two populations of peripheral Treg -central and effector memory cells with CD45R0 + CD62L + and CD45R0 + CD62Lphenotypes, respectively.…”