Increased suppressor activity of transformed ex vivo regulatory T-cells in comparison with unstimulated cells of the same donor [Povyshennaia supressornaia aktivnost' transformirovannykh ex vivo reguliatornykh T-kletok v sravnenii s nestimulirovannymi kletkami togo zhe donora]

Regulatory T-cells CD4⁺CD25⁺FoxP3⁺CD127low (Tregs) play a key role in the maintenance of tolerance to auto antigens, inhibit function of effector T and B lymphocytes, and provide a balance between effector and regulatory arms of immunity. Patients with autoimmune diseases have decreased Treg numbers and impaired suppressive activity. Transformed ex vivo autologous Tregs could restore destroyed balance of the immune system. We developed a method for Treg precursor cell cultivation. Following the method, we were able to grown up 300-400 million of Tregs cells from 50 ml of peripheral blood during a week. Transformed ex vivo Tregs are 90-95% CD4⁺CD25⁺FoxP3⁺CD127low and have increased expression of transcription genes FoxP3 and Helios. Transformed ex vivo Tregs have increased demethylation of FoxP3 promoter and activated genes of proliferation markers Cycline B1, Ki67 and LGALS 1. Transformed ex vivo Tregs have increased suppressive activity and up to 80-90% these cells secrete cytokines TNFα и IFNγ. Our data suggest transformed ex vivo autologous Tregs have genetic, immunophenotypic and functional characteristics for regulatory T-cells and further can be used for adoptive immunotherapy autoimmune diseases and inhibition of transplantation immunity.Reguliatornye T-kletki CD4⁺CD25⁺FoxP3⁺SD127low (Treg) igraiut kliuchevuiu rol' v podderzhanii tolerantnosti k autoantigenam, podavliaiut funktsiiu éffektornykh T- i V-limfotsitov i obespechivaiut balans mezhdu éffektornym i reguliatornym zvenom immuniteta. U bol'nykh s autoimmunnymi zabolevaniiami snizheno soderzhanie Treg i narushena funktsiia étikh kletok. Zamestitel'naia terapiia autologichnymi kletkami patsienta, vyrashchennymi ex vivo, mozhet vosstanovit' narushennyĭ balans immunnoĭ sistemy. Nami razrabotana metodika kul'tivirovaniia kletok-predshestvennikov Treg vne organizma cheloveka, kotoraia pozvoliaet iz 50 ml perifericheskoĭ krovi vyrastit' 300-400 mln Treg v techenie odnoĭ nedeli. V nastoiashchem issledovanii pokazano, chto po sravneniiu s Treg perifericheskoĭ krovi, vyrashchennye ex vivo kletki na 90-95% imeiut fenotip CD4⁺CD25⁺FoxP3⁺SD127low Treg i povyshennuiu ékspressiiu kliuchevykh genov transkriptsii FoxP3 i Helios. V transformirovannykh Treg povyshena stepen' demetilirovaniia v promotornom uchastke gena FoxP3 i aktivirovany geny-markery proliferatsii tsiklina V1, Ki67 i LGALS 1. Transformirovannye ex vivo Treg obladaiut povyshennoĭ supressornoĭ aktivnost'iu, i do 80-90% kletok v populiatsii sekretiruiut tsitokiny TNFα i IFNγ. Nashi dannye pokazyvaiut, chto vyrashchennye ex vivo autologichnye Treg kletki imeiut geneticheskie, markernye i funktsional'nye kharakteristiki reguliatornykh T-kletok, kotorye v budushchem mogut byt' ispol'zovany dlia adaptivnoĭ immunoterapii bol'nykh s autoimmunnymi zabolevaniiami, a takzhe dlia podavleniia transplantatsionnogo immuniteta, v chastnosti, reaktsii transplantata protiv khoziaina.