AB0076 Study of physiological activity of cd4+cd25+ cells during adjuvant arthritis prior to and after introduction of cryopreserved placental cells
Authors: A. Goltsev, E. Lutsenko, M. Ostankov, N. Bondarovich
Journal: Annals of the Rheumatic Diseases (2013)

<h3>Background</h3> An important role in the development of autoimmune reactions during rheumatoid arthritis (RA) is played by CD4<sup>+</sup>CD25<sup>+</sup> T-regulatory cells (T<sub>reg</sub>) possessing a suppressive activity. The ability of placental structures to produce the activation factors of T<sub>reg</sub>-cells function – IL-2, <i>TGF</i>-β, to increase the number of cells with T<sub>reg</sub> phenotype - CD4<sup>+</sup>CD25<sup>high</sup> and CD4<sup>+</sup>/Foxp3<sup>+</sup> during co-culturing in the mixed lymphocyte culture (MLC) can be the precondition of using the placental cell suspension (PCS) to recover the function of T<sub>reg</sub>-cells <i>in vivo</i>, in particular during adjuvant arthritis (AA) [1]. Application of PCS as well as other cell therapy preparations foresees a mandatory use of cryopreservation as technological stage of the material preparing. <h3>Objectives</h3> The research aim was to substantiate the possible correction of the state of CD4<sup>+</sup>CD25<sup>+</sup> T-cells in lymphoid organs at AA with cryopreserved placental cells. <h3>Methods</h3> The research was performed in 16-18 g CBA/H mice. AA was induced with subplantar injection of Freund's complete adjuvant [2]. PCS (18-19 gestation days) was cryopreserved in cryovials (Nunc, Denmark) under 10% DMSO protection (suspension cryopreserved with dimexide or suspension cryopreserved with propane diol saccharol) by means of programmable freezer UOP-1. PCS was intravenously injected to the 7<sup>th</sup> day of AA development in a dose of 1'10<sup>6</sup> cells per mouse. The content of T-reg cells (CD4<sup>+</sup>CD25<sup>+</sup>) was examined with flow cytometer (FACS Calibur, BD) in lymph nodes and spleen of animals with AA prior to and after introduction of PCS by the method of direct immune fluorescence using monoclonal antibodies to CD4 and CD25 (BD Pharmingen TM). <h3>Results</h3> The introduction of native PCS contributed to the reduced arthritis index (AI) and recovery of the content of immune competent cells in lymphoid organs. When determining the relationship between AI and changed content of T<sub>reg</sub>-cells with CD4<sup>+</sup>CD25<sup>+</sup> phenotype in lymphoid organs of mice we have found a significant rise in the number of these cells to the 7<sup>th</sup> day of AA development in lymph nodes and to the 21<sup>st</sup> day in spleen on the background of increased joint oedema in both terms. After introduction of native PCS the dynamics of changing the content of T<sub>reg</sub>-cells in spleen and lymph nodes was one-way. The introduction of both cryopreserved suspensions rendered therapeutic effect on AA course along with this the effect after introduction of the suspension cryopreserved with dimexide was related to the regulation of T<sub>reg</sub>-cells pool in spleen and after introduction of the suspension cryopreserved with propane diol saccharol was done to their content in lymph nodes. <h3>Conclusions</h3> The prospects of using the cryopreserved PCS when treating AA are related to possible correction of the state of T<sub>reg</sub>-cells population in lymphoid organs. Implementation of therapeutic effect after PCS introduction testifies to its systemic effect when treating AA. <h3>References</h3> Goltsev A.N., Lutsenko E.D., Ostankov M.V., Bondarovich N.A. Estimation of cytokine profile in adjuvant arthritis model after cryopreserved placenta cells application // Patologiya. – 2011. – Vol.8, N2. – P. 99-101. Mischenko O.Ya., Kotvitska A.A. Pharmacological efficiency of analbene emulsion in the model of adjuvant arthritis in rats // Visnyk Farmatsii. – 2001. – N3. – P. 124-125. <h3>Disclosure of Interest</h3> None Declared