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AB0076 Study of physiological activity of cd4+cd25+ cells during adjuvant arthritis prior to and after introduction of cryopreserved placental cells
  1. A. Goltsev1,
  2. E. Lutsenko1,
  3. M. Ostankov1,
  4. N. Bondarovich1
  1. 1Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkov, Ukraine

Abstract

Background An important role in the development of autoimmune reactions during rheumatoid arthritis (RA) is played by CD4+CD25+ T-regulatory cells (Treg) possessing a suppressive activity. The ability of placental structures to produce the activation factors of Treg-cells function – IL-2, TGF-β, to increase the number of cells with Treg phenotype - CD4+CD25high and CD4+/Foxp3+ 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 Treg-cells in vivo, 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.

Objectives The research aim was to substantiate the possible correction of the state of CD4+CD25+ T-cells in lymphoid organs at AA with cryopreserved placental cells.

Methods 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 7th day of AA development in a dose of 1’106 cells per mouse.

The content of T-reg cells (CD4+CD25+) 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).

Results 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 Treg-cells with CD4+CD25+ phenotype in lymphoid organs of mice we have found a significant rise in the number of these cells to the 7th day of AA development in lymph nodes and to the 21st 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 Treg-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 Treg-cells pool in spleen and after introduction of the suspension cryopreserved with propane diol saccharol was done to their content in lymph nodes.

Conclusions The prospects of using the cryopreserved PCS when treating AA are related to possible correction of the state of Treg-cells population in lymphoid organs. Implementation of therapeutic effect after PCS introduction testifies to its systemic effect when treating AA.

  1. 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.

  2. 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.

Disclosure of Interest None Declared

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