Background To study the pathogenesis, diagnosis and therapy of rheumatoid arthritis (RA) there are used numerous experimental in vivo and in vitro models. Topical issue of this problem is to study the causes of disorder and rehabilitation of immune tolerance mechanisms in RA. The concept about the role of different cells in central and peripheral tolerance formation in this pathology is often contradictory and not clear. A tolerogenic activity of products of fetoplacental complex has long been studied at the Cryopathophysiology and Immunology Department of the Institute for Problems of Cryobiology and Cryomedicine of NAS of Ukraine. The presence of a wide range of immunotropic substances in placenta is a premise to use placental cell suspension (PCS) for immunocompetent sphere recovery in autoimmune diseases.

Objectives The research aim was to determine the possibilities and features of implementation of a tolerogenic activity by native and cryopreserved placental cells in experimental models of RA development: adjuvant arthritis (AA).

Methods Research was carried out in CBA/H mice. The PCS was obtained via homogenizing the murine placenta to days 18–19 of gestation. The AA was induced by subplantar administration of the complete Freund's adjuvant. The AA development was expressed as the arthritis index. Either native (nPCS) or cryopreserved PCS were administered intravenously to day 7 after pathology induction. The PCS was cryopreserved under protection of either 10% dimethyl sulfoxide solution (suspension CD) or Propandiosakharol (suspension CP). A number of CD4⁺CD25⁺ T reg cells was determined by direct immunofluorescence using monoclonal antibodies (BD Pharmingen TM) with flow cytometry (FACS Calibur, BD) in AA animal spleen. The foxp3 gene expression in murine spleen cells was assessed by multiplex reverse transcription polymerase chain reaction (RT-PCR). In PCR we used the primer pairs to foxp3 gene and that of housekeeping: β-actin. A number of gene transcripts was compared basing on the relative semi-quantitative estimation of amplification products using Agilent Bioanalyzer 2100 (USA).

Results A disordered CD4⁺CD25⁺ cell accumulation in spleen of experimental animals with AA was established and possible use of PCS for its correction was demonstrated. The foxp3 expression level in AA animal spleen cells reduced on day 28, suggesting a contribution of this factor into AA pathogenesis within a long-term period of pathology development. During this period a decrease in joint swelling correlated with an increased content of T-reg cells and foxp3 expression level in spleen of animals after either nPCS or CD administration.

Conclusions Therapeutic effect of either native or cryopreserved PCS in AA animals is manifested on molecular level, as evidenced by an increased foxp3 expression in spleen cells after suspension administration. The effect of introduced cryopreserved placental cells as for this gene activation and, consequently, T-reg, was herewith determined by cryopreservation regimen.

References

1. Goltsev A.N., Lutsenko E.D. Ostankov M.V. Effect of different cryopreservation regimens on manifestation of immune modulating activity of placenta at development of adjuvant arthritis. Problems of Cryobiology 2008; 18(4): 456–459.

2. Haque R., Lei F. Foxp3 and Bcl-xL cooperatively promote regulatory T cell persistence and prevention of arthritis development. Arthritis Research and Therapy 2010; 12: R.66.

References

Disclosure of Interest None declared