Background and Aims

Systemic lupus erythematosus is a severe autoimmune disease caused by a combination of genetic, stochastic as well as environmental factors. A number of studies have demonstrated the influence of epigenetic mechanisms in the development of autoimmune diseases. Bregs are of particular importance for the control of autoimmune diseases – deficiency in Bregs can lead to pathological autoimmunity. As changes in the population of regulatory B-cell have been identified in patients with systemic lupus their number is a determining factor for the proper functioning of the immune system.

The aim of the study is to investigate the influence of overmethylation on the function and on the number of human regulatory B cells (Breg).

Methods

PBMCs from lupus patients were isolated and cultured in the presence of different concentrations of folic acid. The level of intracellular IL10 and the percentage of Breg were determined by flow cytometry.

Results

10 lupus patients and 10 healthy donors participated in the study. Two of the patients showed an increase in the IL10 producing Bregs after incubation with folic acid. No differences were observed in the samples of healthy volunteers.

Conclusions

A number of scientific studies confirm the involvement of epigenetic changes in the etiology of systemic lupus. Numerous scientific developments point to the role folic acid as the main modulator of gene expression. Modulation of development of systemic lupus by epigenetic pathways would be a novel scientific approach for modulation of genetic defects at molecular level.

Background and Aims

Systemic lupus erythematosus is an autoimmune syndrome characterized by the development of autoantibodies to a wide range of antigens. T cells have an important contribution in disease progression as being responsible for inflammatory cytokines secretion, B cell activation, and promoting amplification of inflammatory response. Annexin A1 is expressed by many cell types and binds to phospholipids in a Ca²⁺ dependent manner. Abnormal expression of annexin A1 was found on activated B/T cells in both murine and human autoimmunity suggesting its potential role as a therapeutic target.

The purpose of our study is to prove that it is possible to down-modulate the activity of autoreactive T and B cells by targeting them with monoclonal antibody against Annexin A1 in lupus-prone MRL/lpr mice.

Methods

Groups of lupus-prone MRL/lpr mice were treated with an anti-annexin A1 monoclonal antibody and the disease activity and survival of the animals were monitored. ELISA and ELISpot assays, RT-PCR, cell proliferation assay, flow cytometry, histological and immunofluorescence kidney analyses were used to determine the levels of cytokines, anti-dsDNA antibodies and kidney injuries.

Results

Administration of anti-annexin A1 monoclonal antibody resulted in suppression of IgG anti-dsDNA antibody production and of proteinuria, modulation of cytokine production, improved kidney histology, decreased disease activity and prolonged survival compared to the control group.

Conclusions

The anti-Annexin A1 antibody therapy targets over-activated autoreactive cells. It has a beneficial effect on earlier stage of lupus development and by using such a therapy it is possible to down-regulate the activity of lupus-associated lymphocytes.

Background and Aims

Systemic Lupus Erythematosus (SLE) is a polygenic autoimmune disorder involving multiple organs that can influence female fertility. MRL/lpr and Pristane-induced mouse models of SLE are very suitable to study female fertility compared to the healthy animals with the same background.

Methods

Lupus-like symptoms were induced through intraperitoneal injection of hydrocarbon oil pristane in non-autoimmune Balb/c mice. Flow cytometry was used for detection of activation markers and apoptosis assay. The levels of cytokines, autoantibodies and the number of autoantibody-producing plasmocytes were quantified by ELISA, ELISpot and protein array.

After hormonal ovarian stimulation, ovulated oocytes were derived from oviducts. Chromatin, tubulin and actin structures were detected by Hoechst 33258, FITC-labeled alpha-tubulin antibody and rhodamine-labeled phalloidin, respectively.

Results

A single i.p. injection of pristane leads to the development of the typical SLE symptoms such as production of different autoantibodies accompanied by massive glomerular depositions of IgG-containing immune complexes in the kidneys, and proteinuria.

The total number of obtained metaphase oocytes from lupus mice was significantly lower compared to healthy controls. The maturation rate, i.e. the proportion of eggs reaching metaphase II, was also lower for Lupus mice compared to control animals.

For each oocyte, four characteristics were described – spindle morphology, actin cap, chromosomal condensation and alignment. Many specific abnormalities in the lupus group were found.

Conclusions

Pristane-induced and MRL/lpr mouse models of lupus exhibited numerous impairments of the reproductive system which may result due to disease activity, autoantibodies or damage in molecular mechanisms through the process of reproduction.