Young Investigator: Dynamics of Proinflammatory Cytokines in Patients With Crohnʼs Disease Who Received Combined Therapy With Mesenchymal Stromal Cells and Azathioprine

BACKGROUND: Mesenchymal stromal cells (MSC) are used for the treatment of chronic inflammatory and autoimmune diseases in recent years, including rheumatoid arthritis (RA) and inflammatory bowel disease (IBD). In most cases, together with the MSC, patients receiving concomitant immunosuppressive therapy. It is found that immunomodulatory drugs (azathioprine (AZA), methotrexate, 6-mercaptopurine, infliximab (IPF)), regardless of the concentration, do not affect the viability, differentiation, phenotype, and ability to inhibit proliferation of MSCs peripheral blood mononuclear cells. However, studies conducted by Huang HR et al. It demonstrates that IPF rendered minimal impact on the MSC proliferation, apoptosis and cell cycle, while, azathioprine inhibited cell proliferation and induced apoptosis of MSCs in vitro. Aim: investigate dynamics of proinflammatory cytokine of patients with Crohn's disease (CD) receiving mesenchymal stromal cells (MSCs) of the bone marrow and azathioprine. METHODS: 34 patients with inflammatory (luminal) form CD were divided into two groups. The first group of patients aged 19 to 58 years old (Me-29) (n=15) was treated with anti-inflammatory therapy with MSCs culture in combination with AZA. The second group of patients with CD (n=19) aged 23 to 60 years old (Me-31) received MSCs according to the recommended scheme without AZA. To assess the effectiveness of anti-inflammatory therapy was determined by the dynamics of the level of pro-inflammatory interleukins (IL) - TNF-α, IFN-γ and IL-1β at 2, 6 and 12 months from the start of MSC therapy. Initial level of IFN-γ in the 1-st group was 110,4±12,5 pg/ml, in the 2-nd - 450,8±22,4 pg/ml (P<0,05), TNF -α in 1-st group - 13,9±1,9 pg/ml, in the 2-nd - 66,7±14,2 pg/ml (P<0,05), IL-1β in 1-st group – 5,7±0,28 pg/ml, in the 2-nd - 9,2±0,2 pg/ml (P<0,05). RESULTS: After 2 months of therapy MSCs level of IFN-γ in 1-st group was significantly decreased from baseline and was 86,5±9,1 pg/ml, the 2-nd was 96,9±12,1 pg/ml (between groups P=0,5), TNF-α in 1-st group decreased to 39,6±8,4 pg/ml, in the 2-nd - 56,5±10,7 pg/ml (between groups P=0.2), IL-1β in 1-st group -50,7±9,3 pg/ml, in the 2-nd - 56,2±10,2 pg/ml (between groups P=0,7). After 6 months of therapy MSCs level of IFN-γ in 1-st group decreased and amounted to 79,4±8,5 pg/ml, in the 2-nd - 80,8±7,3 pg/ml (between groups P=0.9), TNF -α in 1-st group decreased to 44,9±6,3 pg/ml, in the 2-nd - 49,7±10,4 pg/ml (between groups P=0,7). IL-1β in 1-st group - 45,6±7,3 pg/ml, in the 2-nd - 54,2±9,2 pg/ml (between groups P=0,45). After 12 months of initiation of therapy MSCs level of IFN-γ in 1-st group decreased and amounted to 90,8±6,5 pg/ml, in the 2-nd - 128,8±12,3 pg/ml (between groups P=0,02), TNF-α in the 1-st group - 78,9±10,5 pg/ml, in the 2-nd - 116,9±13,2 pg/ml (between groups P=0,04), IL-1β in 1-st group - 68,7±8,9 pg/ml, in the 2-nd - 96,9±9,6 pg/ml (between groups P=0,03). CONCLUSION(S): After one year of observation, combined treatment of AZA and MSC significantly reduces the level of pro-inflammatory cytokines, which may have a more pronounced anti-inflammatory therapeutic effect.