Mesenchymal stromal cells of bone marrow and azathioprine in Crohn's disease therapy.

AIM: To evaluate the effectiveness of MSCs therapy in patients with CD receiving azathioprine (AZA). MATERIALS AND METHODS: The study included 34 patients with inflammatory (luminal) form of CD. The 1st group of patients (n=15) received an- ti-inflammatory therapy using MSCs culture in combination with AZA. The 2nd group (n=19) received MSCs without AZA. The severity of the attack was assessed in points in accordance with the of Crohn's disease activity index (CDAI). Immunoglobulins (IgA, IgG, IgM), interleukins (IL) 1ß, 4, 10, tumor necrosis factor-α (TNF-α), interferon-γ (INF-γ), transforming growth factor-1ß (TGF-1ß), C-reactive protein (CRP), platelets and erythrocyte sedimentation rate (ESR) at 2, 6 and 12 months from the beginning of MSCs therapy. RESULTS: The initial mean CDAI in the 1st group was 337.6±17.1 points, in the 2nd group - 332.7±11.0 points (p=0.3). In both groups of pa- tients there was a significant decrease in CDAI after 2 months. From the beginning of therapy MSCs: in the 1st group to 118.9±12.4 points, in the 2nd - 120.3±14.1 points (p=0.7), after 6 months - 110.3±11.1 and 114.3±11.8 points (p=0.8), respectively. After 12 months CDAI in the 1st group was 99.9±10.8 points, in the 2nd group it was 100.6±12.1 points (p=0.8). The level of IgA, IgG, IgM was significantly lower in the group of patients with a longer history of the disease and long-term ASA. After the introduction of MSC in both groups of patients with BC, there was a tendency for the growth of pro- and anti-inflammatory cytokines, with a significantly lower level of pro-inflammatory cytokines - INF-γ, TNF-α, IL-1ß - in the 1st group, indicating potentiation of the immunosuppressive effect of MSCs and AZA, which provides a more pro- nounced anti-inflammatory effect. CONCLUSION: Transplantation of MSCs promotes an increase in the serum of patients with CD initially reduced concentration of IG, cytokines and restoring their balance as the onset of clinical remission. The combination with AZA has a more pronounced anti-inflammatory effect.

[Efficacy of adalimumab for Crohn's disease in real clinical practice]. / Effektivnost' adalimumaba pri bolezni Krona v real'noi klinicheskoi praktike.

AIM: To evaluate the efficacy and safety of adalimumab (ADA) in patients with Crohn's disease (CD) treated at the Department of Inflammatory Bowel Diseases, Moscow Clinical Research and Practical Center, and to determine the predictors of a therapy response. SUBJECTS AND METHODS: All the patients with CD treated with ADA were followed up for at least 6 months or until the drug was discontinued. Therapeutic effectiveness was evaluated at 4 weeks and 6 months after the initiation of treatment and at the end of a follow-up. Complete intestinal mucosal healing was assessed at 3 and 12 months following treatment initiation. Univariate and multivariate analyses were used to determine the predictors of treatment response. RESULTS: A clinical analysis covered 70 patients (57.1% male); the follow-up period averaged 112 weeks. Perianal fistulas were at baseline established in 22 (31.4%) patients with CD. 12 (17.4%) patients had been previously treated with infliximab (INF), 7 of them discontinued the drug for secondary loss of response and 5 for adverse reactions. 68 (97.1%) patients responded to an induction course of ADA. At 4 weeks, 6 months, and at the end of the follow-up, clinical remission occurred in 66.7, 80.4 and 67.4 % of patients with luminal CD and in 45.4, 36.5, and 36.4% of those with perianal CD, respectively. At 3 and 12 months and at the end of the follow-up, there was complete healing of the intestinal mucosa in 23.5, and 41.2 and 29.5% of cases, respectively. Six (8.8%) patients responding to the induction course needed to be optimized with ADA to 40 mg weekly. The time interval between treatment initiation and dose optimization averaged 30 weeks (range 12-120 months). There were 15 (21,4%) adverse events that were responsible for ADA discontinuation in 3 (4,2%) patients. CONCLUSION: The findings demonstrate the efficacy and safety of ADA used in clinical practice.

[Use of mesenchymal stem cells in the combination therapy of ulcerative colitis].

AIM: To compare results of treatment in patients with moderate and severe ulcerative colitis (UC), receiving standart anti-inflammatory therapy (second group) and its combination with bone marrow-derived mesenchymal stromal cells (MSC) (first group). RESULTS: Complex therapy of UC acute flare-up, including MSC did not influence on reccurence frequency, remission duration and mean value of clinical and endoscopic activity indices during 1 year of follow-up: in 1 group UC flare-up occurred in 2 (16.7%) patients, in 2 group--in 3 patients (30%). RR was 0.3 (95% CI 0/08--1.36; p=0.2; x2=1.47). After 2 years of follow-up risk of UC exacerbation in 1 group was 3 times lower than in the 2 group (p=0.03). Mean duration of remission in the 1 group was 22 months, in the 2 group--17 months (p=0.049). After 3 years of follow-up remission duration in the 1 and 2 groups were 22 and 22 months, respectively (p=0.66). Activity Index according to Rachmilevich in the 1 group was 4.75+/-1.13 points, in the 2--8.1+/-1.1 points (p=0.001). CONCLUSION: MSC application increases efficacy of anti-inflammatory treatment in patients with acute exacerbation of UC.

[Safety of mesenchymal stromal cell therapy for inflammatory bowel diseases: results of a 5-year follow-up].

AIM: To compare the safety of therapy in patients with ulcerative colitis (UC) and Crohn's disease (CD) who have received combination anti-inflammatory therapy using bone marrow mesenchymal stromal cells (MSC) and standard therapy with 5-aminosalicylic acid, glucocorticosteroids, and immunosuppressive agents. SUBJECTS AND METHODS: Unfavorable consequences were analyzed in 103 patients (56 with UC and 47 with CD) with inflammatory bowel disease (IBD) after MSC administration. The findings were compared with data obtained in 208 patients with UC and CD on standard anti-inflammatory therapy. All the patients were similar in demographic parameters, the duration of disease, the extent of intestinal injury, the nature of a course, the type and degree of disease. The analyzed groups did not include patients who had received therapy with anti-TNF-α drugs. The safety of therapy was evaluated from the presence of complications occurring during the follow-up. RESULTS: By analyzing the unfavorable consequences in 103 patients with IBD and comparing them with treatment results in 208 patients with UC and CD on standard anti-inflammatory therapy, the authors revealed no differences in the development of acute posttransfusion reactions, infectious complications, exacerbations of chronic inflammatory diseases, severe infectious complications, malignant transformation, and fatal cases in patients with UC and CD, except for those with transient fever. CONCLUSION: The results of this study demonstrate that the innovative method of cell therapy is clinically safe.

[Value of adhesion molecules for evaluating the efficiency of therapy for ulcerative colitis and Crohn's disease].

AIM: To define the value of adhesion molecules (sVCAM-1 integrin, P-selectin, E-selectin, and L-selectin) for the prediction and evaluation of the efficiency of treatment in patients with ulcerative colitis (UC) and Crohn's disease. SUBJECTS AND METHODS: Twenty-six patients with UC and 14 patients with CD were examined. Of them, 16 patients took infliximab (INF) in a dose of 5 mg/kg of body weight according to the standard scheme; 14 patients received cultured mesenchymal stem stromal cells (MSSCs) in a quantity of 150 x 10(8) cells, and 10 had azathioprine (AZA) 2 mg/kg and glucocorticosteroids (GCS) 1 mg/kg of body weight. Enzyme immunoassay was used to determine the serum concentration of the adhesion molecules (L-selectin, E-selectin, P-selectin, and sVCAM-1 integrin) before and 2 months after treatment. RESULTS: The signs of bowel inflammatory disease activity and the elevated levels of adhesion molecules whose synthesis did not occur under normal conditions remained in the patients receiving GCS and AZA. INF treatment caused a decrease in P-selectin, E-selectin, and sVCAM-1 levels to 8.9 +/- 1.0, 5.5 +/- 1.7, and 9.5 +/- 4.4 ng/ml, respectively (p < 0.001). Incorporation of MSSCs was followed by a reduction of the concentrations of P-selectin and E-selectin to 6.9 +/- 1.1 and 5.7 +/- 1.3 ng/ml, respectively (p < 0.001). The level of integrin (cVCAM-1) fell to 12.2 +/- 2.2 ng/ml (p > 0.1); that of L-selectin did not drop after MSSC administration and INF induction therapy. CONCLUSION: P-selectin, E-selectin, L-selectin, and sVCAM-1 integrin are current inflammatory markers and may be used to evaluate the efficiency of standard and biological therapies for inflammatory bowel diseases and to predict disease course.

[Cell adhesion molecules in evaluation of Crohn's disease therapy].

The treatment of inflammatory bowel diseases (IBD), which include ulcerative colitis (UC) and Crohn's disease (CD) is one of actual problems of modern gastroenterology and coloproctology. In recent years a great attention is paid to the molecules of adhesion. Adhesion proteins play a significant role in the development of inflammation in patients with IBD. They cause the migration of cells from the capillaries into the center of inflammation, i.e. do much to increase the inflammatory infiltration of the mucosa and homing of lymphocytes. Changes in the levels of adhesion factors under the influence of biological therapy have been insufficiently studied. So the aim of our study was to determine the diagnostic value of adhesion molecules--integrin-sVCAM-1 and selectins P-, E-, L- for the assessment of the effectiveness of therapy in patients with UC and CD and prognosis of the disease. 15 patients with IBD were examined (15 patients with Crohn's disease (CD)). 9 patients were treated using infliximab 5 mg/kg according to the standard scheme (0-2-6 and then every 8 weeks). 3 patients with IBD received anti-inflammatory therapy with the introduction of the culture of MSC in the number of 150 x 108 cells suspended in 200 ml of physiological solution with the addition of heparin (10 IU/ml). 3 patients received azathioprine (2 mg/kg) and glucocorticosteroids (GCS) 1 mg/kg. The clinical symptoms, the level of leukocytes, erythrocyte sedimentation rate, C-reactive protein and also were analyzed before and after the treatment with infliximab and transplantation of MSC. The status of the colonic mucosa was evaluated using colonoscopy with biopsy. The concentration of adhesion molecules L-selectin, E-selectin, P-selectin, integrin-sVCAM-1 in blood serum was analyzed using immunoenzyme method twice before the beginning of treatment and after 2 months. It is established that after the standard therapy with the use of corticosteroids and azathioprine clinical and laboratory signs of IBD activity and increased levels of adhesion molecules remained in all patients. It is reliably determined that under the influence of infliximab the levels of P-selectin, E-selectin and integrin-sVCAM-1 decrease to 8.9 +/- 1.0 ng/ml, 5.5 +/- 1.7 ng/ml, 9.5 +/- 4.4 ng/ml, respectively (p < 0.001) in all patients with IBD. This point to the suppression of the synthesis of the main inflammatory cytokine alpha-TNF. Transplantation of MSC causes significant decrease of P-selectin, E-selectin to 6.9 +/- 1.1 ng/ml and 5.7 +/- 1.3 ng/ml, respectively (p < 0.001). Integrin-sVCAM-1 has decreased slightly to 12.2 +/- 2.2 ng/ml, p > 0.1. This is associated with the onset of the maximum therapeutic effect only in 1-2 months after transplantation. The levels of P-selectin, E-selectin, integrin-sVCAM-1, reflecting the acute phase of inflammation, decreased after MSC transplantation and infliximab induction therapy. The level of L-selectin, reflecting a chronic autoimmune inflammation, practically does not decrease after the MSC transplantation (8.9 +/- 0.5 ng/ml, p < 0.05) and infliximab induction therapy (9.6 +/- 0.8 ng/ml, p > 0.1). These include the appointment of long-term infliximab therapy and repeated MSC transplantations. P-selectin, E-selectin, L-selectin, integrin-sVCAM-1 are modern markers of inflammation and may be used to assess the effectiveness of standard and biological therapy in patients with IBD, and to predict the course of the disease.

[The role of adhesion molecules for assessing the effectiveness of biological treatment of patients with inflammatory bowel disease].

One of the key links of pathological inflammatory process is the formation factors of adhesion. They play a leading role in the formation of inflammatory infiltration of the mucosa of the colon. Changes in the levels factors of adhesion under the influence of biological therapy are not well understood. In this regard, the aim of our study was to investigate the influence of biological therapy (infliximab, mesenchymal stromal cells) on the level of adhesion molecules in patients with IBD. Investigated the role of adhesion molecules to evaluate the effectiveness of treatment in this group of patients. Was examined 30 patients with IBD. Of these, 16 patients with ulcerative colitis (UC) and 14 patients with Crohn's disease (CD). Of these, 16 patients received infliximab 5 mg/kg body weight, 14 patients with IBD who underwent a comprehensive anti-inflammatory therapy with the introduction of MSC culture. Before and after treatment with infliximab, MSC transplantation was carried out a study of the clinical blood test, CRP, determined by the level of adhesion molecule L-selectin, E-selectin, P-selectin, integrin - sVCAM-1 in serum by ELISA Under the influence of infliximab for all IBD patients had significantly lower levels of P-selectin, E-selectin, integrin - sVCAM-1. In the group of patients after MSC transplantation rates of P-selectin, E-selectin was significantly decreased and the level of integrin - sVCAM-1 decreased slightly. The level of L-selectin in patients both after MSC transplantation and therapy with infliximab is practically not reduced, which serves as a reflection of chronic autoimmune inflammation, and the basis for long-term use of biological therapy in IBD. Adhesion molecule P-selectin, E-selectin, integrin - sVCAM-1 decreased more rapidly under the influence of infliximab in patients with IBD because of the mechanism of drug action (suppression of the synthesis of core inflammatory cytokine TNF-alpha). After transplantation of MSCs P-selectin, E-selectin, integrin - sVCAM-1 decreased more slowly due to the fact that the maximum positive effect of MSCs developed after 1 month. P-selectin, E-selectin, L-selectin, integrin - sVCAM-1 are the modern markers of inflammation and can be used to evaluate the effectiveness of biological therapy in IBD and the prognosis of the disease.

[Quality of life in inflammatory bowel disease patients].

OBJECTIVE: To study the quality of life (QOL) patients with inflammatory diseases (IBD), gender and age characteristics of QOL, as well as its dynamics under the influence of biological and standard therapy. MATERIAL AND METHODS: Investigation of QOL using the SF-36 was performed on 90 patients with IBD, among them 35 with Crohn's disease, 55--ulcerative colitis. Mean age 35.2 +/- 1.2 years, men--53, women--37. Severity of symptoms of IBD (by Best and Truelove): mild--7 (7.7%), moderate--69 (76.6%), severe--14 (15.7%), extraintestinal manifestations were present in 34 (37.8%) patients. After a year was conducted the survey in 31 patients (group 1) after standard therapy with 5-aminosalicylic acid (5-ASA), corticosteroids (GCS), 29 patients (Group 2)--after systemic transplantation of mesenchymal stromal cells (MSCs) of 27 patients (group 3)--Treatment with infliximab (IFX). The results. QOL was reduced in all patients in all scales of the questionnaire, including adverse changes in the psycho-emotional and social spheres. In the dynamics under the influence of the therapy, especially in achieving complete clinical and endoscopic remission, tendency to improve the performance of most QOL was more pronounced in patients with IBD treated with biological therapy MSCs and inflation, compared with patients receiving standard therapy. Identified gender differences in indicators of QOL that requires to develop psychosocial rehabilitation programs that take into account these differences. It is concluded that the need for participation of psychologists and neuropsychiatrists gastroenterologist in joint management and treatment of IBD. Was concluded the necessity of psychologists and neuropsychiatrists and gastroenterologist participation in joint management and treatment of IBD.