THE INCIDENCE OF PRIMARY AND SECONDARY RESISTANCE TO THE ANTI-CYTOKINE THERAPY IN PATIENTS WITH INFLAMMATORY BOWEL DISEASE.

Long-term experience of using irifliximab in patients with inflammatory bowel disease (IBD) have shown that 20-30% of patients couldn't achieve clinical improvement (primary failure) or have developed acquired drug resistance (secondary failure). The frequency of antibody formation to infliximab is to 28-61%, for adalimumab 4-12%, to certolizumab the guests to 12.3%, for etanercept to 2.5%. The ways to overcome the primary and secondary inefficiency of anti-cytokine the(apy (loss of response to therapy) at the present time are: increasing the dose, shortening the interval between infusions of genetically engineered biological drugs, additional immunosuppressive agents, as well as switching to another biologic drug. Search of primary and secondary inefficiency of anti-cytokine therapy in patients with IBD currently remains topical and requires new approaches to the solution of this problem.

[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.

[Significance of a method for determination of deamidated gliadin peptide in the diagnosis of celiac disease].

AIM: To define the value of a new enzyme immunoassay in determining the level of anti-deamidated gliadin peptide (DGP) antibodies (Abs) in the diagnosis of celiac disease. SUBJECTS AND METHODS: One hundred and twenty-four patients treated at the Department of Intestinal Pathology, Central Research Institute of Gastroenterology, were examined. Enzyme-linked immunosorbent assay (ELISA) was employed to determine Abs to tissue transglutaminase (tTG) and DGP of the IgA and IgG classes in the sera of all the patients. The diagnosis of celiac disease was verified by the histological examination of small bowel mucosa biopsy specimens. RESULTS: The examinees were divided into 3 groups: 1) 27 patients first diagnosed with celiac disease; 2) 40 patients keeping a gluten-free diet (GFD); 3) 57 patients with other gastrointestinal diseases (a comparison group). In the patients first diagnosed with celiac disease, the detection rate of elevated titers of anti-tTG and anti-DGP Abs in the IgA class was equal and constituted 92.5%; that in the IgG class was 96.2 and 55.5%, respectively. The comparison group showed an increase in the DGP levels in the IgA and IgG classes in 4 (7%) patients and a rise in tTG concentrations in the IgA and IgG classes was seen in only 2 (3.5%) patients. CONCLUSION: In the patents first diagnosed with celiac disease, the detection rate of elevated levels of anti-DGP Abs in the IgA and IgG classes is 92.5 and 96.2%, respectively, and significantly indifferent from that of IgA and IgG anti-tTG Abs. The patients keeping GFD displayed a reduction in anti-DGP Abs. The high detection rate of IgA anti-DGP Abs in the patients first diagnosed with celiac disease allows this method to be recommended for immunological diagnosis of this disease in adults.

[Peculiarities of the clinical course of ulcerative colitis, depending on the spectrum of cross-linked autoantibodies from the same family to antigens of neutrophils].

There were found that patients with ulcerative colitis (UC) in the presence of two types of autoantibodies to antigens of neutrophils is observed in less pronounced activity in the course of the disease compared with those patients with a spectrum of autoantibodies one of the wider family. The patients with three or more types of antibodies from the same family, cross-linked, ware accompanied by an increase in clinical and endoscopic disease activity. It is shown that a--greater range of autoantibodies one family determines not only the degree of activity of the YAK, but the nature and intensity of its extraintestinal manifestations. Identify different spectra of autoantibodies one family to antigens of neutrophils, cross-linked, allows us to predict the nature of the disease in each particular patient.

[Clinical immunological diagnostics of overlap syndrome during autoimmune hepatic disorders].

The complex determination of serum autoantibodies to hepatic antigens using enzyme immunoassay and immunoblot allows to increase the frequency of overlap syndrome identification during autoimmune hepatic disorders and its early diagnostics, that has a big clinical, diagnostic and prognostic importance. The levels of overlap autoantibodies combine with biochemical index and with disease activity and intensity of autoimmune processes during overlap syndrome of primary biliary cirrhosis/autoimmune hepatitis (PBC/AIH).

[Elevated levels of circulating immune complexes -C3d and-C1q as a criterion for acute extraintestinal manifestations of ulcerative colitis].

During ulcerative colitis in the exacerbation phase the high level of circulating immune complexes -C1q and -C3d (CIC-C1q and -C3d) was detected in 35.4% and 28.9% of patients respectively. This raise of CIC-C1q and -C3d level was accompanied by increase of clinical and endoscopic activity of ulcerative colitis (AC). In 45.4% of AC patients with extraintestinal onsets CIC-C3d was detected and only in 11.7% of patients CIC-C1q was found. These data confirm that activation of the humoral immunity accompanied by increase in formation of immune complexes is characteristic for AC patients with extraintestinal onsets. CIC emergence in serum of AC patients is considered as a prognostically unfavorable sign indicating that the disease may be accompanied by extraintestinal onsets.

[Serum calprotectin as a marker for determining the activity of the inflammatory process and the effectiveness of therapy in inflammatory bowel disease].

The increase in the concentration level of calprotectin in serum (ICP) in patients with inflammatory bowel disease (IBD) is closely associated with increased rates of acute phase of inflammation and combined with worsening of clinical and endoscopic disease activity. In the acute stage IBD concentration UPC hung on the degree of inflammatory activity, not localization. Test with the act is a highly sensitive method for assessing the degree of inflammatory activity and the effectiveness of therapy in IBD. After transplantation of mesenchymal stromal cells from bone marrow and standard therapy largely reduced levels SKP than selective immunosuppressive therapy with infliximab (INFL) in patients with IBD.

[Immunologic and oncological safety of autologous and allogenic mesenchymal bone marrow stromal cells].

It is indubitable that autologous transplantation of mesenchymal stromal cells (MSCs) is the golden standard for cell therapy. But also it is still in interest to explore the possibilities of allogeneic MSCs transplantation, because of their special role in lymphopoiesis, particularly in the positive selection of T-lymphocytes.

[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.

[Antibodies to Saccharomyces cerevisiae as predictors of the complicated flow of Crohn disease].

Among the 143 patients in the acute phase of IBD the ASCA detected in 34 people, accounting for 23.8%, and in 60 patients with CD the increased level of ASCA was found in 24 people (40%), in 83 patients with UC the increased level of ASCA was found in 9 (10, 8%). The appearance of antibodies to Saccharomyces cerevisiae in the blood serum of patients with CD is regarded as a prognostically unfavorable sign, indicating that the disease may have a complicated course. Also, high titers of ASCA combined with complicated CD, requiring surgical treatment.

[New possibilities for overcoming the secondary inefficiency of anti-cytokine therapy in patients with inflammatory bowel diseases].

A search for ways to overcome the secondary inefficiency of anti-cytokine therapy (ACT) with infliximab (IFX) in patients with inflammatory bowel diseases (IBD) remains relevant and determines the need for new approaches to solving this problem. The secondary inefficiency of ACT has been found to depend on the level of antibodies to IFX (anti-IFX Ab). The Department of Intestinal Pathology, Central Research Institute of Gastroenterology, is investigating the mechanisms for the occurrence of primary and secondary inefficiency of ACT, as well as ways to overcome them by cultured allogenic bone marrow mesenchymal stromal cells (MSC). In the framework of the searching investigation evaluating the efficiency and safety of MSC in patients with IBD, the investigators revealed that was a phenomenon of a decrease in anti-IFX Ab and came to the conclusion that the secondary inefficiency of ACT should be overcome in a patient with ulcerative colitis (UC). The elevated anti-IFX Ab levels were directly associated with the worsening clinical and endoscopic picture of UC and with the enhanced activity of an inflammatory process. The administration of cultures MSC contributed to lower anti-IFX Ab levels, overcome secondary inefficiency (an escape phenomenon) during ACT, and enhanced IFX sensitivity. The clinical observation indicated that MSC administration reduced anti-IFX concentrations and promoted UC remission during IFX therapy. Thus, MSC transplantation can be considered as a promising method for overcoming the secondary inefficiency of ACT, which aids in increasing the previously lost response to anti-inflammatory therapy.

[Fecal calprotectin as a biomarker effectiveness of various interventions in patients with inflammatory bowel disease].

Among the chronic diseases of the gastrointestinal tract of the special place occupied by inflammatory bowel disease (IBD), in which the lining of the intestine produces a significant number of neutrophils, which has prompted researchers and clinicians use a protein derived from neutrophils as a biomarker for the assessment of the intestinal wall and the effectiveness of treatment in patients IBD. One of these proteins is calprotectin (CP), which can be considered as a biomarker of activation, destruction and loss of neutrophil cells, to a lesser extent-- the activated monocytes and macrophages. Various studies have shown that the concentration of fecal calprotectin (FCP) correlates well with endoscopic and histological parameters of intestinal inflammation. Test the FCP can be used in healthy first-degree relatives of patients with IBD to assess the possible presence of subclinical variant of intestinal inflammation in this population. Thus, a simple test of the FCP can reduce the needs of various expensive and invasive method, including costs associated with them, especially in younger patients, where in terms of differential diagnosis of IBD is often not included neoplasia of the intestine. FCP is a non-invasive, inexpensive and at the same time, highly sensitive and specific biomarker that can be used successfully in the diagnosis, evaluation of the efficacy of treatment and predicting recurrence.

[Diagnostic value of autoantibodies to asialoglycoprotein receptor in autoimmune hepatitis].

Among 106 patients with autoimmune liver disease in 12 (11.3%) were found autoantibodies to asialoglycoprotein receptor (anti-ASGPR): in 5 (9.1%) of 55 with PBC, and 4 (17.4%) of 23 with AIH and 3 (10.7%) of 28 patients with syndrome chiasm (PBC + AIH). Detection of antibodies against the ASGPR is an effective method for diagnosis of autoimmune hepatitis. Further improvement of this method allows for monitoring and disease. Anti-ASGPR levels correlate with biochemical parameters and with the severity and manifestation autoimmune processes in patients with autoimmune hepatitis.

[Immunosuppressive effect of bone marrow MSC transplantation in patients with ulcerative colitis with six kinds of one family autoantibodies with cross-relationship between each other].

The 6 types of cross-linked autobodies of one family where identified during relapsing course of Ulcerativ Colities (UC) acompanied with deterioration of clinical and endoscopic activity and increasing rate of acut inflammatory phase (CRP, number of leukocytes and erythrocyte sedimentation rate) of the disease. On the background of transplantation of mesenchymal bone marrow stromal cells (BM MSC), and despit the identification of six types of autoantibodies to antigens of neutrophils, was observed moderate activity of UC and low concentration of autoantibodies than in immunosuppressive therapy without BM MSC transplantation. Discovered anti-inflammatory effect of BM MSCs transplantation in UC may be explained by the systemic influence of immunosuppressive effect: it is known that the BM MSCs inhibit dendritic cells, T-and B-lymphocytes participating in the immune response, activate regulatory T-cells, which produce antinflammatory cytokines, IL-10 and TGF-1beta, which suppress the inflammatory process.

[Immunological criteria for evaluation of nutritional support in ulcerative colitis].

The article presents the data on the assessment of nutritional status, taking into account humoral immunologic criteria of patients with ulcerative colitis.

[Transplantation of mesenchymal bone marrow stromal cells as new opportunity to overcome ineffectiveness of primary anti-cytokine therapy in Crohn's disease].

Clinical observations and studies have shown that among 34 patients with Crohn's disease (CD) under infliximab (INFX) therapy in 8.8% cases was observed the loss of response to anticytokine therapy (so-called primary ineffectiveness of anti-cytokine therapy (ACT)). The increasing of INFX dosage up to 10 mg/kg of body weight failed to achieve clinical and endoscopic remission. It should be noted that, despite the long anticytokine therapy in these patients the antibodies to INFX were not observed and INFX concentration in blood serum practically was not determined. In CD patients with primary ineffectiveness of anti-cytokine therapy (ACT) was detected clinical deterioration of CD: increase of WR Best Index (CDAI), appearance of pain, fever, increase of stool frequency with blood and mucus, improving performance of acute phase of inflammation. Transplantation of mesenchymal bone marrow stromal cells (MSC) enhanced the level of INFX in the serum and overcoming the primary ineffectiveness of anti-cytokine therapy (ACT) and increase in sensitivity to Infliximab.

[Calprotectin concentration in stool samples as a determinant of the activity degree of inflammatory process in inflammatory bowel diseases].

At IBD in the exacerbation phase was detected the increase of fecal calprotectin (FC) level in 98% of patients. With increasing of clinical disease activity in patients with UC as well as CD was marked a significantly increased content of calprotectin in stool samples, which was accompanied by increase of indicators of inflammation acute phase: rising RRF, leukocytosis, an increase of frequency of stool with blood and mucus, fever and abdominal pain. In the phase of exacerbation the increase in concentration of CRP depends on the degree of inflammatory activity, rather than on lesion localization. The highest concentration of CRP was revealed at a high degree of IBD activity with stool frequency up to 8-10 times/day with impurity of blood and abdominal pain. At moderate activity of IBD, it is less expressed diarrhea (stool frequency 2-3 times a day), without blood, detected lower lever of PCF concentration--from 250 to 380 ug/g. A study of calprotectin concentrations in stool samples is considered to be reliable and sensitive method for evaluation of inflammatory activity in patients with inflammatory bowel disease.

[Immune response to biological therapy of inflammatory bowel diseases].

AIM: To study biological (cell and anticytokine) therapy-induced changes in the levels of proinflammatory cytokines in patients with inflammatory bowel diseases (IBD). SUBJECTS AND METHODS: Forty-four patients with chronic continuous or chronic recurrent IBD were examined. According to the performed therapy, the patients were divided into 3 groups: 1) 16 patients who took infliximab; 2) 14 patients who received combination anti-inflammatory therapy with the cultured mesenchymal stromal cells (MSC) being administered; 3) 14 patients who had standard anti-inflammatory therapy with 5-aminosalycilic acid preparations and glucocorticosteroids. The concentrations of tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (INF-gamma), and interleukins (IL)-2, -5, -8, -12, and -15 were determined in the patients' sera before and 2 months after therapy initiation. RESULTS: The elevation in the serum levels of the proinflammatory cytokines TNF-alpha, INF-gamma, and IL-2, -5, -8, -12, and -15 indicates their implication in the pathogenesis of ulcerative colitis and Crohn's disease. Their levels may evaluate both the activity of an inflammatory process and the efficiency of the therapy. The higher level of these cytokines is accompanied by the enhanced activity of diseases, which may be used to diagnose their activity, to predict the course of IBD, and to perform adequate therapy. The decreased level of the proinflammatory cytokines is indicative of the efficiency of the therapy in patients with IBD. CONCLUSION: By reducing TNF-alpha levels, infliximab therapy results in a decrease in the concentrations of other proinflammatory cytokines (IL-1, -2, -5, -8), thus lowering the inflammatory activity of IBD. MSC transplantation also reduces the level of most proinflammatory cytokines, thus diminishing the intensity of immunopathological processes, which is shown by positive changes in the clinical picture of the disease.

[Level of adalimumab and its antibody titers define the effectiveness of the biological (anticytokine) therapy in Crohn's disease].

Formation of immune antibodies to adalimumab (ADM) in patients with Crohn's disease currently has high clinical value, as antibodies (AT) influence on bioavailability, pharmacokinetic, pharmacodynamic indicators, that eventually affects expression of clinical response to treatment. The results of this study indicate that after treatment with adalimumab in 12-14 weeks in any case there was not an increase of antibodies to adalimumab. Synchronous study of concentration of adalimumab (ADA) and antibodies to ADA in serum to determine the effectiveness of anti-cytokine therapy with adalimumab in Crohn's disease.