Anti-inflammatory effect of biological treatment in patients with inflammatory bowel diseases: calprotectin and IL-6 changes do not correspond to sRAGE changes.

AIM: The main objective was to examine the relationship between the soluble receptor for advanced glycation end products (sRAGE) and calprotectin concentrations in faeces and serum of patients with inflammatory bowel diseases (IBD) during biological treatment with infliximab. MATERIALS AND METHODS: A total of 29 IBD patients treated with infliximab were evaluated. Calprotectin and sRAGE in serum and faeces and serum IL-6 and CRP were measured during the induction regimen of infliximab treatment at weeks (W) 0, 2 and 10. RESULTS: At W0, a significant increase in faecal calprotectin was found in IBD compared to healthy persons (690 +/- 696 microg/g and 23 +/- 7 microg/g, respectively, p < 0.001). No clear difference was found in serum sRAGE levels in IBD cohort compared to healthy controls (772 +/- 274 pg/mL and 720 +/- 107 pg/mL, respectively, p = 0.159); however, a significant negative correlation was found between faecal calprotectin levels and serum concentrations of sRAGE in the active IBD cohort (r = -0.518, p = 0.004). In the stool eluates, sRAGE levels were non-measurable. In the group of responders-to-treatment, the initial surge in both faecal and serum calprotectin levels as well as CRP and IL-6 was followed by a significant decrease on W10. Surprisingly, no significant changeovers were seen in serum sRAGE concentrations in responders neither in W2 nor in W10. CONCLUSIONS: Unlike other examined local and systemic inflammatory markers, serum sRAGE did not change during the infliximab treatment, despite the initial correlation with the degree of mucosal inflammation.

The influence of metals on the expression of surface antigens on human lymphocytes in vitro.

OBJECTIVES: Metals have different effects on the immune functions. Through the experimental in vitro model, we studied the changes in the activation and co-stimulatory surface markers in human lymphocytes cultivated with selected metal salts. METHODS: Whole human blood was cultivated with cadmium (Cd) or zinc (Zn) sulfate for 18 hours. The number of lymphocytes positive for activation and co-stimulatory markers was evaluated by flow cytometry. RESULTS: Elevation of the CD69 and CD23 markers as well as higher expression of CD28 was found in cultures of lymphocytes incubated with Cd. In cultures incubated with Zn, minor elevation of the HLA-DR antigen expression was observed in comparison to Cd-treated cell cultures. Decrease of CD3 expression was observed after cultivation with both Cd and Zn salts. CONCLUSION: Cd and Zn exhibit different effects on the expression of human surface activation antigens and co-stimulatory molecules. Cd in non-toxic concentrations stimulated expression of early activation molecules and therefore could change the early phase of immune response. This was not the case for Zn, where the results were similar to untreated cell cultures.

Production of interleukins 10 and 12 by activated peripheral blood monocytes/macrophages in patients suffering from chronic hepatitis C virus infection with respect to the response to interferon and ribavirin treatment.

Circulating monocytes/macrophages are important for the initiation of immune responses to hepatitis C virus (HCV). Their presentation capacities and production of immunoregulatory cytokines enable them to activate cellular immune responses which is critical in determining the outcome of infection. We used flow cytometry to examine the expression of a CD80 costimulatory molecule on the surface of peripheral blood CD14+ monocytes/macrophages and to analyse the production of IL10 and IL12 by these cells. Forty-three individuals (6 asymptomatic HCV carriers, 37 patients with chronic hepatitis C (CHC)) were enrolled in this study. Thirty-seven patients with CHC (23 responders and 14 non-responders, NR) received combination (interferon+ribavirin) treatment for 52 weeks. The baseline percentage of CD14+CD80+ peripheral blood monocytes/macrophages was high in patients with CHC (P<0.001) and returned to normal after the treatment. All patients with CHC showed significantly high production of IL10 (P<0.001). In asymptomatic HCV carriers production level of this cytokine tended to be higher than in patients with CHC (P<0.001). A baseline production of IL12 was higher in asymptomatic HCV carriers and patients with CHC compared to healthy controls (P<0.001). The level of IL12 production was increased in treatment responders whereas in NR returned to normal value. Our data argue against functional impairment of circulating monocytes/macrophages during HCV infection. Furthermore, the positive therapeutic outcome following combination treatment might associate with increased production of IL12 by these cells.

Immunological predictors of different responses to combination therapy with interferon alpha and ribavirin in patients with chronic hepatitis C.

BACKGROUND: This study aimed to investigate peripheral blood CD4+ T-helper (Th) and CD8+ cytotoxic T-lymphocyte (CTL) responses to combination treatment with interferon (IFN) alpha and ribavirin in 59 patients with chronic hepatitis C, and to correlate the results with the therapy outcome. METHODS: The expression of activation molecules on the surface of CD8+ T cells and cytokine production by in-vitro activated CTLs and Th lymphocytes were examined before and at the end of the therapy, using flow cytometry. RESULTS: There were 36 complete responders to the treatment and 23 transient responders who relapsed after withdrawal of the therapy. A significant increase in the production of Th1-type cytokines [IFNgamma, interleukin 2 (IL2), and tumor necrosis factor-alpha (TNFalpha)] was found at the end of the treatment in complete responders compared with baseline values (P < 0.001). In contrast, transient responders had a marked decrease in the percentage of activated CD8+ T cells expressing CD28 or HLA-DR costimulatory molecules in peripheral blood, and a lower production of TNFalpha by CTLs and Th cells at the end of the therapy with respect to pretreatment values (P < 0.001). CONCLUSIONS: The efficacy of IFNalpha and ribavirin combination therapy for chronic hepatitis C is associated with a vigorous response of peripheral blood Th1 cells, whereas weak CTL responses at the end of the therapy might predict a further relapse of the disease.

Interference of CD95 expression on human lymphocytes.

The study presents the exogenous influence of cadmium in comparison with zinc on the apoptosis of human lymphocytes by CD95 expression and its kinetic changes. The salts of both metals were used in final concentrations of 20 microM in cell cultures with whole blood. The duration of cultivation was 18 and 90 hours. The expression of surface antigens was evaluated by flow cytometry with monoclonal antibodies. In cultures of not stimulated cells we found in average 51.54% CD95 positive lymphocytes. The kinetic study of untreated cells showed elevation after 18 hours of cultivation and a very low expression after 90 hours. The CD95 expression on lymphocytes in cell culture with cadmium and zinc was lower after 18 hours of cultivation than in untreated cells. After 90 hours cultivation we found low levels of CD95 expression on cells treated with cadmium and a great individual variability in the number of positive cells upon the influence of zinc.

Impaired deoxyribonuclease I activity in patients with inflammatory bowel diseases.

Background and Aims. Deoxyribonuclease I (DNaseI) is an endonuclease that facilitates chromatin breakdown and promotes susceptibility to autoimmune disorders. The aim of current study was to investigate serum DNase I activity in patients with inflammatory bowel diseases (IBD). Patients and Methods. A cohort of 110 IBD patients was evaluated, aged 35 ± 12 years, 77 with Crohn's disease (CD) and 33 with ulcerative colitis (UC). 50 SLE patients and 50 healthy blood donors were examined as control groups. Results. DNase I activity in IBD patients was significantly lower than in healthy individuals, but higher than in SLE patients (P < .0001). Patients with UC showed higher DNase I activity than CD patients, P = .21. DNase I activity in female patients with IBD was significantly lower than in males, P = .024; however, no differences in DNase I activity were found in relation to gender in healthy individuals. DNase I activity has shown a strong negative correlation with the serum concentration of anti-nucleosomal antibodies in the autoimmune (SLE + IBD) cohort, as well as in the separate IBD cohort. Conclusions. Reduced serum DNase I activity probably has pathogenetic consequences in IBD. Induction of autoantibodies towards nucleosomes could be a reflection of impaired DNase I activity.

Drogas citotóxicas e inmunosupresión inducida por hormonas / Citotoxic drugs and hormone-induced inmunosuppresion

Los compuestos inmunosupresores, sustancias capaces de suprimir o reducir el desarrollo de por lo menos un tipo de reacción inmune, se han empleado como quimioterapéuticos. Se probaron algunas substancias para el tratamiento inmunosupresor no específico, las cuales se pueden dividir en antimetabolitos (antagonistas del metabolismo de las purinas, azatioprina y 6-mercaptopurina, análogos del ácido fólico, metotrexate); agentes alquilantes (ciclofosfamida, clorambucil); antibióticos (ciclosporina A, actinomicina D); hormonas (glucocortidoides), y se concluyó que las cualidades inmunosupresoras de los agentes mencionados difieren en cierto grado del efecto sobre la inmunidad celular o humoral, y que la dependencia de la cronicidad de aplicación y posología del fármaco en presencia de in antígeno es una de las propiedades más importantes de los agentes inmunosupresores