Low-dose fludarabine increases rituximab cytotoxicity in B-CLL cells by triggering caspases activation in vitro.

Rituximab maintenance therapy provides a significant benefit in patients with indolent B-cell non-Hodgkin lymphoma (NHL). Based on its efficacy in improving response to chemotherapy, the anti-CD20 antibody is currently under evaluation as maintenance therapy also in patients with B-CLL. We evaluated rituximab-mediated cytotoxicity in 10 B-CLL cases pretreated in vitro with non-cytotoxic concentrations of fludarabine. This combination induced a synergic cytotoxic effect in 8 out of 10 patients at a mean level of 26.15 +/- 13.9%, compared to 8.05 +/- 5.3% cytotoxicity observed with rituximab alone. Consistent with the viability assay, we found an increased caspase-3 activity together with activation of caspase-9 in B-CLL cells sensitive to sequential non-cytotoxic fludarabine and rituximab exposure. Non-cytotoxic fludarabine concentrations may sensitize B-CLL cells to rituximab-mediated cytotoxicity via caspase-3 activation.

What is behind the presence of anti-neutrophil cytoplasmatic antibodies in chronic liver disease?

BACKGROUND: Anti-neutrophil cytoplasm antibodies (ANCA) have been detected in serum from patients affected by autoimmune CLD, and to a lesser extent, in serum of patients affected by non autoimmune CLD. The clinical significance of ANCA in these disorders is still unclear. AIMS: To explore the clinical and diagnostic significance of ANCA in CLD. MATERIALS AND METHODS: Forty-five sera from patients affected by HCV- and HBV-related CLD (group 1), 29 sera from patients affected by alcohol-related CLD (Group 2) and 33 sera from patients affected by autoimmune-related liver diseases including chronic autoimmune hepatitis (AIH), primary sclerosing cholangitis (PSC) and primary biliary cirrhosis (PBC), (Group 3) have been studied by using IIF and ELISA. RESULTS: A low correspondence between the ANCA positivity obtained by IFI and ELISA was observed. The positivity for at least one ANCA antigen was observed in 60.0% of Group 1, in 44.8% of Group 2 and in 72.7% of Group 3. The relation between the aetiology of the disease and the number of positive ANCA tests in ELISA was not significant. The relation between the MELD score and the number of ANCA positivity in patients with cirrhosis in the autoimmune-related CLD and in the viral-related CLD has been investigated. In both Groups a significant worsening of MELD at increasing number of ANCA positivity was found. Moreover in patients without cirrhosis in Group 3 an increase in the ALT and AST activity in patients with at least one ANCA positivity was observed. CONCLUSIONS: These data suggest that the finding of ANCA by ELISA is common not only in autoimmune CLD but, also in viral-related CLD. Patients with autoimmune-related CLD express more frequently a multiple antigen reactivity as compared to those with viral-related CLD. The positivity for ANCA might have a prognostic value in patients with viral-related as well as autoimmune-related cirrhosis since it is associated with worse MELD score.

Recent advances in diagnostic technologies for autoimmune diseases.

The investigation of autoimmunity provides an interest challenge in "omics" research and, particularly, proteome research, as autoimmune diseases are common disorders of unsolved etiology that occur in a wide range of manifestations, in all of which tissues and organs are attacked by the body's own immune system. Autoantibodies are a hallmark of many autoimmune diseases and the presence of autoantibodies is a distinctive and key characteristic of autoimmune diseases. Conventionally, the study of autoimmune response has always been conducted by analysing the presence and/or concentration of individual antibodies in biological fluids. New proteomic techniques allow the simultaneous identification/measurement of different autoantibodies in sera of patients suffering from autoimmune diseases. The possibility of simultaneously measuring a number of correlated analytes appears to be very interesting for analytical reasons (reduced volumes of biological samples, reagents and low costs), logistical/managerial reasons, and pathophysiological reasons (combination of markers in disease-oriented or organ-oriented profiling). In particular, we describe data collected by using high-throughput techniques such as antigen microarrays and mass spectrometry for antibody profiling. While recently collected data demonstrate satisfactory analytical sensitivity and reproducibility, some issues such as standardization and data interpretation have to be solved before the introduction of these new and promising techniques into clinical practice.

Western blot immunoassay for HSP-70 antibodies in idiopathic tinnitus: a preliminary report.

OBJECTIVES: Our preliminary study investigated the role of nonspecific immunologic tests and immunoassay for heat shock protein 70 (HSP-70) in supporting the possibility of an autoimmune inner ear process determining idiopathic tinnitus. METHODS: Thirty-six consecutive patients with idiopathic tinnitus without other otologic or autoimmune diseases and 20 healthy blood donor subjects underwent determinations of circulating immune complexes (CICs) and other nonspecific immunologic factors and immunoassay for HSP-70. RESULTS: The mean CIC values were 4.2 microg/mL in the tinnitus patients and 0.9 microg/mL in the control group (p = .012). Thirteen of the 36 tinnitus patients and none of the control group were HSP-70-positive. Ten of the 13 HSP-70-positive patients had CIC values higher than normal. In the tinnitus group, the mean CIC values were 6.9 microg/mL and 2.6 microg/mL in the HSP-70-positive and -negative subgroups, respectively (p = .024). CONCLUSIONS: It may be hypothesized that in a significant number of cases, idiopathic tinnitus could be induced by immune response to inner ear-specific HSP-70.