BCR-ABL fusion protein detection in peripheral blood and bone marrow samples of adult precursor B-cell acute lymphoblastic leukemia patients using the flow cytometric immunobead assay.

BACKGROUND: The ability to detect the BCR-ABL fusion gene in precursor B-cell acute lymphoblastic leukemia (pB-ALL) is essential for making accurate treatment decisions. METHODS: We used a new flow cytometric immunobead assay for BCR-ABL fusion protein detection in peripheral blood and/or bone marrow samples from 38 adult pB-ALL patients and the results were compared with polymerase chain reaction (PCR) detection of BCR-ABL transcript. RESULTS: The fusion protein was detected in peripheral blood and bone marrow samples from seven of the 38 (18%) patients, and results for both the p190 and p210 were confirmed by PCR. One case, which was positive by cytogenetics and fluorescence in situ hybridization (FISH), was negative by PCR but positive by flow cytometry. Another case, which was positive by PCR and negative by flow cytometry, was from a patient on steroid treatment. CONCLUSIONS: The cytometric immunobead assay for BCR-ABL fusion protein detection was found to be suitable for the investigation of pB-ALL patients. This assay is reliable, rapid and simple to use for peripheral blood and bone marrow samples.

Low prevalence of antibodies to cyclic citrullinated peptide in patients with inflammatory bowel disease regardless of the presence of arthritis.

OBJECTIVES: Crohn's disease (CD) and ulcerative colitis are chronic idiopathic inflammatory bowel diseases (IBD) often associated with axial and/or peripheral arthritis. Antibodies against cyclic citrullinated peptide (anti-CCP) are highly specific for rheumatoid arthritis, whereas their role in IBD remains unclear. Antitumour necrosis factor-alpha agents such as infliximab (IFX) may achieve and maintain remission of both IBD and rheumatoid arthritis; however, they may also trigger the development of various autoantibodies and autoimmune manifestations. This study aims at assessing the prevalence and clinical associations of anti-CCP antibodies in IBD patients with and without arthritis under either conventional treatment or IFX. METHODS: Eighty four consecutive patients with CD [36 (42.8%) patients on IFX scheduled maintenance therapy (5 mg/kg intravenously every 8 weeks) and 48 (57.2%) on nonbiological treatment] and 50 patients with ulcerative colitis [20 (40%) patients on IFX (as described earlier) and 30 (60%) on nonbiological treatment] were evaluated. Among these 134 patients, 48 (35.8%) patients presented with concurrent arthritis. Ninety healthy individuals matched for sex and age with the IBD patients served as controls. Anti-CCP antibodies were detected using a commercially available enzyme-linked immunosorbent assay. RESULTS: Anti-CCP antibodies were detected in one CD patient on IFX maintenance therapy without evidence of arthritis. Neither the presence of arthritis nor treatment with IFX was associated with the presence of anti-CCP antibodies in IBD patients. CONCLUSION: Our data suggest that the prevalence of anti-CCP antibodies is very low in IBD patients regardless of the presence of arthritis and/or of IFX treatment.

Distinct neutrophil subpopulations phenotype by flow cytometry in myelodysplastic syndromes.

The cardinal feature of myelodysplastic syndromes (MDS) is dysplasia involving one or more myeloid cell lineages. In the present study, we used 4-color flow cytometric analysis to investigate dysgranulopoiesis in bone marrow specimens from 65 patients with MDS. The antigen expression patterns of total neutrophil granulocytes (TNG) and of the two distinct neutrophil granulocytic subpopulations (NGSs), NGS-1 (dimmer CD45 expression) and NGS-2 (stronger CD45 expression) identified on the side scatter (SS) vs. CD45-intensity plot, were studied. The neutrophil granulocytes from patients with MDS showed characteristic antigen expression aberrancies which were more pronounced in NGS-2 subpopulation. Studying separately the NGS-2 subpopulation with the CD16/MPO/LF combination, the low CD16(+)/MPO(+) and low CD16(+)/LF(+) percentages seemed to discriminate between lower-risk and higher-risk patients with MDS in most occasions. Furthermore, a detailed assessment of the NGS-1 and NGS-2 immunophenotypic patterns revealed early dysplastic changes, not otherwise observed by standard TNG analysis, especially in cases of lower-risk MDS.

Neutrophil CD64 expression and serum IL-8: sensitive early markers of severity and outcome in sepsis.

The aim of the present study was to investigate which biomarker/s reliably assess severity and mortality early in the sepsis process. In 47 critically-ill patients within the 24h of septic onset, Interleukins (IL)-8, -1beta, -6, -10, and -12p70, tumor necrosis factor-alpha (TNF-alpha), procalcitonin (PCT) and C-reactive protein (CRP) were measured in serum. Additionally, CD64 expression was measured in neutrophils. In early sepsis, neutrophil CD64 expression and IL-8 levels are the only biomarkers that increased with sepsis severity, differentiating disease stages: sepsis, severe sepsis and septic shock (p<0.001). The biomarkers that best evaluate the severity of sepsis (via APACHE II) were CD64, IL-8 and IL-6 (p<0.01), and the severity of organ failure (via SOFA) were CD64 and IL-8 (p<0.01). CD64 expression and IL-8 levels were associated with mortality within 28-days (OR=1.3, p=0.01 for CD64 and OR=1.26, p=0.024 for IL-8 by logistic regression analysis) and ROC curve analysis showed high sensitivity and specificity for predicting sepsis stages and the 28 day mortality. We conclude that there is an early increase of neutrophil CD64 expression and IL-8 levels during sepsis. Based on this single measurement it is possible to reliably assess the stage, detect the severity and predict the 28-day mortality of sepsis.

Grape anaphylaxis: a study of 11 adult onset cases.

Reports of immunoglobulin E (IgE)-mediated allergic reactions to grapes and wine are limited in the literature. Nevertheless, grapes are widely grown and consumed in Mediterranean countries. The object of this prospective study was to present clinical features, in vivo and in vitro allergy testing, and human leukocyte antigen (HLA) serotyping in patients with recurring reactions to grapes and grape products. Eleven unrelated Greek patients, six men and five women (aged 16-44 years; mean, 26.9 years) were enrolled based on a documented history of IgE-mediated reactions to grapes, wine, or other grape products. Their evaluation included full history, reaction severity, clinical examination, skin-prick tests with food allergens and molds, serum IgE, specific IgEs to the same allergen battery, and HLA typing. Patients reported 35 grape-induced anaphylaxis episodes ranging from moderate (more than one system involved but not prominent respiratory or cardiovascular symptoms; 45.5%) to severe (serious respiratory obstruction and/or hypotension and loss of consciousness; 54.5%). A causative agent was identified: wine, 10/35 (28.6%); red grapes, 9/35 (25.7%); stuffed vine leaves, 8/35 (22.9%); raisins, 3/35 (8.6%); white grapes, 2/35 (5.7%); wine vinegar, 2/35 (5. 7%); and grape juice, 1/35 (2.9%). Other foods that induced anaphylaxis were apples (54.5%), cherries (18.6%), peaches (18.6%), and bananas (9.3%). Specific IgE values were in accordance with skin-prick tests reactivity. Concerning HLA typing, 9/11 possessed HLA-DR11(5) and -DQ7(3) and the remaining two possessed HLA-DR17(3) and -DQ2 antigens. Grapes, wine and other grape products might cause serious allergic reactions in sensitized individuals. The cosensitization and reaction incidence to other fruit allergens could be a basis for further investigation of panallergens of fruits. HLA class II antigens may contribute in genetic predisposition to these allergic reactions.