High correlation of the proteome patterns in bone marrow and peripheral blood blast cells in patients with acute myeloid leukemia

<p>Abstract</p> <p>Background</p> <p>When comparing myelogenous blasts from bone marrow and peripheral blood, immunophenotyping usually show a strong correlation of expression of surface antigens. However, it remains to be determined, whether this correlation also exists on the level of protein expression.</p> <p>Method</p> <p>Therefore, we investigated both bone marrow and peripheral blood blast cells from six patients with newly diagnosed acute myeloid leukemia (AML) using conventional two-dimensional electrophoresis in the first dimension and linear polyacrylamide gels (12%) in the second dimension. Proteins were visualized using the silver staining method and image analysis was performed using the PDQuest system.</p> <p>Results</p> <p>For each patient over 80 proteins were evaluated in the sample from peripheral blood and bone marrow. We could demonstrate that the protein expression profile of bone marrow did not significantly differ from the expression patterns of peripheral blast cells.</p> <p>Conclusion</p> <p>The proteome-set of leukemic blast cells from marrow and blood, does not differ substantially when drawn from AML patients with over 80 percent blast cells in both compartments. This indicates that in AML, blasts from peripheral blood samples can be considered suitable for investigations of the proteome using 2D-electrophoresis.</p

Allergen Micro-Bead Array for IgE Detection: A Feasibility Study Using Allergenic Molecules Tested on a Flexible Multiplex Flow Cytometric Immunoassay

Background: Allergies represent the most prevalent non infective diseases worldwide. Approaching IgE-mediated sensitizations improved much by adopting allergenic molecules instead of extracts, and by using the micro-technology for multiplex testing. Objective and Methods: To provide a proof-of-concept that a flow cytometric bead array is a feasible mean for the detection of specific IgE reactivity to allergenic molecules in a multiplex-like way. A flow cytometry Allergenic Moleculebased micro-bead Array system (ABA) was set by coupling allergenic molecules with commercially available micro-beads. Allergen specific polyclonal and monoclonal antibodies, as well as samples from 167 allergic patients, characterized by means of the ISAC microarray system, were used as means to show the feasibility of the ABA. Three hundred and thirty-six sera were tested for 1 or more of the 16 selected allergens, for a total number of 1,519 tests on each of the two systems. Results: Successful coupling was initially verified by detecting the binding of rabbit polyclonal IgG, mouse monoclonal, and pooled human IgE toward three allergens, namely nDer s 1, nPen m 1, and nPru p 3. The ABA assay showed to detect IgE t

Radioresistance of the Enhancing Effect of Cells from Carrier-Immunized Mice in an In Vitro Primary Immune Response

The in vitro primary response of mouse spleen cell suspensions to 2,4,6-trinitrophenyl(Tnp)-erythrocytes has been studied. The number of anti-Tnp plaque-forming cells that arise after antigenic stimulation in vitro is greatly enhanced by prior immunization in vivo with the carrier erythrocyte. The enhancement is antigen specific. The priming for an enhanced response can be elicited with very low antigen doses and is often apparent 24 hr after immunization. It is marked from day 3 to day 14. Spleen cells from carrier-primed mice will enhance the anti-Tnp response of normal cells when mixed cultures of the two cell populations are challenged with Tnp-erythrocytes in vitro. The carrier-primed cells mediating this enhancing effect are thymus derived. The development of the thymus-derived, carrier-specific cell population has been generally assumed to involve the antigenic stimulation of cell proliferation. It was, therefore, somewhat surprising to find that the enhancing effect of the carrier-primed cells, once they had been generated, is not inhibited by x-irradiation