Abstract
Acute lymphoblastic leukemia type B (B-ALL) is a neoplastic disorder that shows high mortality rates due to immature lymphocyte B-cell proliferation. B-ALL diagnosis requires identification and classification of the leukemia cells. Here, we demonstrate the use of Raman spectroscopy to discriminate normal lymphocytic B-cells from three different B-leukemia transformed cell lines (i.e., RS4;11, REH, MN60 cells) based on their biochemical features. In combination with immunofluorescence and Western blotting, we show that these Raman markers reflect the relative changes in the potential biological markers from cell surface antigens, cytoplasmic proteins, and DNA content and correlate with the lymphoblastic B-cell maturation/differentiation stages. Our study demonstrates the potential of this technique for classification of B-leukemia cells into the different differentiation/maturation stages, as well as for the identification of key biochemical changes under chemotherapeutic treatments. Finally, preliminary results from clinical samples indicate high consistency of, and potential applications for, this Raman spectroscopy approach.
Highlights
CD38 at the plasma membrane; (ii) ‘common B-ALL’, when the cells originate from late pro–B lymphoblasts or intermediate B-cell precursors, as identified by the expression of CD19, CD38, CD10, and CD79a at the plasma membrane; and (iii) ‘pre–B-ALL’, when the cells originate from more committed progenitors defined as pre–B lymphoblasts that express CD19, CD38, CD10, CD79a, CD20, CD22, and immunoglobulins at the plasma membrane[7]
The third cell model was the MN60 cell line, which represents a more differentiated B-ALL cell type, as it is classified as the L3-blast (i.e., B-cell leukemia) subtype[47,48]
Using the well-defined quadruple CD38/CD19/CD20/CD10 staining for leukemia cells[49,50], we first examined these cells under confocal fluorescence microscopy and by Western blotting (Fig. 2a,b)
Summary
CD38 at the plasma membrane; (ii) ‘common B-ALL’, when the cells originate from late pro–B lymphoblasts or intermediate B-cell precursors, as identified by the expression of CD19, CD38, CD10, and CD79a at the plasma membrane; and (iii) ‘pre–B-ALL’, when the cells originate from more committed progenitors defined as pre–B lymphoblasts that express CD19, CD38, CD10, CD79a, CD20, CD22, and immunoglobulins at the plasma membrane[7]. In the last 10 to 15 years, photonic techniques have emerged as powerful tools for determination of the invasiveness of cancer tissues during surgery[12] and for the study of the responses of biosystems at the single-cell level[13]. These methods are non-invasive[14], and they offer single-molecule detection sensitivity[15,16]. We applied the RS approach to discriminate between normal B-lymphocytes and B-lymphocyte-enriched fractions from patients with B-ALL
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