Residual Cancer Lymphocytes in Patients With Chronic Lymphocytic Leukemia After Therapy Show Increased Expression of Surface Antigen CD52 Detected Using.

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Residual Cancer Lymphocytes in Patients With Chronic Lymphocytic Leukemia After Therapy Show Increased Expression of Surface Antigen CD52 Detected Using Quantitative Fluorescence Cytometry Michaela Pevna, Michael Doubek, Petr Coupek, Olga Stehlikova, Martin Klabusay Clinical Lymphoma, Myeloma and Leukemia Volume 14, Issue 5, Pages 411-418 (October 2014) DOI: 10.1016/j.clml.2014.06.006 Copyright © 2014 The Authors Terms and Conditions

Figure 1 (A) CD52 Antibody Titering on CLL Cells. 2 × 105 Leukocytes in the Total Volume of 100 μL. CLL Cells are Saturated Using 5 μL of FITC-Conjugated Anti-CD52 Monoclonal Antibody. (B) Determining the Linearity of a Flow Cytometer Amplification System23 Abbreviations: CLL = Chronic Lymphocytic Leukemia; FITC = Fluorescence Intensity of Fluorescein Isothiocyanate; MFI = Mean Fluorescence Intensity. Clinical Lymphoma, Myeloma and Leukemia 2014 14, 411-418DOI: (10.1016/j.clml.2014.06.006) Copyright © 2014 The Authors Terms and Conditions

Figure 2 Quantitatively Determined CD20 Expression in 4 Groups of Patients Based on MESF-FITC Units. There Was No Significant Difference Among the Patients Abbreviations: CLL = Chronic Lymphocytic Leukemia; FITC = Fluorescence Intensity of Fluorescein Isothiocyanate; MESF = Molecules of Equivalent Soluble Fluorochrome; SLL = Small Lymphocytic Lymphoma. Clinical Lymphoma, Myeloma and Leukemia 2014 14, 411-418DOI: (10.1016/j.clml.2014.06.006) Copyright © 2014 The Authors Terms and Conditions

Figure 3 Quantitatively Determined CD52 Expression Among 4 Groups of Patients in MESF-FITC Units. The Group of Patients in Category CR/PR Differs Significantly From the Others (P < .001) Abbreviations: CLL = Chronic Lymphocytic Leukemia; FITC = Fluorescence Intensity of Fluorescein Isothiocyanate; MESF = Molecules of Equivalent Soluble Fluorochrome; SLL = Small Lymphocytic Lymphoma. Clinical Lymphoma, Myeloma and Leukemia 2014 14, 411-418DOI: (10.1016/j.clml.2014.06.006) Copyright © 2014 The Authors Terms and Conditions

Figure 4 Quantitative Fluorescence Cytometry in a Patient With MRD-Positive Complete Remission. (A) Initial FS/SS Dot Plot Used to Gate Lymphocytes; (B) CD5+CD19+ Population Gated on Lymphocytes, 2-Color Dot Plot; (C) MFI of the Target Antigen CD52 in Histogram Based on Channel FL1-FITC, Gated on CD5+19+ Lymphocytes; (D) Standard Calibration Particles on FS/SS Dot Plot; (E) 5 Peaks of Standard Particles in Histogram Based on Channel FL1-FITC; and (F) Calibration Curve Constructed From MFI Values of Each Peak of Standard Particles. The Level of CD52 Expression in the Patient is 1,199,186 MESF-FITC Abbreviations: FITC = Fluorescence Intensity of Fluorescein Isothiocyanate; FS/SS = Forward scatter/Side scatter; lin = linear scale/channel; MESF = Molecules of Equivalent Soluble Fluorochrome; MFI = Mean Fluorescence Intensity; MRD = Minimal Residual Disease; PE = phycoerythrin. Clinical Lymphoma, Myeloma and Leukemia 2014 14, 411-418DOI: (10.1016/j.clml.2014.06.006) Copyright © 2014 The Authors Terms and Conditions

Figure 5 MESF Values Plotted Against Percentage of Residual CD5+19+ Population in Group 2 Abbreviation: MESF = Molecules of Equivalent Soluble Fluorochrome. Clinical Lymphoma, Myeloma and Leukemia 2014 14, 411-418DOI: (10.1016/j.clml.2014.06.006) Copyright © 2014 The Authors Terms and Conditions