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. 2013 Dec;140(6):813-8.
doi: 10.1309/AJCPYFQ4XMGJD6TI.

Quantification of expression of antigens targeted by antibody-based therapy in chronic lymphocytic leukemia

Prashant R Tembhare  1 , Gerald Marti, Adrian Wiestner, Heba Degheidy, Mohammed Farooqui, Robert J Kreitman, Gregory A Jasper, Constance M Yuan, David Liewehr, David Venzon, Maryalice Stetler-Stevenson
Affiliations

Quantification of expression of antigens targeted by antibody-based therapy in chronic lymphocytic leukemia

Prashant R Tembhare et al. Am J Clin Pathol. 2013 Dec.

Abstract

Objectives: Anti-CD20 (rituximab), anti-CD52 (alemtuzumab), anti-CD22 (BL22, HA22), and anti-CD25 (Oncotac) are therapeutic options that are the mainstay or in preclinical development for the treatment of chronic lymphocytic leukemia (CLL). Studies suggest that levels of surface antigen expression may affect response to monoclonal antibody-based therapy.

Methods: Using the flow cytometric Quantibrite method (BD Biosciences, San Jose, CA) to determine antibodies bound per cell, we quantified the levels of surface expression of CD20, CD22, CD25, and CD52 in CLL cells from 28 untreated patients.

Results: The CLL cells in all cases expressed CD20, CD22, and CD52 but 4 (14%) cases were negative for CD25. Although the ranking of levels of expression from highest to lowest was CD52, CD20, CD22, and CD25, the level of antigen expression on any specific case could not be accurately predicted.

Conclusions: Quantification of antigens might be useful in evaluating new antigens to target for therapy and may provide a systematic approach to selecting individualized therapy in CLL.

Keywords: Alemtuzumab; Antibody therapy; Chronic lymphocytic leukemia; Quantitative flow cytometry; Rituximab.

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Figures

Figure 1
Figure 1. Flow Cytometric Quantification of Levels of Antigen Expression in CLL
Flow cytometric analysis of CLL cells is demonstrated in Case 2. A: B-cell analysis gate is drawn around CD19 positive B-cells. B. Data from B-cell analysis gate in A is shown. Second CLL analysis gate is drawn around CD19 positive and CD5 positive B-cells. C. x-axis PE antibody staining fluorescence, y-axis cell number. C. Histogram peaks of saturation staining with anti-CD52PE, anti-CD20PE, anti-CD22PE and anti-CD25PE on cells in CLL analysis gate labeled directly.
See this image and copyright information in PMC

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