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Prepublished online as a Blood First Edition Paper on November 21, 2002; DOI 10.1182/blood-2002-06-1639.

Submitted June 4, 2002
Accepted November 13, 2002
Circulating CD20 is detectable in the plasma of patients with chronic lymphocytic leukemia and is of prognostic significance
Taghi Manshouri, Kim-Anh Do, Xuemei Wang, Francis J Giles, Susan M O'Brien, Helen Saffer, Deborah Thomas, Iman Jilani, Hagop M Kantarjian, Michael J Keating, and Maher Albitar*
Department of Hematopathology, University of Texas M. D. Anderson Cancer Center, Houston, TX, USA
Department of Biostatistics, University of Texas M. D. Anderson Cancer Center, Houston, TX, USA
Department of Leukemia, University of Texas M. D. Anderson Cancer Center, Houston, TX, USA
* Corresponding author; email: malbitar{at}mdanderson.org.
CD20 is a 33-36 KD transmembrane phosphoprotein involved in the activation, proliferation, and differentiation of B-lymphocytes. The predicted amino acid sequence of the CD20 suggests 4 transmembrane-spanning regions with both N- and C-termini located in the cytoplasm. We demonstrate herein that significant levels of circulating CD20 (cCD20) can be detected in the plasma of patients with chronic lymphocytic leukemia (CLL) and that cCD20 interferes with the binding of rituximab, a humanized anti-CD20 monoclonal antibody, to CLL cells. An enzyme-linked immunoassay (ELISA) was developed to measure circulating cCD20 levels in the plasma. We measured cCD20 levels in the plasma of 180 patients with CLL and correlated these levels with clinical characteristics and outcome. cCD20 levels correlated positively with b2-microglobulin level (P = 0.006) and percentage of CD38+ cells (P = 0.03) and negatively with platelet count (P = 0.004) and hemoglobin level (P = 0.02). Patients with advanced Rai (III/IV) or Binet (C) stage disease had significantly higher levels of cCD20 than did patients with earlier stage disease (P = 0.01, and P = 0.006, respectively). There was no correlation between cCD20 level and age, lymphocyte count, or white blood cell count. Using a recursive classification method, we found that patients with cCD20 level > 1875 nM/L had significantly shorter survival than those with cCD20 < = 1875 nM/L
(P = 0.01). The prognostic value of cCD20 was independent of Rai staging or hemoglobin level. Prospective evaluation is indicated to establish whether rituximab dosing should be adjusted according to cCD20 levels.

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