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Molecular & Cellular Proteomics 2:1331-1341, 2003.
© 2003 by The American Society for Biochemistry and Molecular Biology, Inc.

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Research

Proteomic Analysis of Chronic Lymphocytic Leukemia Subtypes with Mutated or Unmutated Ig V_H Genes^*

Duncan A. E. Cochran^,, Caroline A. Evans^,, David Blinco, John Burthem, Freda K. Stevenson^¶, Simon J. Gaskell^,|| and Anthony D. Whetton^,**

From the Leukaemia Research Fund Proteomics Facility, Department of Biomolecular Sciences, and the ^|| Michael Barber Centre for Mass Spectrometry, Department of Chemistry, University of Manchester Institute of Science and Technology, Manchester M60 1QD, and the ^¶ Molecular Immunology Laboratory, Tenovus Research Laboratory, Southampton General Hospital, Southampton SO16 6YD, United Kingdom

Chronic lymphocytic leukemia (CLL) is a common hematopoietic malignant disease with variable outcome. CLL has been divided into distinct groups based on whether somatic hypermutation has occurred in the variable region of the immunoglobulin heavy-chain locus or alternatively if the cells express higher levels of the CD38 protein. We have analyzed the proteome of 12 cases of CLL (six mutated (M-CLL) and six unmutated (UM-CLL) immunoglobulin heavy-chain loci; seven CD38-negative and five CD38-positive) using two-dimensional electrophoresis and mass spectrometry. Statistical evaluation using principal component analysis indicated significant differences in patterns of protein expression between the cases with and without somatic mutation. Specific proteins indicated by principal component analysis as varying between the prognostic groups were characterized using mass spectrometry. The levels of F-actin-capping protein ß subunit, 14-3-3 ß protein, and laminin-binding protein precursor were significantly increased in M-CLL relative to UM-CLL. In addition, primary sequence data from tandem mass spectrometry showed that nucleophosmin was present as several protein spots in M-CLL but was not detected in UM-CLL samples, suggesting that several post-translationally modified forms of nucleophosmin vary between these two sample groups. No specific differences were found between CD38-positive and -negative patient samples using the same approach. The results presented show that proteomic analysis can complement other approaches in identifying proteins that may have potential value in the biological and diagnostic distinction between important clinical subtypes of CLL.

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