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JMD 2005, Vol. 7, No. 5
Copyright © 2005 American Society for Investigative Pathology & Association for Molecular Pathology

Immunoglobulin Mutational Status Detected through Single-Round Amplification of Partial V_H Region Represents a Good Prognostic Marker for Clinical Outcome in Chronic Lymphocytic Leukemia

Roberto Marasca^*, Rossana Maffei^*, Monica Morselli^*, Patrizia Zucchini^*, Ilaria Castelli^*, Silvia Martinelli^*, Marcella Fontana^*, Sara Ravanetti^*, Monica Curotti^*, Giovanna Leonardi^*, Katia Cagossi, Giovanni Partesotti and Giuseppe Torelli^*

From the Department of Oncology and Hematology, * University of Modena and Reggio Emilia, Modena; and the Internal Medical Divisions of Carpi and Sassuolo Hospitals, Modena, Italy

The immunoglobulin (Ig) mutational status in B-cell chronic lymphocytic leukemia (CLL) distinguishes two subsets of patients with different prognosis. Ig status detection is commonly performed with a panel of V_H family-specific primers. Although this method detects clonal VDJ rearrangement in virtually all cases, it is technically cumbersome and therefore not widely used clinically. Here, we describe a simple and rapid method to establish the mutational status of IgV_H in CLL. The method is based on a consensus V_H FR2 primer, used in both polymerase chain reaction (PCR) and sequencing reactions. Overall, monoclonal B-cell populations were detected in 163 of 189 CLL patients (86%). The prognostic value of IgV_H mutational status was then evaluated by analyzing survival in 146 CLL cases using different V_H homology cutoffs. CLL prognostic groups were best separated by the classical 98% cutoff: median survival was 127 and 206 months in unmutated and mutated CLL cases, respectively (P = 0.0023). V_H FR2 consensus and V_H family PCR were compared in 41 cases, correctly assigning all cases by both methods. Therefore, we suggest a sequential strategy to detect immunoglobulin mutational status in CLL patients by first using the approach described in this study followed by alternative V_H family-specific PCRs for negative cases.

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