BCL2 antibodies targeted at different epitopes detect varying levels of protein expression and correlate with frequent gene amplification in diffuse large B-cell lymphoma

Samantha L. Kendrick, Lucas Redd, Andrea Muranyi, Leigh A. Henricksen, Stacey Stanislaw, Lynette M Smith, Anamarija M. Perry, Kai Fu, Dennis D. Weisenburger, Andreas Rosenwald, German Ott, Randy D. Gascoyne, Elaine S. Jaffe, Elías Campo, Jan Delabie, Rita M. Braziel, James R. Cook, Raymond R. Tubbs, Louis M. Staudt, Wing Chung ChanChristian Steidl, Thomas M. Grogan, Lisa M. Rimsza

Research output: Contribution to journalArticle

20 Scopus citations


Patients with aggressive, BCL2 protein-positive (+) diffuse large B-cell lymphoma (DLBCL) often experience rapid disease progression that is refractory to standard therapy. However, there is potential for false-negative staining of BCL2 using the standard monoclonal mouse 124 antibody that hinders the identification of these high-risk DLBCL patients. Herein, we compare 2 alternative rabbit monoclonal antibodies (E17 and SP66) to the 124 clone in staining for BCL2 in formalin-fixed, paraffin-embedded DLBCL tissues. Overall, in 2 independent DLBCL cohorts, E17 and SP66 detected BCL2 expression more frequently than 124. In the context of MYC expression, cases identified as BCL2 (+) with SP66 demonstrated the strongest correlation with worse overall survival. The 124 clone failed to detect BCL2 expression in the majority of translocation (+), amplification (+), and activated B-cell DLBCL cases in which high levels of BCL2 protein are expected. Using dual in situ hybridization as a new tool to detect BCL2 translocation and amplification, we observed similar results as previously reported for fluorescence in situ hybridization for translocation but a higher amplification frequency, indicating that BCL2 amplification may be underreported in DLBCL. Among the discrepant cases, phosphorylation of BCL2 at T69 and/or S70 was more common than in the concordant cases and may contribute to the 124 false negatives, in addition to previously associated mutations within the epitope region. The accurate detection of BCL2 expression is important in the prognosis and treatment of DLBCL particularly with new anti-BCL2 therapies.

Original languageEnglish (US)
Pages (from-to)2144-2153
Number of pages10
JournalHuman Pathology
Issue number10
Publication statusPublished - Oct 1 2014



  • BCL2 amplification and translocation
  • BCL2 phosphorylation
  • Diffuse large B-cell lymphoma
  • Dual in situ hybridization (dual ISH)
  • E17 and SP66 BCL2 antibodies

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

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