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Genetic Immunization: A New Monoclonal Antibody for the Detection of BCL-6 Protein in Paraffin Sections

José-Francisco García, Juan-Fernando García, Lorena Maestre, Elena Lucas, Lydia Sánchez-Verde, Silvia Romero-Chala, Miguel-Ángel Piris and Giovanna Roncador

Monoclonal Antibodies Unit (J-FG,LM,EL,GR), Histology and Immunochemistry Unit (LS-V), Protein Technology Unit (SR-C), Biotechnology Program, Molecular Pathology Program (J-FG,M-AP), Spanish National Cancer Centre (CNIO), Madrid, Spain

Correspondence to: Giovanna Roncador, Monoclonal Antibodies Unit, Biotechnology Program, Centro Nacional de Investigaciones Oncológicas (Spanish National Cancer Centre), C/Melchor Fernández Almagro 3, E-28029, Madrid, Spain. E-mail: groncador{at}cnio.es

Genetic immunization can be combined with hybridoma technology to generate high-affinity monoclonal antibodies (MAbs). A new anti-BCL-6 MAb (GI191E/A8) was produced by cloning full-length BCL-6 cDNA into a eukaryotic vector and delivering this into mouse epidermis using a helium gene gun. A comparative study was made of the specificity and the effects of formalin fixation on immunohistochemistry quality of GI191E/A8 and two other anti-BCL-6 MAbs. To evaluate its possible application to differential diagnosis of lymphomas, two tissue microarrays (89 diffuse large B-cell lymphomas and 24 B-cell chronic lymphocytic leukemia cases) were stained with GI191E/A8 and another anti-BCL-6 MAb produced by conventional means. Using GI191E/A8, the detection of BCL-6 protein was significantly increased, and its specificity was independent of formalin-fixation time. Using automatic quantified analysis, the correlation between the two anti-BCL-6 MAbs tested was identical in cases with overexpression or absence of BCL-6. In cases with intermediate BCL-6 protein expression, detection with GI191E/A8 was more sensitive. A significant association of higher BCL-6 expression and longer median overall survival times in diffuse large B-cell lymphomas was found. Using conventionally produced MAbs in the same patient group, the association was not significant. (J Histochem Cytochem 54:31–38, 2006)

Key Words: BCL-6 • formalin fixed • genetic immunization • monoclonal antibody • lymphomas

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