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JHC exPRESS: First Published September 7, 2005. doi:10.1369/jhc.5A6646.2005
Copyright © Histochemical Society, Inc.

A more recent version of this article appeared on January 1, 2006.
Originally published as JHC exPRESS on July 26, 2005.
doi:10.1369/jhc.5A6646.2005
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Genetic Immunization: A New Monoclonal Antibody for the Detection of BCL-6 Protein in Paraffin Sections

José-Francisco García 1, Juan-Fernando García 1, Lorena Maestre 1, Elena Lucas 1, Lydia Sánchez-Verde 1, Silvia Romero-Chala 1, Miguel-Ángel Piris 1 and Giovanna Roncador 1*

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

* To whom correspondence should be addressed. E-mail: groncador{at}cnio.es.

Submitted on February 7, 2005
Accepted on 11 July 2005


  Abstract
Genetic immunization can be combined with hybridoma technology to generate high-affinity monoclonal antibodies. A new anti-BCL-6 monoclonal antibody (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 monoclonal antibodies. 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 monoclonal antibody 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 monoclonal antibodies 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 monoclonal antibodies in the same patient group, the association was not significant.

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


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