Genetic immunization: a new monoclonal antibody for the detection of BCL-6 protein in paraffin sections

J Histochem Cytochem. 2006 Jan;54(1):31-8. doi: 10.1369/jhc.5A6646.2005. Epub 2005 Jul 26.

Abstract

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.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal*
  • Cell Line, Tumor
  • DNA-Binding Proteins / immunology
  • DNA-Binding Proteins / metabolism*
  • Diagnosis, Differential
  • Fixatives
  • Formaldehyde
  • Humans
  • Immunohistochemistry
  • Leukemia, Lymphocytic, Chronic, B-Cell / diagnosis
  • Leukemia, Lymphocytic, Chronic, B-Cell / metabolism
  • Lymphoma, Non-Hodgkin / diagnosis
  • Lymphoma, Non-Hodgkin / metabolism
  • Mice
  • Mice, Inbred BALB C
  • Palatine Tonsil / metabolism
  • Paraffin Embedding
  • Proto-Oncogene Proteins c-bcl-6
  • Survival Analysis
  • Tissue Array Analysis

Substances

  • Antibodies, Monoclonal
  • BCL6 protein, human
  • DNA-Binding Proteins
  • Fixatives
  • Proto-Oncogene Proteins c-bcl-6
  • Formaldehyde