Skip to main content
The American Journal of Pathology logoLink to The American Journal of Pathology
. 1995 Aug;147(2):405–411.

A Specific Monoclonal Antibody (PG-B6) Detects Expression of the BCL-6 Protein in Germinal Center B Cells

Leonardo Flenghi, Bihui H Ye, Marco Fizzotti, Barbara Bigerna, Giorgio Cattoretti, Stefania Venturi, Roberta Pacini, Stefano Pileri, Francesco Lo Coco, Edoardo Pescarmona, Pier-Giuseppe Pelicci, Riccardo Dalla-Favera, Brunangelo Falini
PMCID: PMC1869830  PMID: 7639334

Abstract

The BCL-6 gene is frequently involved in translocations occurring at the 3q27 locus and is rear-ranged in approximately 30% of diffuse large cell lymphomas and in a small fraction of follicular lymphomas. The BCL-6 gene encodes for a Kruppel-type zinc-finger protein, the cell/tissue expression and function of which is unknown. In this study, we describe a new monoclonal anti-body (PG-B6) that is specificaly directed against a fixative-sensitive epitope on the amino-terminal region of the BCL-6protein. By immunocytochemical analysis, BCL-6 localizes in the nucleus where PG-B6 staining gives a microgranular/diffuse pattern with exclusion of the nucleoli. The main reactivity of PG-B6in tonsil and spleen is with the nuclei of germinal center B cells, whereas B cells within the mantle and marginal zones do not express BCL-6. No other lymphoid cells in the tonsil express BCL-6 except for a subset of CD3+/CD4+ intrafollicular and interfollicular T cells. A few lymphoid cells of unknown phenotype express BCL-6 in the thymus. Extra-lymphoid BCL-6 expression includes a weak nuclear positivity of epithelia. In non-Hodgkin's lymphomas, BCL-6 expression parallels that observed in normal lymphoid compartments, eg, expression in germinal center-derived tumors (follicular and diffuse large cell lymphomas), but not in mantle cell and marginal zone lymphomas. In most diffuse large cell lymphomas, the BCL-6 protein is expressed at high levels in cases with or without BCL-6 gene rearrangements. These findings indicate that BCL-6 expression is specifically regulated during B lymphocyte development and suggest that BCL-6 may play a role during B cell differentiation in the germinal center.

Full text

PDF
405

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Baron B. W., Nucifora G., McCabe N., Espinosa R., 3rd, Le Beau M. M., McKeithan T. W. Identification of the gene associated with the recurring chromosomal translocations t(3;14)(q27;q32) and t(3;22)(q27;q11) in B-cell lymphomas. Proc Natl Acad Sci U S A. 1993 Jun 1;90(11):5262–5266. doi: 10.1073/pnas.90.11.5262. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bastard C., Deweindt C., Kerckaert J. P., Lenormand B., Rossi A., Pezzella F., Fruchart C., Duval C., Monconduit M., Tilly H. LAZ3 rearrangements in non-Hodgkin's lymphoma: correlation with histology, immunophenotype, karyotype, and clinical outcome in 217 patients. Blood. 1994 May 1;83(9):2423–2427. [PubMed] [Google Scholar]
  3. Cordell J. L., Falini B., Erber W. N., Ghosh A. K., Abdulaziz Z., MacDonald S., Pulford K. A., Stein H., Mason D. Y. Immunoenzymatic labeling of monoclonal antibodies using immune complexes of alkaline phosphatase and monoclonal anti-alkaline phosphatase (APAAP complexes). J Histochem Cytochem. 1984 Feb;32(2):219–229. doi: 10.1177/32.2.6198355. [DOI] [PubMed] [Google Scholar]
  4. Deweindt C., Kerckaert J. P., Tilly H., Quief S., Nguyen V. C., Bastard C. Cloning of a breakpoint cluster region at band 3q27 involved in human non-Hodgkin's lymphoma. Genes Chromosomes Cancer. 1993 Nov;8(3):149–154. doi: 10.1002/gcc.2870080303. [DOI] [PubMed] [Google Scholar]
  5. Falini B., Abdulaziz Z., Gerdes J., Canino S., Ciani C., Cordell J. L., Knight P. M., Stein H., Grignani F., Martelli M. F. Description of a sequential staining procedure for double immunoenzymatic staining of pairs of antigens using monoclonal antibodies. J Immunol Methods. 1986 Nov 6;93(2):265–273. doi: 10.1016/0022-1759(86)90199-7. [DOI] [PubMed] [Google Scholar]
  6. Falini B., Flenghi L., Pileri S., Gambacorta M., Bigerna B., Durkop H., Eitelbach F., Thiele J., Pacini R., Cavaliere A. PG-M1: a new monoclonal antibody directed against a fixative-resistant epitope on the macrophage-restricted form of the CD68 molecule. Am J Pathol. 1993 May;142(5):1359–1372. [PMC free article] [PubMed] [Google Scholar]
  7. Falini B., Pileri S., Martelli M. F. Histological and immunohistological analysis of human lymphomas. Crit Rev Oncol Hematol. 1989;9(4):351–419. doi: 10.1016/s1040-8428(89)80018-6. [DOI] [PubMed] [Google Scholar]
  8. Flenghi L., Fagioli M., Tomassoni L., Pileri S., Gambacorta M., Pacini R., Grignani F., Casini T., Ferrucci P. F., Martelli M. F. Characterization of a new monoclonal antibody (PG-M3) directed against the aminoterminal portion of the PML gene product: immunocytochemical evidence for high expression of PML proteins on activated macrophages, endothelial cells, and epithelia. Blood. 1995 Apr 1;85(7):1871–1880. [PubMed] [Google Scholar]
  9. Harris N. L., Jaffe E. S., Stein H., Banks P. M., Chan J. K., Cleary M. L., Delsol G., De Wolf-Peeters C., Falini B., Gatter K. C. A revised European-American classification of lymphoid neoplasms: a proposal from the International Lymphoma Study Group. Blood. 1994 Sep 1;84(5):1361–1392. [PubMed] [Google Scholar]
  10. Kerckaert J. P., Deweindt C., Tilly H., Quief S., Lecocq G., Bastard C. LAZ3, a novel zinc-finger encoding gene, is disrupted by recurring chromosome 3q27 translocations in human lymphomas. Nat Genet. 1993 Sep;5(1):66–70. doi: 10.1038/ng0993-66. [DOI] [PubMed] [Google Scholar]
  11. Liu Y. J., Johnson G. D., Gordon J., MacLennan I. C. Germinal centres in T-cell-dependent antibody responses. Immunol Today. 1992 Jan;13(1):17–21. doi: 10.1016/0167-5699(92)90199-H. [DOI] [PubMed] [Google Scholar]
  12. Lo Coco F., Ye B. H., Lista F., Corradini P., Offit K., Knowles D. M., Chaganti R. S., Dalla-Favera R. Rearrangements of the BCL6 gene in diffuse large cell non-Hodgkin's lymphoma. Blood. 1994 Apr 1;83(7):1757–1759. [PubMed] [Google Scholar]
  13. Mason D. Y., Cordell J. L., Abdulaziz Z., Naiem M., Bordenave G. Preparation of peroxidase: antiperoxidase (PAP) complexes for immunohistological labeling of monoclonal antibodies. J Histochem Cytochem. 1982 Nov;30(11):1114–1122. doi: 10.1177/30.11.6183312. [DOI] [PubMed] [Google Scholar]
  14. Miki T., Kawamata N., Hirosawa S., Aoki N. Gene involved in the 3q27 translocation associated with B-cell lymphoma, BCL5, encodes a Krüppel-like zinc-finger protein. Blood. 1994 Jan 1;83(1):26–32. [PubMed] [Google Scholar]
  15. Offit K., Jhanwar S., Ebrahim S. A., Filippa D., Clarkson B. D., Chaganti R. S. t(3;22)(q27;q11): a novel translocation associated with diffuse non-Hodgkin's lymphoma. Blood. 1989 Nov 1;74(6):1876–1879. [PubMed] [Google Scholar]
  16. Offit K., Lo Coco F., Louie D. C., Parsa N. Z., Leung D., Portlock C., Ye B. H., Lista F., Filippa D. A., Rosenbaum A. Rearrangement of the bcl-6 gene as a prognostic marker in diffuse large-cell lymphoma. N Engl J Med. 1994 Jul 14;331(2):74–80. doi: 10.1056/NEJM199407143310202. [DOI] [PubMed] [Google Scholar]
  17. Ruppert J. M., Kinzler K. W., Wong A. J., Bigner S. H., Kao F. T., Law M. L., Seuanez H. N., O'Brien S. J., Vogelstein B. The GLI-Kruppel family of human genes. Mol Cell Biol. 1988 Aug;8(8):3104–3113. doi: 10.1128/mcb.8.8.3104. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Thorbecke G. J., Amin A. R., Tsiagbe V. K. Biology of germinal centers in lymphoid tissue. FASEB J. 1994 Aug;8(11):832–840. doi: 10.1096/fasebj.8.11.8070632. [DOI] [PubMed] [Google Scholar]
  19. Weis K., Rambaud S., Lavau C., Jansen J., Carvalho T., Carmo-Fonseca M., Lamond A., Dejean A. Retinoic acid regulates aberrant nuclear localization of PML-RAR alpha in acute promyelocytic leukemia cells. Cell. 1994 Jan 28;76(2):345–356. doi: 10.1016/0092-8674(94)90341-7. [DOI] [PubMed] [Google Scholar]
  20. Ye B. H., Lista F., Lo Coco F., Knowles D. M., Offit K., Chaganti R. S., Dalla-Favera R. Alterations of a zinc finger-encoding gene, BCL-6, in diffuse large-cell lymphoma. Science. 1993 Oct 29;262(5134):747–750. doi: 10.1126/science.8235596. [DOI] [PubMed] [Google Scholar]
  21. Ye B. H., Rao P. H., Chaganti R. S., Dalla-Favera R. Cloning of bcl-6, the locus involved in chromosome translocations affecting band 3q27 in B-cell lymphoma. Cancer Res. 1993 Jun 15;53(12):2732–2735. [PubMed] [Google Scholar]

Articles from The American Journal of Pathology are provided here courtesy of American Society for Investigative Pathology

RESOURCES