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. 2000 Nov;53(11):835–840. doi: 10.1136/jcp.53.11.835

Evaluation of B cell lymphoid infiltrates in bone marrow biopsies by morphology, immunohistochemistry, and molecular analysis

B Maes 1, R Achten 1, A Demunter 1, B Peeters 1, G Verhoef 1, C De Wolf-Peeters 1
PMCID: PMC1731107  PMID: 11127265

Abstract

Aims—Morphological criteria to distinguish between reactive and neoplastic B cell lymphocytoid infiltrates in trephines have been defined but are not always reliable. Polymerase chain reaction (PCR) analysis of the CDR3 region of the immunoglobulin heavy chain (IgH) gene which, by demonstrating monoclonality, can provide additional arguments in favour of lymphoid malignancy is now frequently used for the detection and follow up of B cell lymphoma (NHL). The aim of this study was to investigate the usefulness of morphological findings in bone marrow biopsies in comparison with data obtained by PCR analysis.

Methods—Eighty nine bone marrow biopsies displaying lymphoid infiltrates were evaluated by morphology and immunohistochemistry as well as by CDR3-PCR using consensus framework 3 (FRW3) and JH primers.

Results—The presence of a clonal B cell proliferation was demonstrated by PCR analysis in 45 biopsies, including 21 samples considered to be positive, 17 to be suspicious, and seven to be negative by morphology. In the remaining 44 trephines we found no evidence of clonality, although 12 of these trephines were thought to be positive by morphology.

Conclusions—These results, revealing an incomplete correlation between CDR3-PCR data and immunomorphological findings, indicate that molecular analysis may be more sensitive and specific in general. However, false negative PCR results do occur, which emphasises the necessity to combine both diagnostic tools in the evaluation of lymphoid infiltrates.

Key Words: bone marrow biopsy • lymphoid infiltrate • polymerase chain reaction

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Selected References

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  1. Achille A., Scarpa A., Montresor M., Scardoni M., Zamboni G., Chilosi M., Capelli P., Franzin G., Menestrina F. Routine application of polymerase chain reaction in the diagnosis of monoclonality of B-cell lymphoid proliferations. Diagn Mol Pathol. 1995 Mar;4(1):14–24. doi: 10.1097/00019606-199503000-00005. [DOI] [PubMed] [Google Scholar]
  2. Bartl R., Frisch B., Burkhardt R., Jäger K., Pappenberger R., Hoffmann-Fezer G. Lymphoproliferations in the bone marrow: identification and evolution, classification and staging. J Clin Pathol. 1984 Mar;37(3):233–254. doi: 10.1136/jcp.37.3.233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bartl R., Frisch B., Burkhardt R., Kettner G., Mahl G., Fateh-Moghadam A., Sund M. Assessment of bone marrow histology in the malignant lymphomas (non-Hodgkin's): correlation with clinical factors for diagnosis, prognosis, classification and staging. Br J Haematol. 1982 Aug;51(4):511–530. doi: 10.1111/j.1365-2141.1982.tb02815.x. [DOI] [PubMed] [Google Scholar]
  4. Bluth R. F., Casey T. T., McCurley T. L. Differentiation of reactive from neoplastic small-cell lymphoid aggregates in paraffin-embedded marrow particle preparations using L-26 (CD20) and UCHL-1 (CD45RO) monoclonal antibodies. Am J Clin Pathol. 1993 Feb;99(2):150–156. doi: 10.1093/ajcp/99.2.150. [DOI] [PubMed] [Google Scholar]
  5. Bomberger C., Singh-Jairam M., Rodey G., Guerriero A., Yeager A. M., Fleming W. H., Holland H. K., Waller E. K. Lymphoid reconstitution after autologous PBSC transplantation with FACS-sorted CD34+ hematopoietic progenitors. Blood. 1998 Apr 1;91(7):2588–2600. [PubMed] [Google Scholar]
  6. Coad J. E., Olson D. J., Christensen D. R., Lander T. A., Chibbar R., McGlennen R. C., Brunning R. D. Correlation of PCR-detected clonal gene rearrangements with bone marrow morphology in patients with B-lineage lymphomas. Am J Surg Pathol. 1997 Sep;21(9):1047–1056. doi: 10.1097/00000478-199709000-00009. [DOI] [PubMed] [Google Scholar]
  7. Coad JE, Olson DJ, Lander TA, McGLENNEN RC. Molecular Assessment of Clonality in Lymphoproliferative Disorders: I. Immunoglobulin Gene Rearrangements. Mol Diagn. 1996 Dec;1(4):335–355. doi: 10.1054/MODI00100335. [DOI] [PubMed] [Google Scholar]
  8. Conlan M. G., Bast M., Armitage J. O., Weisenburger D. D. Bone marrow involvement by non-Hodgkin's lymphoma: the clinical significance of morphologic discordance between the lymph node and bone marrow. Nebraska Lymphoma Study Group. J Clin Oncol. 1990 Jul;8(7):1163–1172. doi: 10.1200/JCO.1990.8.7.1163. [DOI] [PubMed] [Google Scholar]
  9. Crisan D., Mattson J. C. Discordant morphologic features in bone marrow involvement by malignant lymphomas: use of gene rearrangement patterns for diagnosis. Am J Hematol. 1995 Aug;49(4):299–309. doi: 10.1002/ajh.2830490408. [DOI] [PubMed] [Google Scholar]
  10. Fisher D. E., Jacobson J. O., Ault K. A., Harris N. L. Diffuse large cell lymphoma with discordant bone marrow histology. Clinical features and biological implications. Cancer. 1989 Nov 1;64(9):1879–1887. doi: 10.1002/1097-0142(19891101)64:9<1879::aid-cncr2820640921>3.0.co;2-d. [DOI] [PubMed] [Google Scholar]
  11. Freedman A. S., Boyd A. W., Bieber F. R., Daley J., Rosen K., Horowitz J. C., Levy D. N., Nadler L. M. Normal cellular counterparts of B cell chronic lymphocytic leukemia. Blood. 1987 Aug;70(2):418–427. [PubMed] [Google Scholar]
  12. Gong J. Z., Zheng S., Chiarle R., De Wolf-Peeters C., Palestro G., Frizzera G., Inghirami G. Detection of immunoglobulin kappa light chain rearrangements by polymerase chain reaction. An improved method for detecting clonal B-cell lymphoproliferative disorders. Am J Pathol. 1999 Aug;155(2):355–363. doi: 10.1016/s0002-9440(10)65132-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Hodges G. F., Lenhardt T. M., Cotelingam J. D. Bone marrow involvement in large-cell lymphoma. Prognostic implications of discordant disease. Am J Clin Pathol. 1994 Mar;101(3):305–311. doi: 10.1093/ajcp/101.3.305. [DOI] [PubMed] [Google Scholar]
  15. Horny H. P., Engst U., Walz R. S., Kaiserling E. In situ immunophenotyping of lymphocytes in human bone marrow: an immunohistochemical study. Br J Haematol. 1989 Mar;71(3):313–321. doi: 10.1111/j.1365-2141.1989.tb04286.x. [DOI] [PubMed] [Google Scholar]
  16. Inghirami G., Szabolcs M. J., Yee H. T., Corradini P., Cesarman E., Knowles D. M. Detection of immunoglobulin gene rearrangement of B cell non-Hodgkin's lymphomas and leukemias in fresh, unfixed and formalin-fixed, paraffin-embedded tissue by polymerase chain reaction. Lab Invest. 1993 Jun;68(6):746–757. [PubMed] [Google Scholar]
  17. Kluin P. M., van Krieken J. H., Kleiverda K., Kluin-Nelemans H. C. Discordant morphologic characteristics of B-cell lymphomas in bone marrow and lymph node biopsies. Am J Clin Pathol. 1990 Jul;94(1):59–66. doi: 10.1093/ajcp/94.1.59. [DOI] [PubMed] [Google Scholar]
  18. Kubic V. L., Brunning R. D. Immunohistochemical evaluation of neoplasms in bone marrow biopsies using monoclonal antibodies reactive in paraffin-embedded tissue. Mod Pathol. 1989 Nov;2(6):618–629. [PubMed] [Google Scholar]
  19. Linder J., Ye Y. L., Harrington D. S., Armitage J. O., Weisenburger D. D. Monoclonal antibodies marking T lymphocytes in paraffin-embedded tissue. Am J Pathol. 1987 Apr;127(1):1–8. [PMC free article] [PubMed] [Google Scholar]
  20. Norton A. J., Isaacson P. G. Monoclonal antibody L26: an antibody that is reactive with normal and neoplastic B lymphocytes in routinely fixed and paraffin wax embedded tissues. J Clin Pathol. 1987 Dec;40(12):1405–1412. doi: 10.1136/jcp.40.12.1405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Pileri S., Falini B., Sabattini E., Bigerna B., Gherlinzoni F., Tazzari P. L. Immunohistochemistry of malignant lymphomas. Advantages and limitations of the new monoclonal antibodies working in paraffin sections. Haematologica. 1991 May-Jun;76(3):226–234. [PubMed] [Google Scholar]
  22. Pittaluga S., Tierens A., Dodoo Y. L., Delabie J., De Wolf-Peeters C. How reliable is histologic examination of bone marrow trephine biopsy specimens for the staging of non-Hodgkin lymphoma? A study of hairy cell leukemia and mantle cell lymphoma involvement of the bone marrow trephine specimen by histologic, immunohistochemical, and polymerase chain reaction techniques. Am J Clin Pathol. 1999 Feb;111(2):179–184. doi: 10.1093/ajcp/111.2.179. [DOI] [PubMed] [Google Scholar]
  23. Trainor K. J., Brisco M. J., Story C. J., Morley A. A. Monoclonality in B-lymphoproliferative disorders detected at the DNA level. Blood. 1990 Jun 1;75(11):2220–2222. [PubMed] [Google Scholar]
  24. Waldmann T. A. The arrangement of immunoglobulin and T cell receptor genes in human lymphoproliferative disorders. Adv Immunol. 1987;40:247–321. doi: 10.1016/s0065-2776(08)60241-2. [DOI] [PubMed] [Google Scholar]
  25. Wan J. H., Trainor K. J., Brisco M. J., Morley A. A. Monoclonality in B cell lymphoma detected in paraffin wax embedded sections using the polymerase chain reaction. J Clin Pathol. 1990 Nov;43(11):888–890. doi: 10.1136/jcp.43.11.888. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Yamada M., Hudson S., Tournay O., Bittenbender S., Shane S. S., Lange B., Tsujimoto Y., Caton A. J., Rovera G. Detection of minimal disease in hematopoietic malignancies of the B-cell lineage by using third-complementarity-determining region (CDR-III)-specific probes. Proc Natl Acad Sci U S A. 1989 Jul;86(13):5123–5127. doi: 10.1073/pnas.86.13.5123. [DOI] [PMC free article] [PubMed] [Google Scholar]

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