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. 1993 Sep 1;178(3):1023–1031. doi: 10.1084/jem.178.3.1023

The bone marrow of multiple myeloma patients contains B cell populations at different stages of differentiation that are clonally related to the malignant plasma cell

PMCID: PMC2191154  PMID: 8350044

Abstract

One of the distinguishing features of multiple myeloma (MM) is the proliferation of a clonal plasma cell population in the bone marrow (BM). It is of particular interest that the tumor plasma cells appear to be restricted to the microenvironment of the BM and are rarely detected in the peripheral system, yet the disease is found widely disseminated throughout the axial skeleton. Furthermore, isolation of MM tumor cell lines has proven to be quite problematic due to their slow growth rate. These observations have instigated the search for earlier cells in the B cell lineage that are clonally related to the plasma cell tumor and that may represent the growth fraction of the tumor. We used allele-specific oligonucleotides (ASO) derived from the third complementarity determining region of the rearranged tumor immunoglobulin heavy chain gene to detect isotypes clonally related to the plasma cell tumor. By reverse transcribing RNA from the BM with a panel of CH primers (mu, delta, alpha, and gamma), followed by ASO- polymerase chain reaction amplification, we demonstrate the existence of preswitch isotype species that are clonally related to the myeloma tumor. Furthermore, we show that separation of the BM cells into CD45+ and CD38+ cell populations results in a lineage-specific expression of the clonally related RNA molecules, with the C mu and C delta in the CD45+, and C gamma in the CD38+ population. Interestingly, clonally related C alpha transcripts are also derived from the CD45+ fraction. These results confirm the presence of B cell populations clonally related to the plasma cell tumor and are consistent with models that propose the existence of myeloma precursors.

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Selected References

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  1. Bast E. J., van Camp B., Reynaert P., Wiringa G., Ballieux R. E. Idiotypic peripheral blood lymphocytes in monoclonal gammopathy. Clin Exp Immunol. 1982 Mar;47(3):677–682. [PMC free article] [PubMed] [Google Scholar]
  2. Berenson J., Wong R., Kim K., Brown N., Lichtenstein A. Evidence for peripheral blood B lymphocyte but not T lymphocyte involvement in multiple myeloma. Blood. 1987 Nov;70(5):1550–1553. [PubMed] [Google Scholar]
  3. Bergui L., Schena M., Gaidano G., Riva M., Caligaris-Cappio F. Interleukin 3 and interleukin 6 synergistically promote the proliferation and differentiation of malignant plasma cell precursors in multiple myeloma. J Exp Med. 1989 Aug 1;170(2):613–618. doi: 10.1084/jem.170.2.613. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bhan A. K., Nadler L. M., Stashenko P., McCluskey R. T., Schlossman S. F. Stages of B cell differentiation in human lymphoid tissue. J Exp Med. 1981 Sep 1;154(3):737–749. doi: 10.1084/jem.154.3.737. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Billadeau D., Blackstadt M., Greipp P., Kyle R. A., Oken M. M., Kay N., Van Ness B. Analysis of B-lymphoid malignancies using allele-specific polymerase chain reaction: a technique for sequential quantitation of residual disease. Blood. 1991 Dec 1;78(11):3021–3029. [PubMed] [Google Scholar]
  6. Billadeau D., Quam L., Thomas W., Kay N., Greipp P., Kyle R., Oken M. M., Van Ness B. Detection and quantitation of malignant cells in the peripheral blood of multiple myeloma patients. Blood. 1992 Oct 1;80(7):1818–1824. [PubMed] [Google Scholar]
  7. Boccadoro M., Van Acker A., Pileri A., Urbain J. Idiotypic lymphocytes in human monoclonal gammopathies. Ann Immunol (Paris) 1981 Jan-Feb;132C(1):9–19. doi: 10.1016/0769-2625(81)90003-9. [DOI] [PubMed] [Google Scholar]
  8. Caligaris-Cappio F., Bergui L., Tesio L., Pizzolo G., Malavasi F., Chilosi M., Campana D., van Camp B., Janossy G. Identification of malignant plasma cell precursors in the bone marrow of multiple myeloma. J Clin Invest. 1985 Sep;76(3):1243–1251. doi: 10.1172/JCI112080. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cassel A., Leibovitz N., Hornstein L., Quitt M., Aghai E. Evidence for the existence of circulating monoclonal B-lymphocytes in multiple myeloma patients. Exp Hematol. 1990 Dec;18(11):1171–1173. [PubMed] [Google Scholar]
  10. Chan C. S., Wormsley S. B., Pierce L. E., Peter J. B., Schechter G. P. B-cell surface phenotypes of proliferating myeloma cells: target antigens for immunotherapy. Am J Hematol. 1990 Feb;33(2):101–109. doi: 10.1002/ajh.2830330206. [DOI] [PubMed] [Google Scholar]
  11. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  12. Clofent G., Klein B., Commes T., Ghanem N., Lefranc M. P., Bataille R. No detectable malignant B cells in the peripheral blood of patients with multiple myeloma. Br J Haematol. 1989 Mar;71(3):357–361. doi: 10.1111/j.1365-2141.1989.tb04292.x. [DOI] [PubMed] [Google Scholar]
  13. Clofent G., Klein B., Commes T., Vincent C., Ghanem N., Lenoir G., Lefranc M. P., Bataille R. Limiting dilution cloning of B cells from patients with multiple myeloma: emergence of non-malignant B-cell lines. Int J Cancer. 1989 Apr 15;43(4):578–586. doi: 10.1002/ijc.2910430408. [DOI] [PubMed] [Google Scholar]
  14. Deane M., McCarthy K. P., Wiedemann L. M., Norton J. D. An improved method for detection of B-lymphoid clonality by polymerase chain reaction. Leukemia. 1991 Aug;5(8):726–730. [PubMed] [Google Scholar]
  15. Deane M., Norton J. D. Immunoglobulin heavy chain variable region family usage is independent of tumor cell phenotype in human B lineage leukemias. Eur J Immunol. 1990 Oct;20(10):2209–2217. doi: 10.1002/eji.1830201009. [DOI] [PubMed] [Google Scholar]
  16. Epstein J., Barlogie B., Katzmann J., Alexanian R. Phenotypic heterogeneity in aneuploid multiple myeloma indicates pre-B cell involvement. Blood. 1988 Apr;71(4):861–865. [PubMed] [Google Scholar]
  17. Epstein J., Xiao H. Q., He X. Y. Markers of multiple hematopoietic-cell lineages in multiple myeloma. N Engl J Med. 1990 Mar 8;322(10):664–668. doi: 10.1056/NEJM199003083221005. [DOI] [PubMed] [Google Scholar]
  18. Grogan T. M., Durie B. G., Lomen C., Spier C., Wirt D. P., Nagle R., Wilson G. S., Richter L., Vela E., Maxey V. Delineation of a novel pre-B cell component in plasma cell myeloma: immunochemical, immunophenotypic, genotypic, cytologic, cell culture, and kinetic features. Blood. 1987 Oct;70(4):932–942. [PubMed] [Google Scholar]
  19. Grogan T. M., Durie B. G., Spier C. M., Richter L., Vela E. Myelomonocytic antigen positive multiple myeloma. Blood. 1989 Feb 15;73(3):763–769. [PubMed] [Google Scholar]
  20. Ichihara Y., Matsuoka H., Kurosawa Y. Organization of human immunoglobulin heavy chain diversity gene loci. EMBO J. 1988 Dec 20;7(13):4141–4150. doi: 10.1002/j.1460-2075.1988.tb03309.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Jackson D. G., Bell J. I. Isolation of a cDNA encoding the human CD38 (T10) molecule, a cell surface glycoprotein with an unusual discontinuous pattern of expression during lymphocyte differentiation. J Immunol. 1990 Apr 1;144(7):2811–2815. [PubMed] [Google Scholar]
  22. Jensen G. S., Mant M. J., Belch A. J., Berenson J. R., Ruether B. A., Pilarski L. M. Selective expression of CD45 isoforms defines CALLA+ monoclonal B-lineage cells in peripheral blood from myeloma patients as late stage B cells. Blood. 1991 Aug 1;78(3):711–719. [PubMed] [Google Scholar]
  23. Jonsson O. G., Kitchens R. L., Scott F. C., Smith R. G. Detection of minimal residual disease in acute lymphoblastic leukemia using immunoglobulin hypervariable region specific oligonucleotide probes. Blood. 1990 Nov 15;76(10):2072–2079. [PubMed] [Google Scholar]
  24. King M. A., Nelson D. S. Tumor cell heterogeneity in multiple myeloma: antigenic, morphologic, and functional studies of cells from blood and bone marrow. Blood. 1989 May 15;73(7):1925–1935. [PubMed] [Google Scholar]
  25. Klein D., Nietupski J., Sirlin S., Stavnezer J. I.29 lymphoma cells express a nonmutated VH gene before and after H chain switch. J Immunol. 1988 Mar 1;140(5):1676–1684. [PubMed] [Google Scholar]
  26. Kubagawa H., Vogler L. B., Capra J. D., Conrad M. E., Lawton A. R., Cooper M. D. Studies on the clonal origin of multiple myeloma. Use of individually specific (idiotype) antibodies to trace the oncogenic event to its earliest point of expression in B-cell differentiation. J Exp Med. 1979 Oct 1;150(4):792–807. doi: 10.1084/jem.150.4.792. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Levy Y., Schmitt C., Tsapis A., Brouet J. C., Fermand J. P. Phenotype and immunoglobulin gene configuration of blood B cells from patients with multiple myeloma. Clin Exp Immunol. 1991 Jun;84(3):435–439. [PMC free article] [PubMed] [Google Scholar]
  28. Perlmutter A. P., Gilbert W. Antibodies of the secondary response can be expressed without switch recombination in normal mouse B cells. Proc Natl Acad Sci U S A. 1984 Nov;81(22):7189–7193. doi: 10.1073/pnas.81.22.7189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Ravetch J. V., Siebenlist U., Korsmeyer S., Waldmann T., Leder P. Structure of the human immunoglobulin mu locus: characterization of embryonic and rearranged J and D genes. Cell. 1981 Dec;27(3 Pt 2):583–591. doi: 10.1016/0092-8674(81)90400-1. [DOI] [PubMed] [Google Scholar]
  30. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  31. Shimizu A., Nussenzweig M. C., Han H., Sanchez M., Honjo T. Trans-splicing as a possible molecular mechanism for the multiple isotype expression of the immunoglobulin gene. J Exp Med. 1991 Jun 1;173(6):1385–1393. doi: 10.1084/jem.173.6.1385. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Shockett P., Stavnezer J. Effect of cytokines on switching to IgA and alpha germline transcripts in the B lymphoma I.29 mu. Transforming growth factor-beta activates transcription of the unrearranged C alpha gene. J Immunol. 1991 Dec 15;147(12):4374–4383. [PubMed] [Google Scholar]
  33. Terstappen L. W., Johnsen S., Segers-Nolten I. M., Loken M. R. Identification and characterization of plasma cells in normal human bone marrow by high-resolution flow cytometry. Blood. 1990 Nov 1;76(9):1739–1747. [PubMed] [Google Scholar]
  34. Van 't Veer M. B., Radl J., Waltman F. L., Moberts M., Bolhuis R. L. Simultaneous expression of cytoplasmic IgM and IgG in plasma cells in a patient with IgG paraproteinaemia. Eur J Haematol. 1989 Aug;43(2):178–181. doi: 10.1111/j.1600-0609.1989.tb00275.x. [DOI] [PubMed] [Google Scholar]
  35. Van Riet I., Heirman C., Lacor P., De Waele M., Thielemans K., Van Camp B. Detection of monoclonal B lymphocytes in bone marrow and peripheral blood of multiple myeloma patients by immunoglobulin gene rearrangement studies. Br J Haematol. 1989 Nov;73(3):289–295. doi: 10.1111/j.1365-2141.1989.tb07742.x. [DOI] [PubMed] [Google Scholar]
  36. 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]
  37. Yamada M., Wasserman R., Lange B., Reichard B. A., Womer R. B., Rovera G. Minimal residual disease in childhood B-lineage lymphoblastic leukemia. Persistence of leukemic cells during the first 18 months of treatment. N Engl J Med. 1990 Aug 16;323(7):448–455. doi: 10.1056/NEJM199008163230705. [DOI] [PubMed] [Google Scholar]

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