Abstract
A method was established to detect clonal lambda light chain gene rearrangements in peripheral blood lymphocytes and frozen or paraffin-embedded tissues. V lambda-gene-family-specific primers were used together with a J lambda primer mix in separate reactions to amplify V lambda gene rearrangements by the polymerase chain reaction. Clonal lambda gene rearrangements were detected in seven of seven lambda-expressing B cell leukemias, in four of five lambda-expressing non-Hodgkin's lymphomas with frozen tissues, and in seven of nine cases of lambda-expressing non-Hodgkin's lymphomas for which formalin-fixed, paraffin-embedded specimens were available. Clonality of amplified polymerase chain reaction products was confirmed by sequence analysis for several cases. The present study shows that it is possible to amplify clonal lambda gene rearrangements in the majority of lambda-expressing B cell leukemias and lymphomas. The method described here, therefore, is a useful supplement to the previously described approach of VH and VK gene amplification to detect clonal B cell populations and allows the study of V lambda gene usage and somatic mutation in lambda-expressing normal and malignant B cells.
Full text
PDF








Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Abe M., Ozaki S., Wolfenbarger D., deBram-Hart M., Weiss D. T., Solomon A. Variable-region subgroup distribution among lambda-type immunoglobulins in normal human serum. J Clin Lab Anal. 1994;8(1):4–9. doi: 10.1002/jcla.1860080103. [DOI] [PubMed] [Google Scholar]
- Alt F. W., Blackwell T. K., DePinho R. A., Reth M. G., Yancopoulos G. D. Regulation of genome rearrangement events during lymphocyte differentiation. Immunol Rev. 1986 Feb;89:5–30. doi: 10.1111/j.1600-065x.1986.tb01470.x. [DOI] [PubMed] [Google Scholar]
- Anderson M. L., Szajnert M. F., Kaplan J. C., McColl L., Young B. D. The isolation of a human Ig V lambda gene from a recombinant library of chromosome 22 and estimation of its copy number. Nucleic Acids Res. 1984 Sep 11;12(17):6647–6661. doi: 10.1093/nar/12.17.6647. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bauer T. R., Jr, Blomberg B. The human lambda L chain Ig locus. Recharacterization of JC lambda 6 and identification of a functional JC lambda 7. J Immunol. 1991 Apr 15;146(8):2813–2820. [PubMed] [Google Scholar]
- Beishuizen A., Verhoeven M. A., Mol E. J., van Dongen J. J. Detection of immunoglobulin kappa light-chain gene rearrangement patterns by Southern blot analysis. Leukemia. 1994 Dec;8(12):2228–2239. [PubMed] [Google Scholar]
- Ch'ang L. Y., Yen C. P., Besl L., Schell M., Solomon A. Identification and characterization of a functional human Ig V lambda VI germline gene. Mol Immunol. 1994 May;31(7):531–536. doi: 10.1016/0161-5890(94)90040-x. [DOI] [PubMed] [Google Scholar]
- Combriato G., Klobeck H. G. V lambda and J lambda-C lambda gene segments of the human immunoglobulin lambda light chain locus are separated by 14 kb and rearrange by a deletion mechanism. Eur J Immunol. 1991 Jun;21(6):1513–1522. doi: 10.1002/eji.1830210627. [DOI] [PubMed] [Google Scholar]
- Daley M. D., Olee T., Peng H. Q., Soto-Gil R. W., Chen P. P., Siminovitch K. A. Molecular characterization of the human immunoglobulin V lambda I germline gene repertoire. Mol Immunol. 1992 Sep;29(9):1031–1042. doi: 10.1016/0161-5890(92)90034-u. [DOI] [PubMed] [Google Scholar]
- Daley M. D., Peng H. Q., Misener V., Liu X. Y., Chen P. P., Siminovitch K. A. Molecular analysis of human immunoglobulin V lambda germline genes: subgroups V lambda III and V lambda IV. Mol Immunol. 1992 Dec;29(12):1515–1518. doi: 10.1016/0161-5890(92)90226-n. [DOI] [PubMed] [Google Scholar]
- Dariavach P., Lefranc G., Lefranc M. P. Human immunoglobulin C lambda 6 gene encodes the Kern+Oz-lambda chain and C lambda 4 and C lambda 5 are pseudogenes. Proc Natl Acad Sci U S A. 1987 Dec;84(24):9074–9078. doi: 10.1073/pnas.84.24.9074. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Deftos M., Soto-Gil R., Quan M., Olee T., Chen P. P. Utilization of a potentially universal downstream primer in the rapid identification and characterization of V lambda genes from two new human V lambda gene families. Scand J Immunol. 1994 Jan;39(1):95–103. doi: 10.1111/j.1365-3083.1994.tb03345.x. [DOI] [PubMed] [Google Scholar]
- Frippiat J. P., Chuchana P., Bernard F., Buluwela L., Lefranc G., Lefranc M. P. First genomic sequence of a human Ig variable lambda gene belonging to subgroup III. Nucleic Acids Res. 1990 Dec 11;18(23):7134–7134. doi: 10.1093/nar/18.23.7134. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Frippiat J. P., Lefranc M. P. Genomic organisation of 34 kb of the human immunoglobulin lambda locus (IGLV): restriction map and sequences of new V lambda III genes. Mol Immunol. 1994 Jun;31(9):657–670. doi: 10.1016/0161-5890(94)90175-9. [DOI] [PubMed] [Google Scholar]
- Graninger W. B., Goldman P. L., Morton C. C., O'Brien S. J., Korsmeyer S. J. The kappa-deleting element. Germline and rearranged, duplicated and dispersed forms. J Exp Med. 1988 Feb 1;167(2):488–501. doi: 10.1084/jem.167.2.488. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hieter P. A., Hollis G. F., Korsmeyer S. J., Waldmann T. A., Leder P. Clustered arrangement of immunoglobulin lambda constant region genes in man. Nature. 1981 Dec 10;294(5841):536–540. doi: 10.1038/294536a0. [DOI] [PubMed] [Google Scholar]
- Hieter P. A., Korsmeyer S. J., Waldmann T. A., Leder P. Human immunoglobulin kappa light-chain genes are deleted or rearranged in lambda-producing B cells. Nature. 1981 Apr 2;290(5805):368–372. doi: 10.1038/290368a0. [DOI] [PubMed] [Google Scholar]
- Huber C., Klobeck H. G., Zachau H. G. Ongoing V kappa-J kappa recombination after formation of a productive V kappa-J kappa coding joint. Eur J Immunol. 1992 Jun;22(6):1561–1565. doi: 10.1002/eji.1830220632. [DOI] [PubMed] [Google Scholar]
- Irigoyen M., Manheimer-Lory A., Gaynor B., Diamond B. Molecular analysis of the human immunoglobulin V lambda II gene family. J Clin Invest. 1994 Aug;94(2):532–538. doi: 10.1172/JCI117366. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kipps T. J., Tomhave E., Chen P. P., Carson D. A. Autoantibody-associated kappa light chain variable region gene expressed in chronic lymphocytic leukemia with little or no somatic mutation. Implications for etiology and immunotherapy. J Exp Med. 1988 Mar 1;167(3):840–852. doi: 10.1084/jem.167.3.840. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Knowles D. M. Immunophenotypic and immunogenotypic approaches useful in distinguishing benign and malignant lymphoid proliferations. Semin Oncol. 1993 Dec;20(6):583–610. [PubMed] [Google Scholar]
- Korsmeyer S. J., Hieter P. A., Ravetch J. V., Poplack D. G., Waldmann T. A., Leder P. Developmental hierarchy of immunoglobulin gene rearrangements in human leukemic pre-B-cells. Proc Natl Acad Sci U S A. 1981 Nov;78(11):7096–7100. doi: 10.1073/pnas.78.11.7096. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kubagawa H., Cooper M. D., Carroll A. J., Burrows P. D. Light-chain gene expression before heavy-chain gene rearrangement in pre-B cells transformed by Epstein-Barr virus. Proc Natl Acad Sci U S A. 1989 Apr;86(7):2356–2360. doi: 10.1073/pnas.86.7.2356. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Küppers R., Fischer U., Rajewsky K., Gause A. Immunoglobulin heavy and light chain gene sequences of a human CD5 positive immunocytoma and sequences of four novel VHIII germline genes. Immunol Lett. 1992 Sep;34(1):57–62. doi: 10.1016/0165-2478(92)90027-l. [DOI] [PubMed] [Google Scholar]
- Küppers R., Gause A., Rajewsky K. B cells of chronic lymphatic leukemia express V genes in unmutated form. Leuk Res. 1991;15(6):487–496. doi: 10.1016/0145-2126(91)90060-7. [DOI] [PubMed] [Google Scholar]
- Küppers R., Zhao M., Rajewsky K., Hansmann M. L. Detection of clonal B cell populations in paraffin-embedded tissues by polymerase chain reaction. Am J Pathol. 1993 Jul;143(1):230–239. [PMC free article] [PubMed] [Google Scholar]
- Lai E., Wilson R. K., Hood L. E. Physical maps of the mouse and human immunoglobulin-like loci. Adv Immunol. 1989;46:1–59. doi: 10.1016/s0065-2776(08)60650-1. [DOI] [PubMed] [Google Scholar]
- Lenormand B., Ghanem N., Tilly H., Boussemart C., Monconduit M., Piguet H., Lefranc G., Lefranc M. P. Rearrangements of immunoglobulin light and heavy chain genes and correlation with phenotypic markers in B-cell chronic lymphocytic leukemia. Leukemia. 1991 Nov;5(11):928–936. [PubMed] [Google Scholar]
- McCarthy K. P., Sloane J. P., Wiedemann L. M. Rapid method for distinguishing clonal from polyclonal B cell populations in surgical biopsy specimens. J Clin Pathol. 1990 May;43(5):429–432. doi: 10.1136/jcp.43.5.429. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ozaki S., Abe M., Wolfenbarger D., Weiss D. T., Solomon A. Preferential expression of human lambda-light-chain variable-region subgroups in multiple myeloma, AL amyloidosis, and Waldenström's macroglobulinemia. Clin Immunol Immunopathol. 1994 May;71(2):183–189. doi: 10.1006/clin.1994.1070. [DOI] [PubMed] [Google Scholar]
- Potter M. N., Steward C. G., Maitland N. J., Oakhill A. Detection of clonality in childhood B-lineage acute lymphoblastic leukaemia by the polymerase chain reaction. Leukemia. 1992 Apr;6(4):289–294. [PubMed] [Google Scholar]
- Rassenti L. Z., Kipps T. J. Lack of extensive mutations in the VH5 genes used in common B cell chronic lymphocytic leukemia. J Exp Med. 1993 Apr 1;177(4):1039–1046. doi: 10.1084/jem.177.4.1039. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Reed T. J., Reid A., Wallberg K., O'Leary T. J., Frizzera G. Determination of B-cell clonality in paraffin-embedded lymph nodes using the polymerase chain reaction. Diagn Mol Pathol. 1993 Mar;2(1):42–49. [PubMed] [Google Scholar]
- Siminovitch K. A., Bakhshi A., Goldman P., Korsmeyer S. J. A uniform deleting element mediates the loss of kappa genes in human B cells. Nature. 1985 Jul 18;316(6025):260–262. doi: 10.1038/316260a0. [DOI] [PubMed] [Google Scholar]
- Solomon A., Weiss D. T., Murphy C., Fu S. M., Robbins D. L. Chemical and serologic characterization of human lambda VIII light chains. J Immunol. 1994 Aug 15;153(4):1658–1664. [PubMed] [Google Scholar]
- Stiernholm N. B., Kuzniar B., Berinstein N. L. Identification of a new human V lambda gene family--V lambda X. J Immunol. 1994 May 15;152(10):4969–4975. [PubMed] [Google Scholar]
- 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]
- Tsunetsugu-Yokota Y., Minekawa T., Shigemoto K., Shirasawa T., Takemori T. Characterization of a new subgroup of human Ig V lambda cDNA clone and its expression. Mol Immunol. 1992 Jun;29(6):723–728. doi: 10.1016/0161-5890(92)90182-w. [DOI] [PubMed] [Google Scholar]
- Udey J. A., Blomberg B. Human lambda light chain locus: organization and DNA sequences of three genomic J regions. Immunogenetics. 1987;25(1):63–70. doi: 10.1007/BF00768834. [DOI] [PubMed] [Google Scholar]
- Vasicek T. J., Leder P. Structure and expression of the human immunoglobulin lambda genes. J Exp Med. 1990 Aug 1;172(2):609–620. doi: 10.1084/jem.172.2.609. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Williams S. C., Winter G. Cloning and sequencing of human immunoglobulin V lambda gene segments. Eur J Immunol. 1993 Jul;23(7):1456–1461. doi: 10.1002/eji.1830230709. [DOI] [PubMed] [Google Scholar]
- Winkler T. H., Fehr H., Kalden J. R. Analysis of immunoglobulin variable region genes from human IgG anti-DNA hybridomas. Eur J Immunol. 1992 Jul;22(7):1719–1728. doi: 10.1002/eji.1830220709. [DOI] [PubMed] [Google Scholar]
- Zou Y. R., Takeda S., Rajewsky K. Gene targeting in the Ig kappa locus: efficient generation of lambda chain-expressing B cells, independent of gene rearrangements in Ig kappa. EMBO J. 1993 Mar;12(3):811–820. doi: 10.1002/j.1460-2075.1993.tb05721.x. [DOI] [PMC free article] [PubMed] [Google Scholar]