Abstract
The human pre-B acute lymphoblastic leukemia cell line, BLIN-1, has been previously shown to undergo kappa light chain rearrangement in vitro, making it a valuable resource for analyzing pre-B to B cell differentiation. We have examined the recombination potential of BLIN-1 by characterizing several independently derived kappa-expressing subclones for DNA rearrangement and V kappa gene usage. Analysis of five kappa-expressing subclones (all having the same heavy chain rearrangement) demonstrated independent kappa light chain rearrangement events by DNA hybridization analysis. Northern blot analysis using probes recognizing the four different V kappa families revealed that two subclones used the most proximal V kappa (V kappa IV), one subclone used a V kappa I, and one subclone used a V kappa II. By polymerase chain reaction analyses, we detected transcripts from rearranged V-J-C kappa genes as well as transcripts from germline J-C kappa and V kappa in BLIN-1 cells induced to rearrange the kappa locus. kappa germline transcripts were also detected in normal developing B cell populations in fetal liver and bone marrow. Our collective results indicate that: (a) BLIN-1 can be induced to rearrange the kappa locus, and this correlates with the expression of germline kappa locus transcripts that may play a role in activating or targeting gene rearrangement; and (b) active rearrangement and usage of V genes representing different kappa families suggest that, like in the mouse, repertoire diversification in humans occurs in the presence of a fixed heavy chain rearrangement.
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- 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]
- Alwine J. C., Kemp D. J., Parker B. A., Reiser J., Renart J., Stark G. R., Wahl G. M. Detection of specific RNAs or specific fragments of DNA by fractionation in gels and transfer to diazobenzyloxymethyl paper. Methods Enzymol. 1979;68:220–242. doi: 10.1016/0076-6879(79)68017-5. [DOI] [PubMed] [Google Scholar]
- Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Calame K., Eaton S. Transcriptional controlling elements in the immunoglobulin and T cell receptor loci. Adv Immunol. 1988;43:235–275. doi: 10.1016/s0065-2776(08)60367-3. [DOI] [PubMed] [Google Scholar]
- 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]
- Collier D. A., Griffin J. A., Wells R. D. Non-B right-handed DNA conformations of homopurine.homopyrimidine sequences in the murine immunoglobulin C alpha switch region. J Biol Chem. 1988 May 25;263(15):7397–7405. [PubMed] [Google Scholar]
- Cuisinier A. M., Fumoux F., Moinier D., Boubli L., Guigou V., Milili M., Schiff C., Fougereau M., Tonnelle C. Rapid expansion of human immunoglobulin repertoire (VH, V kappa, V lambda) expressed in early fetal bone marrow. New Biol. 1990 Aug;2(8):689–699. [PubMed] [Google Scholar]
- Dosch H. M., Lam P., Hui M. F., Hibi T. Concerted generation of Ig isotype diversity in human fetal bone marrow. J Immunol. 1989 Oct 15;143(8):2464–2469. [PubMed] [Google Scholar]
- Feddersen R. M., Van Ness B. G. Corrective recombination of mouse immunoglobulin kappa alleles in Abelson murine leukemia virus-transformed pre-B cells. Mol Cell Biol. 1990 Feb;10(2):569–576. doi: 10.1128/mcb.10.2.569. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gathings W. E., Lawton A. R., Cooper M. D. Immunofluorescent studies of the development of pre-B cells, B lymphocytes and immunoglobulin isotype diversity in humans. Eur J Immunol. 1977 Nov;7(11):804–810. doi: 10.1002/eji.1830071112. [DOI] [PubMed] [Google Scholar]
- Greenstein J. L., Solomon A., Abraham G. N. Monoclonal antibodies reactive with idiotypic and variable-region specific determinants on human immunoglobulins. Immunology. 1984 Jan;51(1):17–25. [PMC free article] [PubMed] [Google Scholar]
- Hieter P. A., Maizel J. V., Jr, Leder P. Evolution of human immunoglobulin kappa J region genes. J Biol Chem. 1982 Feb 10;257(3):1516–1522. [PubMed] [Google Scholar]
- Jaenichen H. R., Pech M., Lindenmaier W., Wildgruber N., Zachau H. G. Composite human VK genes and a model of their evolution. Nucleic Acids Res. 1984 Jul 11;12(13):5249–5263. doi: 10.1093/nar/12.13.5249. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jeffreys A. J., Flavell R. A. A physical map of the DNA regions flanking the rabbit beta-globin gene. Cell. 1977 Oct;12(2):429–439. doi: 10.1016/0092-8674(77)90119-2. [DOI] [PubMed] [Google Scholar]
- Klobeck H. G., Bornkamm G. W., Combriato G., Mocikat R., Pohlenz H. D., Zachau H. G. Subgroup IV of human immunoglobulin K light chains is encoded by a single germline gene. Nucleic Acids Res. 1985 Sep 25;13(18):6515–6529. doi: 10.1093/nar/13.18.6515. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Klobeck H. G., Meindl A., Combriato G., Solomon A., Zachau H. G. Human immunoglobulin kappa light chain genes of subgroups II and III. Nucleic Acids Res. 1985 Sep 25;13(18):6499–6513. doi: 10.1093/nar/13.18.6499. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Klobeck H. G., Zachau H. G. The human CK gene segment and the kappa deleting element are closely linked. Nucleic Acids Res. 1986 Jun 11;14(11):4591–4603. doi: 10.1093/nar/14.11.4591. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Klobeck H. G., Zimmer F. J., Combriato G., Zachau H. G. Linking of the human immunoglobulin VK and JKCK regions by chromosomal walking. Nucleic Acids Res. 1987 Dec 10;15(23):9655–9665. doi: 10.1093/nar/15.23.9655. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lennon G. G., Perry R. P. The temporal order of appearance of transcripts from unrearranged and rearranged Ig genes in murine fetal liver. J Immunol. 1990 Mar 1;144(5):1983–1987. [PubMed] [Google Scholar]
- Martin D. J., Van Ness B. G. Identification of a germ line transcript from the unrearranged kappa gene in human B cells. Mol Cell Biol. 1989 Oct;9(10):4560–4562. doi: 10.1128/mcb.9.10.4560. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Martin D. J., van Ness B. G. Initiation and processing of two kappa immunoglobulin germ line transcripts in mouse B cells. Mol Cell Biol. 1990 May;10(5):1950–1958. doi: 10.1128/mcb.10.5.1950. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mather E. L., Perry R. P. Transcriptional regulation of immunoglobulin V genes. Nucleic Acids Res. 1981 Dec 21;9(24):6855–6867. doi: 10.1093/nar/9.24.6855. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nelson K. J., Haimovich J., Perry R. P. Characterization of productive and sterile transcripts from the immunoglobulin heavy-chain locus: processing of micron and muS mRNA. Mol Cell Biol. 1983 Jul;3(7):1317–1332. doi: 10.1128/mcb.3.7.1317. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Reth M. G., Alt F. W. Novel immunoglobulin heavy chains are produced from DJH gene segment rearrangements in lymphoid cells. 1984 Nov 29-Dec 5Nature. 312(5993):418–423. doi: 10.1038/312418a0. [DOI] [PubMed] [Google Scholar]
- Reth M. G., Ammirati P., Jackson S., Alt F. W. Regulated progression of a cultured pre-B-cell line to the B-cell stage. 1985 Sep 26-Oct 2Nature. 317(6035):353–355. doi: 10.1038/317353a0. [DOI] [PubMed] [Google Scholar]
- Schatz D. G., Oettinger M. A., Baltimore D. The V(D)J recombination activating gene, RAG-1. Cell. 1989 Dec 22;59(6):1035–1048. doi: 10.1016/0092-8674(89)90760-5. [DOI] [PubMed] [Google Scholar]
- Schlissel M. S., Baltimore D. Activation of immunoglobulin kappa gene rearrangement correlates with induction of germline kappa gene transcription. Cell. 1989 Sep 8;58(5):1001–1007. doi: 10.1016/0092-8674(89)90951-3. [DOI] [PubMed] [Google Scholar]
- Schroeder H. W., Jr, Hillson J. L., Perlmutter R. M. Early restriction of the human antibody repertoire. Science. 1987 Nov 6;238(4828):791–793. doi: 10.1126/science.3118465. [DOI] [PubMed] [Google Scholar]
- Van Ness B. G., Weigert M., Coleclough C., Mather E. L., Kelley D. E., Perry R. P. Transcription of the unrearranged mouse C kappa locus: sequence of the initiation region and comparison of activity with a rearranged V kappa-C kappa gene. Cell. 1981 Dec;27(3 Pt 2):593–602. doi: 10.1016/0092-8674(81)90401-3. [DOI] [PubMed] [Google Scholar]
- Wörmann B., Anderson J. M., Liberty J. A., Gajl-Peczalska K., Brunning R. D., Silberman T. L., Arthur D. C., LeBien T. W. Establishment of a leukemic cell model for studying human pre-B to B cell differentiation. J Immunol. 1989 Jan 1;142(1):110–117. [PubMed] [Google Scholar]
- Yancopoulos G. D., Alt F. W. Developmentally controlled and tissue-specific expression of unrearranged VH gene segments. Cell. 1985 Feb;40(2):271–281. doi: 10.1016/0092-8674(85)90141-2. [DOI] [PubMed] [Google Scholar]
- Yancopoulos G. D., Desiderio S. V., Paskind M., Kearney J. F., Baltimore D., Alt F. W. Preferential utilization of the most JH-proximal VH gene segments in pre-B-cell lines. Nature. 1984 Oct 25;311(5988):727–733. doi: 10.1038/311727a0. [DOI] [PubMed] [Google Scholar]