Abstract
We have attempted to determine whether T cells and B cells that have the same antigenic specificity and whose receptors share idiotypic determinants in fact express similar VH gene segments. To do this, we have obtained and characterized a cDNA clone containing the entire coding sequence for the VH gene from a glutamic acid60/alanine30/tyrosine10 (GAT)-binding immunoglobulin that carries the CGAT idiotype. The GAT-VH clone was hybridized to Northern blots of GAT-specific T cell RNAs; there was no evidence of a T cell transcript that hybridized to the GAT-VH probe. The T cells analyzed included: (a) 10 GAT-binding suppressor T cell hybridomas, 6 of which secreted factors with CGAT idiotypic determinants, (b) one GAT-specific helper T cell hybridoma, and (c) two GAT-specific helper T cell lines grown in the absence of feeder cells. The detection limit of the Northern blot analysis was 1-2 copies of a particular mRNA species per cell for the hybridomas and 5-10 copies per cell for the T cell lines. Therefore, we conclude that T and B lymphocytes responding to GAT do not utilize similar VH gene segments. Furthermore, the presence of idiotypic determinants on T lymphocytes does not necessarily imply close structural similarity between T and B cell antigen receptors.
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- Apte R. N., Eshhar Z., Löwy I., Zinger H., Mozes E. Characteristics of a poly(LTyr,LGlu)-poly(DLAla)--poly(LLys)-specific helper factor derived from a T cell hybridoma. Eur J Immunol. 1981 Nov;11(11):931–936. doi: 10.1002/eji.1830111115. [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]
- Bach B. A., Greene M. I., Benacerraf B., Nisonoff A. Mechanisms of regulation of cell-mediated immunity. IV. Azobenzenearsonate-specific suppressor factor(s) bear cross-reactive idiotypic determinants the expression of which is linked to the heavy-chain allotype linkage group of genes. J Exp Med. 1979 May 1;149(5):1084–1098. doi: 10.1084/jem.149.5.1084. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bernard O., Hozumi N., Tonegawa S. Sequences of mouse immunoglobulin light chain genes before and after somatic changes. Cell. 1978 Dec;15(4):1133–1144. doi: 10.1016/0092-8674(78)90041-7. [DOI] [PubMed] [Google Scholar]
- Binz H., Wigzell H., Bazin H. T-cell idiotypes are linked to immunoglobulin heavy chain genes. Nature. 1976 Dec 16;264(5587):639–642. doi: 10.1038/264639a0. [DOI] [PubMed] [Google Scholar]
- Binz H., Wigzell H. Shared idiotypic determinants on B and T lymphocytes reactive against the same antigenic determinants. V. Biochemical and serological characteristics of naturally occurring, soluble antigen-binding T-lymphocyte-derived molecules. Scand J Immunol. 1976;5(5):559–571. doi: 10.1111/j.1365-3083.1976.tb00311.x. [DOI] [PubMed] [Google Scholar]
- Bishop J. O., Morton J. G., Rosbash M., Richardson M. Three abundance classes in HeLa cell messenger RNA. Nature. 1974 Jul 19;250(463):199–204. doi: 10.1038/250199a0. [DOI] [PubMed] [Google Scholar]
- Buell G. N., Wickens M. P., Payvar F., Schimke R. T. Synthesis of full length cDNAs from four partially purified oviduct mRNAs. J Biol Chem. 1978 Apr 10;253(7):2471–2482. [PubMed] [Google Scholar]
- Calame K., Rogers J., Early P., Davis M., Livant D., Wall R., Hood L. Mouse Cmu heavy chain immunoglobulin gene segment contains three intervening sequences separating domains. Nature. 1980 Apr 3;284(5755):452–455. doi: 10.1038/284452a0. [DOI] [PubMed] [Google Scholar]
- Casadaban M. J., Cohen S. N. Analysis of gene control signals by DNA fusion and cloning in Escherichia coli. J Mol Biol. 1980 Apr;138(2):179–207. doi: 10.1016/0022-2836(80)90283-1. [DOI] [PubMed] [Google Scholar]
- Cerny J., Heusser C., Wallich R., Hammerling G. J., Eardley D. D. Immunoglobulin idiotopes expressed by T cells. I. Expression of distinct idiotopes detected by monoclonal antibodies on antigen-specific suppressor T cells. J Exp Med. 1982 Sep 1;156(3):719–730. doi: 10.1084/jem.156.3.719. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
- Cosman D., Kress M., Khoury G., Jay G. Tissue-specific expression of an unusual H-2 (class I)-related gene. Proc Natl Acad Sci U S A. 1982 Aug;79(16):4947–4951. doi: 10.1073/pnas.79.16.4947. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Crews S., Griffin J., Huang H., Calame K., Hood L. A single VH gene segment encodes the immune response to phosphorylcholine: somatic mutation is correlated with the class of the antibody. Cell. 1981 Jul;25(1):59–66. doi: 10.1016/0092-8674(81)90231-2. [DOI] [PubMed] [Google Scholar]
- Denhardt D. T. A membrane-filter technique for the detection of complementary DNA. Biochem Biophys Res Commun. 1966 Jun 13;23(5):641–646. doi: 10.1016/0006-291x(66)90447-5. [DOI] [PubMed] [Google Scholar]
- Edelman G. M. Summary: understanding selective molecular recognition. Cold Spring Harb Symp Quant Biol. 1977;41(Pt 2):891–902. doi: 10.1101/sqb.1977.041.01.098. [DOI] [PubMed] [Google Scholar]
- Efstratiadis A., Kafatos F. C., Maxam A. M., Maniatis T. Enzymatic in vitro synthesis of globin genes. Cell. 1976 Feb;7(2):279–288. doi: 10.1016/0092-8674(76)90027-1. [DOI] [PubMed] [Google Scholar]
- Eichmann K. Expression and function of idiotypes of lymphocytes. Adv Immunol. 1978;26:195–254. doi: 10.1016/s0065-2776(08)60231-x. [DOI] [PubMed] [Google Scholar]
- Eichmann K. Idiotype suppression. I. Influence of the dose and of the effector functions of anti-idiotypic antibody on the production of an idiotype. Eur J Immunol. 1974 Apr;4(4):296–302. doi: 10.1002/eji.1830040413. [DOI] [PubMed] [Google Scholar]
- Germain R. N., Ju S. T., Kipps T. J., Benacerraf B., Dorf M. E. Shared idiotypic determinants on antibodies and T-cell-derived suppressor factor specific for the random terpolymer L-glutamic acid60-L-alanine30-L-tyrosine10. J Exp Med. 1979 Mar 1;149(3):613–622. doi: 10.1084/jem.149.3.613. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Glimcher L. H., Hamano T., Asofsky R., Sachs D. H., Pierres M., Samelson L. E., Sharrow S. O., Paul W. E. IA mutant functional antigen-presenting cell lines. J Immunol. 1983 May;130(5):2287–2294. [PubMed] [Google Scholar]
- Gougeon M. L., Leclercq L., Löwy I., Bismuth G., Somme G., Theze J. In vitro inhibition of the helper activity of GAT-specific T-cell lines by a syngeneic anti-idiotypic serum: preferential effect on the IgG response. Cell Immunol. 1982 Aug;71(2):254–269. doi: 10.1016/0008-8749(82)90260-x. [DOI] [PubMed] [Google Scholar]
- Harvey M. A., Adorini L., Miller A., Sercarz E. E. Lysozyme-induced T-suppressor cells and antibodies have a predominant idiotype. Nature. 1979 Oct 18;281(5732):594–596. doi: 10.1038/281594a0. [DOI] [PubMed] [Google Scholar]
- Hedrick S. M., Matis L. A., Hecht T. T., Samelson L. E., Longo D. L., Heber-Katz E., Schwartz R. H. The fine specificity of antigen and Ia determinant recognition by T cell hybridoma clones specific for pigeon cytochrome c. Cell. 1982 Aug;30(1):141–152. doi: 10.1016/0092-8674(82)90020-4. [DOI] [PubMed] [Google Scholar]
- Houghton M., Stewart A. G., Doel S. M., Emtage J. S., Eaton M. A., Smith J. C., Patel T. P., Lewis H. M., Porter A. G., Birch J. R. The amino-terminal sequence of human fibroblast interferon as deduced from reverse transcripts obtained using synthetic oligonucleotide primers. Nucleic Acids Res. 1980 May 10;8(9):1913–1931. doi: 10.1093/nar/8.9.1913. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jerne N. K. Towards a network theory of the immune system. Ann Immunol (Paris) 1974 Jan;125C(1-2):373–389. [PubMed] [Google Scholar]
- Ju S. T., Benacerraf B., Dorf M. E. Idiotypic analysis of antibodies to poly(Glu60Ala30Tyr10): interstrain and interspecies idiotypic crossreactions. Proc Natl Acad Sci U S A. 1978 Dec;75(12):6192–6196. doi: 10.1073/pnas.75.12.6192. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kapp J. A., Araneo B. A. Antigen-specific suppressor T cell interactions. I. Induction of an MHC-restricted suppressor factor specific for L-glutamic acid50-L-tyrosine50. J Immunol. 1982 Jun;128(6):2447–2452. [PubMed] [Google Scholar]
- Kapp J. A., Araneo B. A., Clevinger B. L. Suppression of antibody and T cell proliferative responses to L-glutamic acid60-L-alanine30-L-tyrosine10 by a specific monoclonal T cell factor. J Exp Med. 1980 Jul 1;152(1):235–240. doi: 10.1084/jem.152.1.235. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kimoto M., Fathman C. G. Antigen-reactive T cell clones. I. Transcomplementing hybrid I-A-region gene products function effectively in antigen presentation. J Exp Med. 1980 Oct 1;152(4):759–770. doi: 10.1084/jem.152.4.759. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Krawinkel U., Cramer M., Imanishi-Kari T., Jack R. S., Rajewsky K., Mäkelä O. Isolated hapten-binding receptors of sensitized lymphocytes. I. Receptors from nylon wool-enriched mouse T lymphocytes lack serological markers of immunoglobulin constant domains but express heavy chain variable portions. Eur J Immunol. 1977 Aug;7(8):566–573. doi: 10.1002/eji.1830070814. [DOI] [PubMed] [Google Scholar]
- Kronenberg M., Davis M. M., Early P. W., Hood L. E., Watson J. D. Helper and killer T cells do not express B cell immunoglobulin joining and constant region gene segments. J Exp Med. 1980 Dec 1;152(6):1745–1761. doi: 10.1084/jem.152.6.1745. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kronenberg M., Kraig E., Siu G., Kapp J. A., Kappler J., Marrack P., Pierce C. W., Hood L. Three T cell hybridomas do not contain detectable heavy chain variable gene transcripts. J Exp Med. 1983 Jul 1;158(1):210–227. doi: 10.1084/jem.158.1.210. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kurosawa Y., Tonegawa S. Organization, structure, and assembly of immunoglobulin heavy chain diversity DNA segments. J Exp Med. 1982 Jan 1;155(1):201–218. doi: 10.1084/jem.155.1.201. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lane D., Koprowski H. Molecular recognition and the future of monoclonal antibodies. Nature. 1982 Mar 18;296(5854):200–202. doi: 10.1038/296200a0. [DOI] [PubMed] [Google Scholar]
- Lehrach H., Diamond D., Wozney J. M., Boedtker H. RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination. Biochemistry. 1977 Oct 18;16(21):4743–4751. doi: 10.1021/bi00640a033. [DOI] [PubMed] [Google Scholar]
- Lifshitz R., Schwartz M., Mozes E. Linkage of murine (T,G)-A--L-specific idiotypic determinants to the heavy chain constant region allotypic markers. Immunogenetics. 1980;11(2):191–198. doi: 10.1007/BF01567784. [DOI] [PubMed] [Google Scholar]
- Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
- Messing J., Crea R., Seeburg P. H. A system for shotgun DNA sequencing. Nucleic Acids Res. 1981 Jan 24;9(2):309–321. doi: 10.1093/nar/9.2.309. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nakanishi K., Sugimura K., Yaoita Y., Maeda K., Kashiwamura S., Honjo T., Kishimoto T. A T15-idiotype-positive T suppressor hybridoma does not use the T15 VH gene segment. Proc Natl Acad Sci U S A. 1982 Nov;79(22):6984–6988. doi: 10.1073/pnas.79.22.6984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pawlak L. L., Hart D. A., Nisonoff A. Requirements for prolonged suppression of an idiotypic specificity in adult mice. J Exp Med. 1973 Jun 1;137(6):1442–1458. doi: 10.1084/jem.137.6.1442. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Perry R. P., Kelley D. E. Immunoglobulin messenger RNAs in murine cell lines that have characteristics of immature B lymphocytes. Cell. 1979 Dec;18(4):1333–1339. doi: 10.1016/0092-8674(79)90243-5. [DOI] [PubMed] [Google Scholar]
- Petit C., Joskowicz M., Stanislawski M., Thèze J. Genetic control of the immune response to the terpolymer L-glutamic acid 60-L-alanine 30-L-tyrosine10 (GAT). III. Restricted heterogeneity of the anti-GAT response from BALB/c responder mice. Eur J Immunol. 1979 Dec;9(12):922–928. doi: 10.1002/eji.1830091203. [DOI] [PubMed] [Google Scholar]
- Pierres M., Ju S. T., Waltenbaugh C., Dorf M. E., Benacerraf B., Germain R. N. Fine specificity of antibodies to poly(Glu60Ala30Tyr10) produced by hybrid cell lines. Proc Natl Acad Sci U S A. 1979 May;76(5):2425–2429. doi: 10.1073/pnas.76.5.2425. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pillemer E., Weissman I. L. A monoclonal antibody that detects a V kappa-TEPC15 idiotypic determinant cross-reactive with a Thy-1 determinant. J Exp Med. 1981 May 1;153(5):1068–1079. doi: 10.1084/jem.153.5.1068. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
- Roychoudhury R., Wu R. Terminal transferase-catalyzed addition of nucleotides to the 3' termini of DNA. Methods Enzymol. 1980;65(1):43–62. doi: 10.1016/s0076-6879(80)65009-5. [DOI] [PubMed] [Google Scholar]
- Sakano H., Maki R., Kurosawa Y., Roeder W., Tonegawa S. Two types of somatic recombination are necessary for the generation of complete immunoglobulin heavy-chain genes. Nature. 1980 Aug 14;286(5774):676–683. doi: 10.1038/286676a0. [DOI] [PubMed] [Google Scholar]
- Schreiber A. B., Couraud P. O., Andre C., Vray B., Strosberg A. D. Anti-alprenolol anti-idiotypic antibodies bind to beta-adrenergic receptors and modulate catecholamine-sensitive adenylate cyclase. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7385–7389. doi: 10.1073/pnas.77.12.7385. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sege K., Peterson P. A. Use of anti-idiotypic antibodies as cell-surface receptor probes. Proc Natl Acad Sci U S A. 1978 May;75(5):2443–2447. doi: 10.1073/pnas.75.5.2443. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Smith H. O. Recovery of DNA from gels. Methods Enzymol. 1980;65(1):371–380. doi: 10.1016/s0076-6879(80)65048-4. [DOI] [PubMed] [Google Scholar]
- Sorensen C. M., Pierce C. W. Antigen-specific suppression in genetic responder mice to L-glutamic acid60-L-alanine30-L-tyrosine10 (GAT). Characterization of conventional and hybridoma-derived factors produced by suppressor T cells from mice injected as neonates with syngeneic GAT macrophages. J Exp Med. 1982 Dec 1;156(6):1691–1710. doi: 10.1084/jem.156.6.1691. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Steinmetz M., Frelinger J. G., Fisher D., Hunkapiller T., Pereira D., Weissman S. M., Uehara H., Nathenson S., Hood L. Three cDNA clones encoding mouse transplantation antigens: homology to immunoglobulin genes. Cell. 1981 Apr;24(1):125–134. doi: 10.1016/0092-8674(81)90508-0. [DOI] [PubMed] [Google Scholar]
- Steinmetz M., Moore K. W., Frelinger J. G., Sher B. T., Shen F. W., Boyse E. A., Hood L. A pseudogene homologous to mouse transplantation antigens: transplantation antigens are encoded by eight exons that correlate with protein domains. Cell. 1981 Sep;25(3):683–692. doi: 10.1016/0092-8674(81)90175-6. [DOI] [PubMed] [Google Scholar]
- Taylor J. M., Illmensee R., Summers J. Efficeint transcription of RNA into DNA by avian sarcoma virus polymerase. Biochim Biophys Acta. 1976 Sep 6;442(3):324–330. doi: 10.1016/0005-2787(76)90307-5. [DOI] [PubMed] [Google Scholar]
- Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tonnelle C., Pierres M., Ju S. T., Moinier D., Fougereau M. NH2-terminal amino-acid sequences of poly (Glu60, Ala30, Tyr10) (GAT) and poly (Glu60, Ala40) (GA) monoclonal antibody heavy chains. Mol Immunol. 1981 Nov;18(11):979–984. doi: 10.1016/0161-5890(81)90115-2. [DOI] [PubMed] [Google Scholar]
- Villa-Komaroff L., Efstratiadis A., Broome S., Lomedico P., Tizard R., Naber S. P., Chick W. L., Gilbert W. A bacterial clone synthesizing proinsulin. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3727–3731. doi: 10.1073/pnas.75.8.3727. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Weinberger J. Z., Greene M. I., Benacerraf B., Dorf M. E. Hapten-specific T-cell responses to 4-hydroxy-3-nitrophenyl acetyl. I. Genetic control of delayed-type hypersensitivity by VH and I-A-region genes. J Exp Med. 1979 Jun 1;149(6):1336–1348. doi: 10.1084/jem.149.6.1336. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wickens M. P., Buell G. N., Schimke R. T. Synthesis of double-stranded DNA complementary to lysozyme, ovomucoid, and ovalbumin mRNAs. Optimization for full length second strand synthesis by Escherichia coli DNA polymerase I. J Biol Chem. 1978 Apr 10;253(7):2483–2495. [PubMed] [Google Scholar]
- de Martynoff G., Pays E., Vassart G. Synthesis of a full length DNA complementary to thyroglobulin 33 S messenger RNA. Biochem Biophys Res Commun. 1980 Apr 14;93(3):645–653. doi: 10.1016/0006-291x(80)91127-4. [DOI] [PubMed] [Google Scholar]