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. 1996 Jul 9;93(14):7184–7189. doi: 10.1073/pnas.93.14.7184

Covalent assembly of a soluble T cell receptor-peptide-major histocompatibility class I complex.

C Grégoire 1, S Y Lin 1, G Mazza 1, N Rebai 1, I F Luescher 1, B Malissen 1
PMCID: PMC38957  PMID: 8692966

Abstract

We used stepwise photochemical cross-linking for specifically assembling soluble and covalent complexes made of a T-cell antigen receptor (TCR) and a class I molecule of the major histocompatibility complex (MHC) bound to an antigenic peptide. For that purpose, we have produced in myeloma cells a single-chain Fv construct of a TCR specific for a photoreactive H-2Kd-peptide complex. Photochemical cross-linking of this TCR single-chain Fv with a soluble form of the photoreactive H-2Kd-peptide ligand resulted in the formation of a ternary covalent complex. We have characterized the soluble ternary complex and showed that it reacted with antibodies specific for epitopes located either on the native TCR or on the Kd molecules. By preventing the fast dissociation kinetics observed with most T cell receptors, this approach provides a means of preparing soluble TCR-peptide-MHC complexes on large-scale levels.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abastado J. P., Lone Y. C., Casrouge A., Boulot G., Kourilsky P. Dimerization of soluble major histocompatibility complex-peptide complexes is sufficient for activation of T cell hybridoma and induction of unresponsiveness. J Exp Med. 1995 Aug 1;182(2):439–447. doi: 10.1084/jem.182.2.439. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bentley G. A., Boulot G., Karjalainen K., Mariuzza R. A. Crystal structure of the beta chain of a T cell antigen receptor. Science. 1995 Mar 31;267(5206):1984–1987. doi: 10.1126/science.7701320. [DOI] [PubMed] [Google Scholar]
  3. Carbone A. M., Marrack P., Kappler J. W. Remethylation at sites 5' of the murine Lyt-2 gene in association with shutdown of Lyt-2 expression. J Immunol. 1988 Aug 15;141(4):1369–1375. [PubMed] [Google Scholar]
  4. Chang H. C., Bao Z., Yao Y., Tse A. G., Goyarts E. C., Madsen M., Kawasaki E., Brauer P. P., Sacchettini J. C., Nathenson S. G. A general method for facilitating heterodimeric pairing between two proteins: application to expression of alpha and beta T-cell receptor extracellular segments. Proc Natl Acad Sci U S A. 1994 Nov 22;91(24):11408–11412. doi: 10.1073/pnas.91.24.11408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chien Y. H., Davis M. M. How alpha beta T-cell receptors 'see' peptide/MHC complexes. Immunol Today. 1993 Dec;14(12):597–602. doi: 10.1016/0167-5699(93)90199-u. [DOI] [PubMed] [Google Scholar]
  6. Chung S., Wucherpfennig K. W., Friedman S. M., Hafler D. A., Strominger J. L. Functional three-domain single-chain T-cell receptors. Proc Natl Acad Sci U S A. 1994 Dec 20;91(26):12654–12658. doi: 10.1073/pnas.91.26.12654. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Corr M., Slanetz A. E., Boyd L. F., Jelonek M. T., Khilko S., al-Ramadi B. K., Kim Y. S., Maher S. E., Bothwell A. L., Margulies D. H. T cell receptor-MHC class I peptide interactions: affinity, kinetics, and specificity. Science. 1994 Aug 12;265(5174):946–949. doi: 10.1126/science.8052850. [DOI] [PubMed] [Google Scholar]
  8. Engel I., Ottenhoff T. H., Klausner R. D. High-efficiency expression and solubilization of functional T cell antigen receptor heterodimers. Science. 1992 May 29;256(5061):1318–1321. doi: 10.1126/science.1598575. [DOI] [PubMed] [Google Scholar]
  9. Grégoire C., Rebaï N., Schweisguth F., Necker A., Mazza G., Auphan N., Millward A., Schmitt-Verhulst A. M., Malissen B. Engineered secreted T-cell receptor alpha beta heterodimers. Proc Natl Acad Sci U S A. 1991 Sep 15;88(18):8077–8081. doi: 10.1073/pnas.88.18.8077. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gunning P., Leavitt J., Muscat G., Ng S. Y., Kedes L. A human beta-actin expression vector system directs high-level accumulation of antisense transcripts. Proc Natl Acad Sci U S A. 1987 Jul;84(14):4831–4835. doi: 10.1073/pnas.84.14.4831. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hilyard K. L., Reyburn H., Chung S., Bell J. I., Strominger J. L. Binding of soluble natural ligands to a soluble human T-cell receptor fragment produced in Escherichia coli. Proc Natl Acad Sci U S A. 1994 Sep 13;91(19):9057–9061. doi: 10.1073/pnas.91.19.9057. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hoo W. F., Lacy M. J., Denzin L. K., Voss E. W., Jr, Hardman K. D., Kranz D. M. Characterization of a single-chain T-cell receptor expressed in Escherichia coli. Proc Natl Acad Sci U S A. 1992 May 15;89(10):4759–4763. doi: 10.1073/pnas.89.10.4759. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kappler J., White J., Kozono H., Clements J., Marrack P. Binding of a soluble alpha beta T-cell receptor to superantigen/major histocompatibility complex ligands. Proc Natl Acad Sci U S A. 1994 Aug 30;91(18):8462–8466. doi: 10.1073/pnas.91.18.8462. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kubo R. T., Born W., Kappler J. W., Marrack P., Pigeon M. Characterization of a monoclonal antibody which detects all murine alpha beta T cell receptors. J Immunol. 1989 Apr 15;142(8):2736–2742. [PubMed] [Google Scholar]
  15. Kurucz I., Jost C. R., George A. J., Andrew S. M., Segal D. M. A bacterially expressed single-chain Fv construct from the 2B4 T-cell receptor. Proc Natl Acad Sci U S A. 1993 May 1;90(9):3830–3834. doi: 10.1073/pnas.90.9.3830. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Luescher I. F., Anjuère F., Peitsch M. C., Jongeneel C. V., Cerottini J. C., Romero P. Structural analysis of TCR-ligand interactions studied on H-2Kd-restricted cloned CTL specific for a photoreactive peptide derivative. Immunity. 1995 Jul;3(1):51–63. doi: 10.1016/1074-7613(95)90158-2. [DOI] [PubMed] [Google Scholar]
  17. Luescher I. F., Cerottini J. C., Romero P. Photoaffinity labeling of the T cell receptor on cloned cytotoxic T lymphocytes by covalent photoreactive ligand. J Biol Chem. 1994 Feb 25;269(8):5574–5582. [PubMed] [Google Scholar]
  18. Luescher I. F., Vivier E., Layer A., Mahiou J., Godeau F., Malissen B., Romero P. CD8 modulation of T-cell antigen receptor-ligand interactions on living cytotoxic T lymphocytes. Nature. 1995 Jan 26;373(6512):353–356. doi: 10.1038/373353a0. [DOI] [PubMed] [Google Scholar]
  19. Madden D. R. The three-dimensional structure of peptide-MHC complexes. Annu Rev Immunol. 1995;13:587–622. doi: 10.1146/annurev.iy.13.040195.003103. [DOI] [PubMed] [Google Scholar]
  20. Matsui K., Boniface J. J., Reay P. A., Schild H., Fazekas de St Groth B., Davis M. M. Low affinity interaction of peptide-MHC complexes with T cell receptors. Science. 1991 Dec 20;254(5039):1788–1791. doi: 10.1126/science.1763329. [DOI] [PubMed] [Google Scholar]
  21. Matsui K., Boniface J. J., Steffner P., Reay P. A., Davis M. M. Kinetics of T-cell receptor binding to peptide/I-Ek complexes: correlation of the dissociation rate with T-cell responsiveness. Proc Natl Acad Sci U S A. 1994 Dec 20;91(26):12862–12866. doi: 10.1073/pnas.91.26.12862. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Matsumura M., Saito Y., Jackson M. R., Song E. S., Peterson P. A. In vitro peptide binding to soluble empty class I major histocompatibility complex molecules isolated from transfected Drosophila melanogaster cells. J Biol Chem. 1992 Nov 25;267(33):23589–23595. [PubMed] [Google Scholar]
  23. Novotny J., Ganju R. K., Smiley S. T., Hussey R. E., Luther M. A., Recny M. A., Siliciano R. F., Reinherz E. L. A soluble, single-chain T-cell receptor fragment endowed with antigen-combining properties. Proc Natl Acad Sci U S A. 1991 Oct 1;88(19):8646–8650. doi: 10.1073/pnas.88.19.8646. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Pantoliano M. W., Bird R. E., Johnson S., Asel E. D., Dodd S. W., Wood J. F., Hardman K. D. Conformational stability, folding, and ligand-binding affinity of single-chain Fv immunoglobulin fragments expressed in Escherichia coli. Biochemistry. 1991 Oct 22;30(42):10117–10125. doi: 10.1021/bi00106a007. [DOI] [PubMed] [Google Scholar]
  25. Reiter Y., Kurucz I., Brinkmann U., Jung S. H., Lee B., Segal D. M., Pastan I. Construction of a functional disulfide-stabilized TCR Fv indicates that antibody and TCR Fv frameworks are very similar in structure. Immunity. 1995 Mar;2(3):281–287. doi: 10.1016/1074-7613(95)90052-7. [DOI] [PubMed] [Google Scholar]
  26. Seth A., Stern L. J., Ottenhoff T. H., Engel I., Owen M. J., Lamb J. R., Klausner R. D., Wiley D. C. Binary and ternary complexes between T-cell receptor, class II MHC and superantigen in vitro. Nature. 1994 May 26;369(6478):324–327. doi: 10.1038/369324a0. [DOI] [PubMed] [Google Scholar]
  27. Skerra A. Bacterial expression of immunoglobulin fragments. Curr Opin Immunol. 1993 Apr;5(2):256–262. doi: 10.1016/0952-7915(93)90014-j. [DOI] [PubMed] [Google Scholar]
  28. Strong R. K., Penny D. M., Feldman R. M., Weiner L. P., Boniface J. J., Davis M. M., Bjorkman P. J. Engineering and expression of a secreted murine TCR with reduced N-linked glycosylation. J Immunol. 1994 Nov 1;153(9):4111–4121. [PubMed] [Google Scholar]
  29. Sykulev Y., Brunmark A., Jackson M., Cohen R. J., Peterson P. A., Eisen H. N. Kinetics and affinity of reactions between an antigen-specific T cell receptor and peptide-MHC complexes. Immunity. 1994 Apr;1(1):15–22. doi: 10.1016/1074-7613(94)90005-1. [DOI] [PubMed] [Google Scholar]
  30. Sykulev Y., Brunmark A., Tsomides T. J., Kageyama S., Jackson M., Peterson P. A., Eisen H. N. High-affinity reactions between antigen-specific T-cell receptors and peptides associated with allogeneic and syngeneic major histocompatibility complex class I proteins. Proc Natl Acad Sci U S A. 1994 Nov 22;91(24):11487–11491. doi: 10.1073/pnas.91.24.11487. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Weber S., Traunecker A., Oliveri F., Gerhard W., Karjalainen K. Specific low-affinity recognition of major histocompatibility complex plus peptide by soluble T-cell receptor. Nature. 1992 Apr 30;356(6372):793–796. doi: 10.1038/356793a0. [DOI] [PubMed] [Google Scholar]
  32. White J., Blackman M., Bill J., Kappler J., Marrack P., Gold D. P., Born W. Two better cell lines for making hybridomas expressing specific T cell receptors. J Immunol. 1989 Sep 15;143(6):1822–1825. [PubMed] [Google Scholar]

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