Skip to main content
NCBI home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1995 Feb 1;181(2):493–502. doi: 10.1084/jem.181.2.493

Cells expressing a major histocompatibility complex class I molecule with a single covalently bound peptide are highly immunogenic

PMCID: PMC2191874  PMID: 7836906

Abstract

The major histocompatibility complex (MHC) class I molecules expressed at the cell surface are associated with a large number of different peptides so that the density of a given MHC-peptide complex is relatively low. Here we describe the properties of MHC class I molecules genetically attached to a single antigenic peptide. Cells expressing these fusion proteins are recognized by T cells specific for the particular MHC-peptide complex. Coculture of naive splenocytes with cells expressing these MHC-peptide fusion proteins and the B7.1 antigen allows the induction of primary cytotoxic T lymphocytes (CTL) in vitro. Injection of these cells into naive mice enhances the frequency of specific CTL precursors and protects against a subsequent challenge with a tumor or a virus bearing the antigenic peptide. Soluble MHC- peptide fusions were also produced in which all three components, that is, the heavy chain, beta 2-microglobulin and the peptide, have fused into a single-chain protein. The availability of MHC class I molecules bound to a single peptide provides valuable tools for the manipulation of CTL responses and the analysis of the selection processes in the thymus.

Full Text

The Full Text of this article is available as a PDF (1.0 MB).

Selected References

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

  1. Abastado J. P., Jaulin C., Schutze M. P., Langlade-Demoyen P., Plata F., Ozato K., Kourilsky P. Fine mapping of epitopes by intradomain Kd/Dd recombinants. J Exp Med. 1987 Aug 1;166(2):327–340. doi: 10.1084/jem.166.2.327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Abastado J. P., Ojcius D. M., Casrouge A., Yeh P., Schumacher T. N., Ploegh H. L., Kourilsky P. A soluble, single-chain Kd molecule produced by yeast selects a peptide repertoire indistinguishable from that of cell-surface-associated Kd. Eur J Immunol. 1993 Aug;23(8):1776–1783. doi: 10.1002/eji.1830230807. [DOI] [PubMed] [Google Scholar]
  3. Bednarek M. A., Sauma S. Y., Gammon M. C., Porter G., Tamhankar S., Williamson A. R., Zweerink H. J. The minimum peptide epitope from the influenza virus matrix protein. Extra and intracellular loading of HLA-A2. J Immunol. 1991 Dec 15;147(12):4047–4053. [PubMed] [Google Scholar]
  4. Bellio M., Lone Y. C., de la Calle-Martin O., Malissen B., Abastado J. P., Kourilsky P. The V beta complementarity determining region 1 of a major histocompatibility complex (MHC) class I-restricted T cell receptor is involved in the recognition of peptide/MHC I and superantigen/MHC II complex. J Exp Med. 1994 Apr 1;179(4):1087–1097. doi: 10.1084/jem.179.4.1087. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bjorkman P. J., Saper M. A., Samraoui B., Bennett W. S., Strominger J. L., Wiley D. C. Structure of the human class I histocompatibility antigen, HLA-A2. Nature. 1987 Oct 8;329(6139):506–512. doi: 10.1038/329506a0. [DOI] [PubMed] [Google Scholar]
  6. Casanova J. L., Cerottini J. C., Matthes M., Necker A., Gournier H., Barra C., Widmann C., MacDonald H. R., Lemonnier F., Malissen B. H-2-restricted cytolytic T lymphocytes specific for HLA display T cell receptors of limited diversity. J Exp Med. 1992 Aug 1;176(2):439–447. doi: 10.1084/jem.176.2.439. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Casanova J. L., Martinon F., Gournier H., Barra C., Pannetier C., Regnault A., Kourilsky P., Cerottini J. C., Maryanski J. L. T cell receptor selection by and recognition of two class I major histocompatibility complex-restricted antigenic peptides that differ at a single position. J Exp Med. 1993 Mar 1;177(3):811–820. doi: 10.1084/jem.177.3.811. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chen Y., Sidney J., Southwood S., Cox A. L., Sakaguchi K., Henderson R. A., Appella E., Hunt D. F., Sette A., Engelhard V. H. Naturally processed peptides longer than nine amino acid residues bind to the class I MHC molecule HLA-A2.1 with high affinity and in different conformations. J Immunol. 1994 Mar 15;152(6):2874–2881. [PubMed] [Google Scholar]
  9. Cox A. L., Skipper J., Chen Y., Henderson R. A., Darrow T. L., Shabanowitz J., Engelhard V. H., Hunt D. F., Slingluff C. L., Jr Identification of a peptide recognized by five melanoma-specific human cytotoxic T cell lines. Science. 1994 Apr 29;264(5159):716–719. doi: 10.1126/science.7513441. [DOI] [PubMed] [Google Scholar]
  10. De Bruijn M. L., Nieland J. D., Schumacher T. N., Ploegh H. L., Kast W. M., Melief C. J. Mechanisms of induction of primary virus-specific cytotoxic T lymphocyte responses. Eur J Immunol. 1992 Nov;22(11):3013–3020. doi: 10.1002/eji.1830221137. [DOI] [PubMed] [Google Scholar]
  11. De Bruijn M. L., Schumacher T. N., Nieland J. D., Ploegh H. L., Kast W. M., Melief C. J. Peptide loading of empty major histocompatibility complex molecules on RMA-S cells allows the induction of primary cytotoxic T lymphocyte responses. Eur J Immunol. 1991 Dec;21(12):2963–2970. doi: 10.1002/eji.1830211210. [DOI] [PubMed] [Google Scholar]
  12. Fremont D. H., Matsumura M., Stura E. A., Peterson P. A., Wilson I. A. Crystal structures of two viral peptides in complex with murine MHC class I H-2Kb. Science. 1992 Aug 14;257(5072):919–927. doi: 10.1126/science.1323877. [DOI] [PubMed] [Google Scholar]
  13. Garrett T. P., Saper M. A., Bjorkman P. J., Strominger J. L., Wiley D. C. Specificity pockets for the side chains of peptide antigens in HLA-Aw68. Nature. 1989 Dec 7;342(6250):692–696. doi: 10.1038/342692a0. [DOI] [PubMed] [Google Scholar]
  14. Godeau F., Luescher I. F., Ojcius D. M., Saucier C., Mottez E., Cabanie L., Kourilsky P. Purification and ligand binding of a soluble class I major histocompatibility complex molecule consisting of the first three domains of H-2Kd fused to beta 2-microglobulin expressed in the baculovirus-insect cell system. J Biol Chem. 1992 Dec 5;267(34):24223–24229. [PubMed] [Google Scholar]
  15. Guo H. C., Jardetzky T. S., Garrett T. P., Lane W. S., Strominger J. L., Wiley D. C. Different length peptides bind to HLA-Aw68 similarly at their ends but bulge out in the middle. Nature. 1992 Nov 26;360(6402):364–366. doi: 10.1038/360364a0. [DOI] [PubMed] [Google Scholar]
  16. Harding C. V., Unanue E. R. Quantitation of antigen-presenting cell MHC class II/peptide complexes necessary for T-cell stimulation. Nature. 1990 Aug 9;346(6284):574–576. doi: 10.1038/346574a0. [DOI] [PubMed] [Google Scholar]
  17. Houbiers J. G., Nijman H. W., van der Burg S. H., Drijfhout J. W., Kenemans P., van de Velde C. J., Brand A., Momburg F., Kast W. M., Melief C. J. In vitro induction of human cytotoxic T lymphocyte responses against peptides of mutant and wild-type p53. Eur J Immunol. 1993 Sep;23(9):2072–2077. doi: 10.1002/eji.1830230905. [DOI] [PubMed] [Google Scholar]
  18. Huston J. S., Levinson D., Mudgett-Hunter M., Tai M. S., Novotný J., Margolies M. N., Ridge R. J., Bruccoleri R. E., Haber E., Crea R. Protein engineering of antibody binding sites: recovery of specific activity in an anti-digoxin single-chain Fv analogue produced in Escherichia coli. Proc Natl Acad Sci U S A. 1988 Aug;85(16):5879–5883. doi: 10.1073/pnas.85.16.5879. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Jaulin C., Romero P., Luescher I. F., Casanova J. L., Prochnicka-Chalufour A., Langlade-Demoyen P., Maryanski J. L., Kourilsky P. Most residues on the floor of the antigen binding site of the class I MHC molecule H-2Kd influence peptide presentation. Int Immunol. 1992 Aug;4(8):945–953. doi: 10.1093/intimm/4.8.945. [DOI] [PubMed] [Google Scholar]
  20. Kahn-Perles B., Barra C., Jehan J., Fourel D., Barad M., Maryanski J. L., Lemonnier F. A. Cytotoxic T lymphocyte recognition of secreted HLA class I molecules. J Immunol. 1989 May 1;142(9):3021–3025. [PubMed] [Google Scholar]
  21. Kourilsky P., Claverie J. M. The peptidic self model: a hypothesis on the molecular nature of the immunological self. Ann Inst Pasteur Immunol. 1986 Jul-Aug;137D(1):3–21. [PubMed] [Google Scholar]
  22. Kozono H., White J., Clements J., Marrack P., Kappler J. Production of soluble MHC class II proteins with covalently bound single peptides. Nature. 1994 May 12;369(6476):151–154. doi: 10.1038/369151a0. [DOI] [PubMed] [Google Scholar]
  23. Lalanne J. L., Transy C., Guerin S., Darche S., Meulien P., Kourilsky P. Expression of class I genes in the major histocompatibility complex: identification of eight distinct mRNAs in DBA/2 mouse liver. Cell. 1985 Jun;41(2):469–478. doi: 10.1016/s0092-8674(85)80020-9. [DOI] [PubMed] [Google Scholar]
  24. Langlade-Demoyen P., Levraud J. P., Kourilsky P., Abastado J. P. Primary cytotoxic T lymphocyte induction using peptide-stripped autologous cells. Int Immunol. 1994 Nov;6(11):1759–1766. doi: 10.1093/intimm/6.11.1759. [DOI] [PubMed] [Google Scholar]
  25. Letourneur F., Malissen B. Derivation of a T cell hybridoma variant deprived of functional T cell receptor alpha and beta chain transcripts reveals a nonfunctional alpha-mRNA of BW5147 origin. Eur J Immunol. 1989 Dec;19(12):2269–2274. doi: 10.1002/eji.1830191214. [DOI] [PubMed] [Google Scholar]
  26. Madden D. R., Garboczi D. N., Wiley D. C. The antigenic identity of peptide-MHC complexes: a comparison of the conformations of five viral peptides presented by HLA-A2. Cell. 1993 Nov 19;75(4):693–708. doi: 10.1016/0092-8674(93)90490-h. [DOI] [PubMed] [Google Scholar]
  27. Madden D. R., Gorga J. C., Strominger J. L., Wiley D. C. The structure of HLA-B27 reveals nonamer self-peptides bound in an extended conformation. Nature. 1991 Sep 26;353(6342):321–325. doi: 10.1038/353321a0. [DOI] [PubMed] [Google Scholar]
  28. Madden D. R., Gorga J. C., Strominger J. L., Wiley D. C. The three-dimensional structure of HLA-B27 at 2.1 A resolution suggests a general mechanism for tight peptide binding to MHC. Cell. 1992 Sep 18;70(6):1035–1048. doi: 10.1016/0092-8674(92)90252-8. [DOI] [PubMed] [Google Scholar]
  29. Mage M. G., Lee L., Ribaudo R. K., Corr M., Kozlowski S., McHugh L., Margulies D. H. A recombinant, soluble, single-chain class I major histocompatibility complex molecule with biological activity. Proc Natl Acad Sci U S A. 1992 Nov 15;89(22):10658–10662. doi: 10.1073/pnas.89.22.10658. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Maryanski J. L., Accolla R. S., Jordan B. H2-restricted recognition of cloned HLA class I gene products expressed in mouse cells. J Immunol. 1986 Jun 15;136(12):4340–4347. [PubMed] [Google Scholar]
  31. Maryanski J. L., Romero P., Van Pel A., Boon T., Salemme F. R., Cerottini J. C., Corradin G. The identification of tyrosine as a common key residue in unrelated H-2Kd restricted antigenic peptides. Int Immunol. 1991 Oct;3(10):1035–1042. doi: 10.1093/intimm/3.10.1035. [DOI] [PubMed] [Google Scholar]
  32. Matsumura M., Fremont D. H., Peterson P. A., Wilson I. A. Emerging principles for the recognition of peptide antigens by MHC class I molecules. Science. 1992 Aug 14;257(5072):927–934. doi: 10.1126/science.1323878. [DOI] [PubMed] [Google Scholar]
  33. Miettinen H. M., Rose J. K., Mellman I. Fc receptor isoforms exhibit distinct abilities for coated pit localization as a result of cytoplasmic domain heterogeneity. Cell. 1989 Jul 28;58(2):317–327. doi: 10.1016/0092-8674(89)90846-5. [DOI] [PubMed] [Google Scholar]
  34. Mottez E., Jaulin C., Godeau F., Choppin J., Levy J. P., Kourilsky P. A single-chain murine class I major transplantation antigen. Eur J Immunol. 1991 Feb;21(2):467–471. doi: 10.1002/eji.1830210232. [DOI] [PubMed] [Google Scholar]
  35. 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]
  36. Ojcius D. M., Langlade-Demoyen P., Gachelin G., Kourilsky P. Role for MHC class I molecules in selecting and protecting high affinity peptides in the presence of proteases. J Immunol. 1994 Mar 15;152(6):2798–2810. [PubMed] [Google Scholar]
  37. Pala P., Corradin G., Strachan T., Sodoyer R., Jordan B. R., Cerottini J. C., Maryanski J. L. Mapping of HLA epitopes recognized by H-2-restricted cytotoxic T lymphocytes specific for HLA using recombinant genes and synthetic peptides. J Immunol. 1988 Feb 1;140(3):871–877. [PubMed] [Google Scholar]
  38. Rötzschke O., Falk K., Deres K., Schild H., Norda M., Metzger J., Jung G., Rammensee H. G. Isolation and analysis of naturally processed viral peptides as recognized by cytotoxic T cells. Nature. 1990 Nov 15;348(6298):252–254. doi: 10.1038/348252a0. [DOI] [PubMed] [Google Scholar]
  39. Schutze M. P., Langlade-Demoyen P., Przewlocki G., Leclerc C. Alloantigen pretreatment induces a down-regulation of the in vivo subsequent development of cytotoxic T lymphocytes directed against linked alloantigens. Eur J Immunol. 1989 Aug;19(8):1365–1371. doi: 10.1002/eji.1830190803. [DOI] [PubMed] [Google Scholar]
  40. Silver M. L., Guo H. C., Strominger J. L., Wiley D. C. Atomic structure of a human MHC molecule presenting an influenza virus peptide. Nature. 1992 Nov 26;360(6402):367–369. doi: 10.1038/360367a0. [DOI] [PubMed] [Google Scholar]
  41. Townsend A., Bodmer H. Antigen recognition by class I-restricted T lymphocytes. Annu Rev Immunol. 1989;7:601–624. doi: 10.1146/annurev.iy.07.040189.003125. [DOI] [PubMed] [Google Scholar]
  42. Trucco M., de Petris S., Garotta G., Ceppellini R. Quantitative analysis of cell surface HLA structures by means of monoclonal antibodies. Hum Immunol. 1980 Oct;1(3):233–243. doi: 10.1016/0198-8859(80)90018-x. [DOI] [PubMed] [Google Scholar]
  43. Urban R. G., Chicz R. M., Lane W. S., Strominger J. L., Rehm A., Kenter M. J., UytdeHaag F. G., Ploegh H., Uchanska-Ziegler B., Ziegler A. A subset of HLA-B27 molecules contains peptides much longer than nonamers. Proc Natl Acad Sci U S A. 1994 Feb 15;91(4):1534–1538. doi: 10.1073/pnas.91.4.1534. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Zhang W., Young A. C., Imarai M., Nathenson S. G., Sacchettini J. C. Crystal structure of the major histocompatibility complex class I H-2Kb molecule containing a single viral peptide: implications for peptide binding and T-cell receptor recognition. Proc Natl Acad Sci U S A. 1992 Sep 1;89(17):8403–8407. doi: 10.1073/pnas.89.17.8403. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES