Skip to main content

Advertisement

SpringerLink
Log in

Analysis of antigen presentation using HLA transfectants

  • MHC Class II Genes and Molecules, Symposium (Part I)
  • Published:
Immunologic Research Aims and scope Submit manuscript

Conclusions

In summary, the transfection of HLA class II sequences is proving a useful tool for various aspects of the analysis of antigen presentation. Such transfectants have been used in the generation of new polymorphic HLA mAbs and in mapping the specificities of existing mAbs. In terms of analysing the HLA restriction of particular T cell clones, transfectant APC are increasingly being used as a means of identifying the presenting class II molecule, a less equivocal approach than mAb blocking studies or presentation by B-LCL panels. Our studies with DR2-restricted clones showed that, as with other haplotypes which generate two DR products, one of the two expressed DRβ chains is functionally dominant. Once restricting molecules for interesting clones have been worked out, spe cific sites in the cDNA can be readily mutagenised to establish the areas of interaction between peptide, class II and TCR. On the more general question of APC function, transfected L cells are capable not only of presenting peptide antigens but also process complex antigens, suggesting that some of the degradative pathways necessary for processing are generally available in diverse cell types. The ability of transfectants to function as APC seems to extend, at least in some cases, to the presentation of allodeterminants. HLA class II transfected L cells which were poor at presenting either soluble antigen or allo-antigen to T cells owing to insufficient levels of class II expression could be made into potent APC by retransfection with ICAM-1, restoring the accessory molecule interaction between LFA-1 and ICAM-1. This approach is proving valuable in defining the role of such accessory molecules in antigen presentation. With the ability to generate powerful tools for the in vitro dissection of HLA function as well as the appearance of transgenic mice for in vivo studies, the coming years should yield exciting advances in understanding the complexities of antigen presentation in human immunity.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Shimonkevitz, R.J.; Kappler, J.W.; Marrack, P.; Grey, H.: Antigen recognition by H-2-restricted T cells. I. Cell-free antigen processing. J. exp. Med.158: 303–316 (1983).

    Article  PubMed  CAS  Google Scholar 

  2. Babbitt, B.P.; Allen, P.M.; Matsueda, G.; Haber, E.; Unanue, E.R.: Binding of immunogenic peptides to Ia histocompatibility molecules. Nature317: 359–361 (1985).

    Article  PubMed  CAS  Google Scholar 

  3. Bell, J.I.; Denney, D.; Foster, L.; Belt, T.; Todd, J.A.; McDevitt, H.O.: Allelic variation in the DR subregion of the human major histocompatibility complex. Proc natn. Acad. Sci. USA84: 6234–6238 (1987).

    Article  CAS  Google Scholar 

  4. Todd, J.A.; Bell, J.I.; McDevitt, H.O.: HLA-DQβ gene contributes to susceptibility and resistance to insulin-dependent diabetes mellitus. Nature329: 599–604 (1987).

    Article  PubMed  CAS  Google Scholar 

  5. Bugawan, T.L.; Horn, G.T.; Long, C.M.; Mickelson, E.; Hansen, J.A.; Ferrara, G.B.; Angelini, G.; Erlich, H.A.: Analysis of HLA-DP allelic sequence polymorphism using the in vitro enzymatic DNA amplification of DPα and DPβ loci. J. Immun.141: 4024–4031 (1988).

    PubMed  CAS  Google Scholar 

  6. Bjorkmann, P.J.; Saper, M.A.; Samraoui, B.; Bennett, W.S.; Strominger, J.L.; Wiley, D.C.: The foreign antigen binding site. Nature329: 512–518 (1988).

    Article  Google Scholar 

  7. Brown, J.H.; Jardetsky, T.; Saper, M.A.; Samraoui, B.; Bjorkman, P.J.; Wiley, D.C.: A hypothetical model of the foreign antigen binding site of class II histocompatibility molecules. Nature332: 845–847 (1988).

    Article  PubMed  CAS  Google Scholar 

  8. Davis, M.M.; Bjorkmann, P.J.: T cell antigen receptor genes and T cell recognition. Nature334: 395–401 (1988).

    Article  PubMed  CAS  Google Scholar 

  9. Austin, P.; Trowsdale, J.; Rudd, C.; Bodmer, W.; Feldmann, M.; Lamb, J.: Functional expression of HLA-DP genes transfected into mouse fibroblasts. Nature13: 61–64 (1985).

    Article  Google Scholar 

  10. Wilkinson, D.; Altmann, D.M.; Ikeda, H.; Trowsdale, J.: Analysis of HLA-class II products by DNA-mediated gene transfer (in press).

  11. Wilkinson, D.; de Vries, R.R.P.; Madrigal, J.A.; Lock, C.B.; Trowsdale, J.; Altmann, D.M.: Analysis of HLA-DR glycoproteins by DNA-mediated gene transfer. Definition of DR2β gene products and antigen presentation to T cell clones from leprosy patients. J. exp. Med.167: 1442–1458 (1988).

    Article  PubMed  CAS  Google Scholar 

  12. Ikeda, H.; Trowsdale, J.; Saito, I.: Mulcos: a vector for amplification and simultaneous expression of two foreign genes in mammalian cells. Gene71: 19–27 (1988).

    Article  PubMed  CAS  Google Scholar 

  13. Korman, A.J.; Frantz, J.D.; Strominger, J.L.; Mulligan, R.C.: Expression of human class II histocompatibility antigens using a retrovirus vector. Proc. natn. Acad. Sci. USA82: 2150–2153 (1987).

    Article  Google Scholar 

  14. Kwok, W.W.; Schwarz, D.; Nepom, B.S.; Hock, R.A.; Thurtle, P.S.; Nepom, G.T.: HLA-DQ molecules form α-β heterodimers of mixed allotype. J. Immun.141: 3123–3127 (1989).

    Google Scholar 

  15. Lotteau, V.; Sands, J.; Teyton, L.; Turmel, P.; Charron, D.; Strominger, J.L.: Modulation of HLA class II antigen expression by transfection of sense and antisense DRα cDNA chain. J. exp. Med.169: 351–356 (1989).

    Article  PubMed  CAS  Google Scholar 

  16. Heyes, J.; Austin, P.J.; Bodmer, J.; Bodmer, W.F.; Madrigal, A.; Mazzilli, M.; Trowsdale, J.: Monoclonal antibodies to HLA-DP transfected mouse L cells. Proc natn. Acad. Sci. USA83: 3417–3421 (1986).

    Article  CAS  Google Scholar 

  17. Young, J.A.T.; Lindsay, J.; Bodmer, J.G.; Trowsdale, J.: Epitope recognition by a DPα chain specific monoclonal anbitody (DP11.1) is influenced by the interaction between the DPα chain and its polymorphic DPβ chain partner. Hum. Immunol.23: 37–44 (1988).

    Article  PubMed  CAS  Google Scholar 

  18. Klohe, E.P.; Watts, R.; Bahl, M.; Alber, C.; Yu, W.-Y.; Anderson, R.; Silver, J.; Gregerson, P.K.; Karr, R.W.: Analysis of molecular specificities of anti-class II monoclonal antibodies by using L cell transfectants expressing HLA class II molecules. J. Immun.141: 2158–2164 (1988).

    PubMed  CAS  Google Scholar 

  19. Eckels, D.D.; Woody, J.N.; Hartzman, R.J.: Monoclonal and xenoantibodies inhibit primary responses to HLA-DR but fail to inhibit secondary proliferative (PLT) responses to allogeneic cells. Hum. Immunol.3: 133–142 (1981).

    Article  PubMed  CAS  Google Scholar 

  20. Young, J.A.T.; Wilkinson, D.; Bodmer, W.F.; Trowsdale, J.: Sequence and evolution of HLA-DR7 and DRw53 associated β chains. Proc natn. Acad. Sci.84: 4929–4933 (1987).

    Article  CAS  Google Scholar 

  21. Ozato, K.; Mayer, N.; Sachs, D.H.: Hybridoma cell lines secreting monoclonal antibodies to mouse H-2 and I-A antigens. J. Immun.124: 533–549 (1980).

    PubMed  CAS  Google Scholar 

  22. Ziegler, A.B.; Uchanska-Ziegler, J.; Zeuthen, J.; Wernet, P.: HLA-antigen expression at the single cell level on a K562x B cell hybrid: an analysis with monoclonal antibodies using bacterial binding assays. Somatic Cell Genet.8: 775–789 (1982).

    Article  PubMed  CAS  Google Scholar 

  23. Lampson, L.A.; Levy, R.: Two populations of Ia-like molecules on a human B cell line. J. Immun.125: 293–299 (1980).

    PubMed  CAS  Google Scholar 

  24. Rothbard, J.B.; Lechler, R.I.; Howland, K.; Bal, V.; Eckels, D.; Sekaly, R.; Long, E.O.; Taylor, W.R.; Lamb, J.R.: Structural model of HLA-DR1 restricted T cell antigen recognition. Cell52: 515–523 (1988).

    Article  PubMed  CAS  Google Scholar 

  25. Siliciano, R.F.; Lawton, T.; Knall, C.; Karr, R.W.; Berman, P.; Gregory, T.; Reinherz, E.L.: Host virus interaction in AIDS: analysis with human T cell clones reveals complexities of T cell receptor recognition and suggests a mechanism for CD4+ cell depletion. Cell54: 561–575 (1988).

    Article  PubMed  CAS  Google Scholar 

  26. Lamb, J.R.; Rees, A.D.M.; Bal, V.; Ikeda, H.; Wilkinson, D.; de Vries, R.R.P.; Rothbard, J.B.: Prediction and identification of an HLA-DR-restricted T cell determinant in the 19kDa protein ofM. tuberculosis. Eur. J. Immunol.18: 973–976 (1988).

    Article  PubMed  CAS  Google Scholar 

  27. Celis, E.; Karr, R.W.: Presentation of an immunodominant T cell epitope of hepatitis B surface antigen by the HLA-DPw4 molecule. J. Virol.63: 747–755 (1989).

    PubMed  CAS  Google Scholar 

  28. Sekaly, R.P.; Jacobson, S.; Richert, J.R.; Tonnelle, C.; McFarland, H.F.; Long, E.O.: Antigen presentation to HLA class II-restricted measles virus-specific T cell clones can occur in the absence of the invariant chain. Proc natn. Acad. Sci. USA85: 1209–1212 (1988).

    Article  CAS  Google Scholar 

  29. Eckels D.D.; Sell, T.W.; Long, E.O.; Sekaly, R.P.: Presentation of influenza hemagglutinin peptide in the presence of limited allostimulation by HLA-DR1 transfected human fibroblasts. Hum. Immunol.21: 173–181 (1988).

    Article  PubMed  CAS  Google Scholar 

  30. Shastri, N.; Malissen, B.; Hood, L.: Ia-transfected L cell fibroblasts present a lysozyme peptide but not the native protein to lysozyme-specific T cells. Proc. natn. Acad. Sci. USA82: 5885–5889 (1985).

    Article  CAS  Google Scholar 

  31. Collins, T.; Korman, A.J.; Wake, C.T.; Boss, J.M.; Kappes, D.J.; Fiers, W.; Ault, K.A.; Gimbrone, M.A.; Strominger, J.L.; Pober, J.S.: Immune interferon activates multiple class II major histocompatibility complex genes and the associated invariant chain gene in human endothelial cells and the associated invariant chain gene in human endothelial cells and dermal fibroblasts. Proc. natn. Acad. Sci. USA81: 4917–4921 (1984).

    Article  CAS  Google Scholar 

  32. Wagner, C.R.; Vetto, R.M.; Burger, D.R.: The mechanism of antigen presentation by endothelial cells. Immunobiology168: 453–469 (1984).

    PubMed  CAS  Google Scholar 

  33. Botazzo, G.F.; Pujol-Borrell, R.; Hanafusa, T.; Feldmann, M.: Role of aberrant HLA-DR expression and antigen presentation by endothelial cells. Lancetii: 1115–1119 (1983).

    Article  Google Scholar 

  34. Londei, M.; Lamb, J.R.; Bottazzo, G.F.; Feldmann, M.: Epithelial cells expressing aberrant MHC class II determinants can present antigen to cloned human T cells. Nature312: 639–641 (1984).

    Article  PubMed  CAS  Google Scholar 

  35. Markmann, J.; Lo, D.; Naji, A.; Palmiter, R.L.; Brinster, R.L.; Heber-Katz, E.: Antigen-presenting function of class II MHC expressing pancreatic beta cells. Nature336: 476–479 (1988).

    Article  PubMed  CAS  Google Scholar 

  36. Buus, S.; Sette, A.; Grey, H.M.: The interaction between protein-derived immunogenic peptides and Ia. Immunol. Rev.98: 115–141 (1987).

    Article  PubMed  CAS  Google Scholar 

  37. Matzinger, P.; Bevan, M.J.: Why do so many lymphocytes respond to major histocompatibility antigens? Cell. Immunol.29: 1–5 (1977).

    Article  PubMed  CAS  Google Scholar 

  38. Marrack, P.; Kappler, J.: T cells can distinguish between allogeneic major histocompatibility complex products on different cell types. Nature332: 840–843 (1988).

    Article  PubMed  CAS  Google Scholar 

  39. Cowan, E.P.; Coligan, J.E.; Biddison, W.E.: Human cytotoxic T lymphocyte recognition of an HLA-A3 gene product expressed on murine L cells: the only human gene product required on the target cell for lysis is the class I heavy chain. Proc. natn. Acad. Sci USA82: 4490–4494 (1985).

    Article  CAS  Google Scholar 

  40. Barbosa, J.A.; Santos-Aguado, J.; Menzer, S.J.; Strominger, J.L.; Burakoff, S.J.; Biro, P.A.: Site directed mutagenesis of class I HLA genes: role of glycosylation in surface expression and functional recognition. Proc. natn. Acad. Sci. USA81: 7549–7553 (1984).

    Article  CAS  Google Scholar 

  41. Bernhard, E.J.; Le, A.-X.T.; Yannelli, J.R.; Holterman, M.J.: Hogan, K.T.; Parham, P.; Engelhard, V.H.: The ability of cytotoxic T cells to recognize HLA-A2.1 or HLA-B7 antigens expressed on murine cells correlates with their epitope specificity. J. Immun.139: 3614–3621 (1987).

    PubMed  CAS  Google Scholar 

  42. Nakatsuji, T.; Inoko, H.; Ando, A.; Sato, T.; Koide, T.; Tadakuma, T.; Yoshida, T.O.; Tsuji, K.: The role of transfected HLA-DQ genes in the mixed lymphocyte reaction-like condition. Immunogenetics25: 1–6 (1987).

    Article  PubMed  CAS  Google Scholar 

  43. Buss, S.; Sette, A.; Colon, S.M.; Grey, H.M.: Autologous peptides constitutively occupy the antigen-binding site on Ia. Science242: 1045–1047 (1988).

    Article  Google Scholar 

  44. Matis, L.A.; Sorger, S.B.; McElligot, D.L.; Fink, P.J.: Hedrick, S.M.: The molecular basis of alloreactivity in antigen-specific, major histocompatibility complex restricted T cell clones. Cell51: 59–69 (1987).

    Article  PubMed  CAS  Google Scholar 

  45. Marlin, S.; Springer, T.A.: Purified intercellular adhesion molecule-1 (ICAM-1) is a ligand for lymphocyte function-associated antigen-1 (LFA-1). Cell51: 813–819 (1987).

    Article  PubMed  CAS  Google Scholar 

  46. Simmons, D.; Makgoba, M.W.; Seed, B.: ICAM, an adhesion ligand of LFA-1 is homologous to the neural cell adhesion molecule NCAM. Nature331: 624–627 (1988).

    Article  PubMed  CAS  Google Scholar 

  47. Springer, T.A.; Dustin, M.L.; Kishimoto, T.K.; Marlin, S.D.: The lymphocyte function associated LFA-1, CD2 and LFA-3 molecules: cell adhesion receptors of the immune system. Annu. Rev. Immunol.5: 223–252 (1987).

    Article  PubMed  CAS  Google Scholar 

  48. Gregory, C.D.; Murray, R.J.; Edwards, C.F.; Rickinson, A.B.: Downregulation of cell adhesion molecules LFA-3 and ICAM-1 in Epstein-Barr-virus-positive Burkitt’s lymphoma underlies tumor cell escape from virus-specific T cell surveillance. J. exp. Med.167: 1811–1824 (1988).

    Article  PubMed  CAS  Google Scholar 

  49. Barbosa, J.A.; Mentzer, S.J.; Kamarck, M.E.; Hart, J.; Biro, P.A.; Strominger, J.L.; Burakoff, S.J.: Gene mapping and somatic cell hybrid analysis of the role of human lymphocyte function-associated antigen-3 (LFA-3) in CTL-target cell interactions. J. Immun.136: 3085–3091 (1986).

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Human Imunogenetics Laboratory, Imperial Cancer Research Fund, 44 Lincoln’s Inn Fields, WC2A 3PX, London, UK

    Daniel M. Altmann, David Wilkinson, Hitoshi Ikeda & John Trowsdale

Authors
  1. Daniel M. Altmann
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. David Wilkinson
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hitoshi Ikeda
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. John Trowsdale
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Altmann, D.M., Wilkinson, D., Ikeda, H. et al. Analysis of antigen presentation using HLA transfectants. Immunol Res 9, 57–68 (1990). https://doi.org/10.1007/BF02918479

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02918479

Keywords

Search

Navigation