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Quantitation of Major Histocompatibility Complex Antigen Endocytosis Using Capture ELISA

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Antigen Processing and Presentation Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 156))

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Abstract

Major histocompatibility class I and II (MHC-I and MHC-II) antigens (Ags), as well as a variety of cell surface receptors, have been shown to enter the endocytic pathway of antigen-presenting cells (APCs). MHC Ags endocytose into clathrin-coated pits, which pinch off from the plasma membrane and form clathrin-coated vesicles (1-4). These vesicles rapidly uncoat and fuse together or with pre-existing vesicular structures called “early endosomes.” Selected molecules within early endosomes return to the plasma membrane, possibly via carrier vesicles (5). This pathway has been best described for endocytosed transferrin receptors (TfR) (6).

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References

  1. Tse, D. B. and Pernis, B. (1984) Spontaneous internalization of class I major histocompatibility complex molecules in T lymphoid cells. J.Exp. Med. 159, 193–207.

    Article  CAS  PubMed  Google Scholar 

  2. Reid, P. A. and Watts, C. (1990) Cycling of cell surface MHC glycoproteins through primaquine-sensitive intracellular compartments. Nature 346, 655–657.

    Article  CAS  PubMed  Google Scholar 

  3. Machy, P., Truneh, A., Gennaro, D., and Hoffstein, S. (1987) Major histocompatibility complex class I molecules internalized via coated pits in T lymphocytes. Nature 328, 724–726.

    Article  CAS  PubMed  Google Scholar 

  4. Gruenberg, J. and Howell, K. E. (1989) Membrane traffic in endocytosis: insights from cell-free assays. Annu. Rev. Cell Biol. 5, 453–481.

    Article  CAS  PubMed  Google Scholar 

  5. Ajioka, R. S. and Kaplan, J. (1986) Intracellular pools of transferrin receptors result from constitutive internalization of unoccupied receptors. Proc. Natl. Acad. Sci. USA 83, 6445–6449.

    Article  CAS  PubMed  Google Scholar 

  6. Machy, P., Barbet, J., and Leserman, L. D. (1982) Differential endocytosis of T and B lymphocyte surface molecules evaluated with antibody-bearing fluorescent lipo-somes containing methotrexate. Proc. Natl. Acad. Sci. USA 79, 4148–4152.

    Article  CAS  PubMed  Google Scholar 

  7. Pernis, B. (1985) Internalization of lymphocyte membrane components. Immunol. Today6, 45–49.

    Google Scholar 

  8. Aragnol, D., Malissen, B., Schiff, C., Piron, M., and Leserman, L. D. (1986) Endocytosis of MHC molecules by B cell-B lymphoma and B cell-T lymphoma hybrids. J. Immunol. 137, 3347–3353.

    CAS  PubMed  Google Scholar 

  9. Motal, A., Zhou, X., Joke, A., Siddiqi, A. R., Srinivasa, B. R., Stenvall, K., Dahmen, J., and Jondal, M. (1993) Major histocompatibility complex class I-binding peptides are recycled to the cell surface after internalization. Eur. J. Immunol. 23, 3224–3229.

    Article  PubMed  Google Scholar 

  10. Anderson, M. S., Swier, K., Arneson, L., and Miller, J. (1993) Enhanced antigen presentation in the absence of the invariant chain endosomal localization signal. J. Exp. Med. 178, 1959–1969.

    Article  CAS  PubMed  Google Scholar 

  11. Peterson, M. and Miller, J. (1990) Invariant chain influences the immunological recognition of MHC class II molecules. Nature 345, 172–174.

    Article  CAS  PubMed  Google Scholar 

  12. Germain, R. N. and Rinker, A. G., Jr. (1993) Peptide binding inhibits protein aggregation of invariant-chain free class II dimers and promotes surface expression of occupied molecules. Nature 363, 725–728.

    Article  CAS  PubMed  Google Scholar 

  13. Chicz, R. M., Urban, R. G., Lane, W. S., Gorga, J. C., Stern, L. T., Vignali, D. A., and Strominger, J. L. (1992) Predominant naturally processed peptides bound to HLA-DR1 are derived from MHC-related molecules and are heterogeneous in size. Nature 358, 764–768.

    Article  CAS  PubMed  Google Scholar 

  14. Bretscher, M. S. and Lutter, R. (1988) A new method for detecting endocytosed proteins. EMBO J. 7, 4087–4092.

    CAS  PubMed  Google Scholar 

  15. Pinet, V., Vergelli, M., Martin, R., Bakke, O., and Long, E. O. (1995) Antigen presentation mediated by recycling of surface HLA-DR molecules. Nature 375, 603–606.

    Article  CAS  PubMed  Google Scholar 

  16. Brachet, V., Raposo, G., Amigorena, S., and Mellman, I. (1997) Invariant chain controls the transport of major histocompatibility complex class II molecules to and from lysosomes. J. Cell Biol. 137, 51–65.

    Article  CAS  PubMed  Google Scholar 

  17. Marmorstein, A. D., Bonilha, V. L., Chiflet, S., Neill, J. M., and Rodriguez-Boulan, E. (1996) The polarity of the plasma membrace protein RET-PE2 in retinal pigment epithelium is developmentally regulated. J. Cell Sci. 109, 3025–3034.

    CAS  PubMed  Google Scholar 

  18. Volz, B., Orberger, G., Porwoll, S., Hauri, H. P., and Tauber, R. (1995). Selective reentry of recycling cell surface glycoproteins to the biosynthetic pathway in human hepatocarcinoma HepG2 cells. J. Cell Biol. 130, 537–551.

    Article  CAS  PubMed  Google Scholar 

  19. Anderson, G. W., Zimmerman, J., and Callahan, F. (1964) The uses of esters of N-hydroxysuccinimide in peptide synthesis. J. Am. Chem. Soc. 86, 1839–1844.

    Article  CAS  Google Scholar 

  20. Amigorena, S., Webster, P., Drake, J., Newcomb, J., Cresswell, P., and Mellman, I. (1995) Invariant chain cleavage and peptide loading in major histocompatibility complex class II vesicles. J. Exp. Med. 181, 1729–1741.

    Article  CAS  PubMed  Google Scholar 

  21. Gallager, S., Winston, S. E., Fuller, S. A., and Hurrell, J. G. R. (1998) Immunoblotting and Immunodetection, in Current Protocols in Immunology (Coligan, J., Kruisbeek, A., Margulies, D., Shevach, E., and Strober, W., eds.), John Wiley, New York pp. 8.10.1–8.10.11.

    Google Scholar 

  22. Schwartz, A. L., Bolognesi, A., and Fridovich, S. E. (1984) Recycling of the asialoglycoprotein receptor and the effect of lysosomotropic amines in hepatoma cells. J. Cell Biol. 98, 732–738.

    Article  CAS  PubMed  Google Scholar 

  23. Stoorvogel, W., Oorschot, V., and Geuzem, J. (1996) A novel class of clatherin-coated vesicles budding from endosomes. J. Cell Biol. 132, 21–33.

    Article  CAS  PubMed  Google Scholar 

  24. Daukas, G. and Zigmond, S. H. (1985) Inhibition of receptor-mediated but not fluid-phase endocytosis in polymorphonuclear leukocytes. J. Cell Biol. 101, 1673–1679.

    Article  CAS  PubMed  Google Scholar 

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© 2000 Humana Press Inc.

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Turvy, D.N., Blum, J.S. (2000). Quantitation of Major Histocompatibility Complex Antigen Endocytosis Using Capture ELISA. In: Solheim, J.C. (eds) Antigen Processing and Presentation Protocols. Methods in Molecular Biology, vol 156. Humana Press. https://doi.org/10.1385/1-59259-062-4:57

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  • DOI: https://doi.org/10.1385/1-59259-062-4:57

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-745-8

  • Online ISBN: 978-1-59259-062-9

  • eBook Packages: Springer Protocols

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