Skip to main content
SpringerLink
Log in

Assembly, Intracellular Transport, and Release of MHC Class II Peptide Receptors

  • Protocol
  • First Online:
Antigen Processing

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1988))

Abstract

MHC class II molecules play a pivotal role for the induction and maintenance of immune responses against pathogens, but are also implicated in pathological conditions like autoimmune diseases or rejection of transplanted organs. Human antigen-presenting cells express three human leukocyte antigen (HLA) class II isotypes (DR, DP, and DQ), which are, with the exception of DRα, composed of highly polymorphic α and β subunits. The combination of α- and β-chains results in a multitude of MHC-II αβ-heterodimers of the same isotype, but also isotype-mixed MHC class II molecules have been identified. Invariant chain chaperones the assembly of MHC-II molecules within the endoplasmatic reticulum and also facilitates the intracellular transport to MHC class II loading compartments (MIICs). MHC-II molecules are loaded with antigenic peptides and shuttled to the cell surface for inspection by CD4 T-cells. Alternatively, class-II molecules enriched on intraluminal vesicles can be released via exosomes into the extracellular space. Since some of the αβ-combinations may yield mismatched nonfunctional heterodimers, it is not entirely clear which type of HLA class II peptide receptors are transported to MIICs and found on the cell surface of antigen-presenting cells. We present techniques to inspect assembly, intracellular transport, cell surface expression, and exosomal release of MHC class II heterodimers.

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

Access this chapter

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Similar content being viewed by others

References

  1. Ayala García MA, González Yebra B, López Flores AL, Guaní Guerra E (2012) The major histocompatibility complex in transplantation. J Transp Secur 2012:842141

    Google Scholar 

  2. Piertney SB, Oliver MK (2005) The evolutionary ecology of the major histocompatibility complex. Heredity 96:7

    Article  Google Scholar 

  3. Schooten E, Di Maggio A, van Bergen En Henegouwen PMP, Kijanka MM (2018) MAGE-A antigens as targets for cancer immunotherapy. Cancer Treat Rev 67:54–62

    Article  CAS  Google Scholar 

  4. You L, Mao L, Wei J, Jin S, Yang C, Liu H, Zhu L, Qian W (2017) The crosstalk between autophagic and endo-/exosomal pathways in antigen processing for MHC presentation in anticancer T cell immune responses. J Hematol Oncol 10:165

    Article  Google Scholar 

  5. Ermann J, Fathman CG (2001) Autoimmune diseases: genes, bugs and failed regulation. Nat Immunol 2:759

    Article  CAS  Google Scholar 

  6. Blander JM (2018) Regulation of the cell biology of antigen cross-presentation. Annu Rev Immunol 36:717–753

    Article  CAS  Google Scholar 

  7. Valečka J, Almeida CR, Su B, Pierre P, Gatti E (2018) Autophagy and MHC-restricted antigen presentation. Mol Immunol 99:163–170

    Article  Google Scholar 

  8. Robinson J, Halliwell JA, Hayhurst JD, Flicek P, Parham P, Marsh SG (2014) The IPD and IMGT/HLA database: allele variant databases. Nucleic Acids Res 43:D423–D431

    Article  Google Scholar 

  9. Gotoh Y, Takashima H, Noguchi K, Nishimura H, Tokushima M, Shirai T, Kimoto M (1993) Mixed haplotype A beta Z/A alpha d class II molecule in (NZB x NZW)F1 mice detected by T cell clones. J Immunol 150:4777–4787

    CAS  PubMed  Google Scholar 

  10. Kimoto M (1996) Mixed-haplotype and mixed-isotype major histocompatibility complex class II molecules detected by murine T-cell clones and their possible involvement in autoimmunity. Methods 9:430–438

    Article  CAS  Google Scholar 

  11. Koeleman BP, Lie BA, Undlien DE, Dudbridge F, Thorsby E, De Vries RR, Cucca F, Roep BO, Giphart M, Todd JA (2004) Genotype effects and epistasis in type 1 diabetes and HLA-DQ trans dimer associations with disease. Genes Immun 5:381

    Article  CAS  Google Scholar 

  12. Malissen B, Shastri N, Pierres M, Hood L (1986) Cotransfer of the Ed alpha and Ad beta genes into L cells results in the surface expression of a functional mixed-isotype Ia molecule. Proc Natl Acad Sci U S A 83:3958–3962

    Article  CAS  Google Scholar 

  13. Silk JD, Schoendorf D, Bartok I, Chai J-G, Gray D, Simpson E, Dyson J (2005) Mixed-haplotype MHC class II molecules select functional CD4+ T cells. Mol Immunol 42:1129–1139

    Article  CAS  Google Scholar 

  14. Germain RN, Quill H (1986) Unexpected expression of a unique mixed-isotype class II MHC molecule by transfected L-cells. Nature 320:72–75

    Article  CAS  Google Scholar 

  15. Lotteau V, Teyton L, Burroughs D, Charron D (1987) A novel HLA class II molecule (DR alpha-DQ beta) created by mismatched isotype pairing. Nature 329:339–341

    Article  CAS  Google Scholar 

  16. Ruberti G, Sellins KS, Hill CM, Germain RN, Fathman CG, Livingstone A (1992) Presentation of antigen by mixed isotype class II molecules in normal H-2d mice. J Exp Med 175:157–162

    Article  CAS  Google Scholar 

  17. Spencer JS, Kubo RT (1989) Mixed isotype class II antigen expression. A novel class II molecule is expressed on a murine B cell lymphoma. J Exp Med 169:625–640

    Article  CAS  Google Scholar 

  18. Temme S, Zacharias M, Neumann J, Wohlfromm S, König A, Temme N, Springer S, Trowsdale J, Koch N (2014) A novel family of human leukocyte antigen class II receptors may have its origin in archaic human species. J Biol Chem 289:639–653

    Article  CAS  Google Scholar 

  19. Koch N, Moldenhauer G, Hofmann W, Möller P (1991) Rapid intracellular pathway gives rise to cell surface expression of the MHC class II-associated invariant chain (CD74). J Immunol 147:2643–2651

    CAS  PubMed  Google Scholar 

  20. McCormick PJ, Martina JA, Bonifacino JS (2005) Involvement of clathrin and AP-2 in the trafficking of MHC class II molecules to antigen-processing compartments. Proc Natl Acad Sci 102:7910–7915

    Article  CAS  Google Scholar 

  21. Garstka MA, Neefjes J (2013) How to target MHC class II into the MIIC compartment. Mol Immunol 55:162–165

    Article  CAS  Google Scholar 

  22. Sadegh-Nasseri S, Chen M, Narayan K, Bouvier M (2008) The convergent roles of tapasin and HLA-DM in antigen presentation. Trends Immunol 29:141–147

    Article  CAS  Google Scholar 

  23. Wubbolts R, Neefjes J (1999) Intracellular transport and peptide loading of MHC class II molecules: regulation by chaperones and motors. Immunol Rev 172:189–208

    Article  CAS  Google Scholar 

  24. Babbitt BP, Allen PM, Matsueda G, Haber E, Unanue ER (1985) Binding of immunogenic peptides to Ia histocompatibility molecules. Nature 317:359–361

    Article  CAS  Google Scholar 

  25. Meldolesi J (2018) Exosomes and ectosomes in intercellular communication. Curr Biol 28:R435–R444. https://doi.org/10.1016/j.cub.2018.01.059

    Article  CAS  PubMed  Google Scholar 

  26. Poe AJ, Knowlton AA (2018) Exosomes and cardiovascular cell–cell communication. Essays Biochem 62:193–204. https://doi.org/10.1042/EBC20170081

    Article  PubMed  Google Scholar 

  27. Théry C, Duban L, Segura E, Véron P, Lantz O, Amigorena S (2002) Indirect activation of naïve CD4+ T cells by dendritic cell-derived exosomes. Nat Immunol 3:1156–1162

    Article  Google Scholar 

  28. Temme S, Eis-Hübinger AM, McLellan AD, Koch N (2010) The herpes simplex virus-1 encoded glycoprotein B diverts HLA-DR into the exosome pathway. J Immunol 184:236–243

    Article  CAS  Google Scholar 

  29. Shackelford DA, Strominger J (1983) Analysis of the oligosaccharides on the HLA-DR and DC1 B cell antigens. J Immunol 130:274–282

    CAS  PubMed  Google Scholar 

  30. Germain RN, Hendrix LR (1991) MHC class II structure, occupancy and surface expression determined by post-endoplasmic reticulum antigen binding. Nature 353:134–139

    Article  CAS  Google Scholar 

  31. Mellins E, Smith L, Arp B, Cotner T, Celis E, Pious D (1990) Defective processing and presentation of exogenous antigens in mutants with normal HLA class II genes. Nature 343:71–74

    Article  CAS  Google Scholar 

  32. Watanabe M, Suzuki T, Taniguchi M, Shinohara N (1983) Monoclonal anti-Ia murine alloantibodies crossreactive with the Ia-homologues of other mammalian species including humans. Transplantation 36:712–718

    Article  CAS  Google Scholar 

  33. Pesando JM, Graf L (1986) Differential expression of HLA-DR, -DQ, and -DP antigens on malignant B cells. J Immunol 136:4311–4318

    CAS  PubMed  Google Scholar 

  34. Temponi M, Kekish U, Hamby CV, Nielsen H, Marboe CC, Ferrone S (1993) Characterization of anti-HLA class II monoclonal antibody LGII-612.14 reacting with formalin fixed tissues. J Immunol Methods 161:239–256

    Article  CAS  Google Scholar 

  35. Lampson LA, Levy R (1980) Two populations of Ia-like molecules on a human B cell line. J Immunol 125:293–299

    CAS  PubMed  Google Scholar 

  36. Hitzel C, Grüneberg U, van Ham M, Trowsdale J, Koch N (1999) Sodium dodecyl sulfate-resistant HLA-DR “superdimer” bands are in some cases class II heterodimers bound to antibody. J Immunol 162:4671–4676

    CAS  PubMed  Google Scholar 

  37. Accolla RS, Gross N, Carrel S, Corte G (1981) Distinct forms of both alpha and beta subunits are present in the human Ia molecular pool. Proc Natl Acad Sci U S A 78:4549–4551

    Article  CAS  Google Scholar 

  38. Charron D, McDevitt H (1980) Characterization of HLA-D-region antigens by two-dimensional gel electrophoresis. Molecular-genotyping. J Exp Med 152:18s–36s

    CAS  PubMed  Google Scholar 

  39. Adams T, Bodmer J, Bodmer W (1983) Production and characterization of monoclonal antibodies recognizing the alpha-chain subunits of human ia alloantigens. Immunology 50:613

    CAS  PubMed  PubMed Central  Google Scholar 

  40. Ting YT, Temme S, Koch N, McLellan AD (2009) A new monoclonal antibody recognizing a linear determinant on the HLA-DRα chain N-terminus. Hybridoma 28:423–429

    Article  CAS  Google Scholar 

  41. Spits H, Borst J, Giphart M, Coligan J, Terhorst C, de Vries JE (1984) HLA-DC antigens can serve as recognition elements for human cytotoxic T lymphocytes. Eur J Immunol 14:299–304

    Article  CAS  Google Scholar 

  42. Shookster L, Matsuyama T, Burmester G, Winchester R (1987) Monoclonal antibody 1a3 recognizes a monomorphic epitope unique to DQ molecules. Hum Immunol 20:59–70

    Article  CAS  Google Scholar 

  43. Wraight CJ, Van Endert P, Möller P, Lipp J, Ling N, MacLennan I, Koch N, Moldenhauer G (1990) Human major histocompatibility complex class II invariant chain is expressed on the cell surface. J Biol Chem 265:5787–5792

    CAS  PubMed  Google Scholar 

  44. Denzin LK, Robbins NF, Carboy-Newcomb C, Cresswell P (1994) Assembly and intracellular transport of HLA-DM and correction of the class II antigen-processing defect in T2 cells. Immunity 1:595–606

    Article  CAS  Google Scholar 

  45. Southern JA, Young D, Heaney F, Baumgärtner W, Randall R (1991) Identification of an epitope on the P and V proteins of simian virus 5 that distinguishes between two isolates with different biological characteristics. J Gen Virol 72:1551–1557

    Article  CAS  Google Scholar 

  46. Kurth D, Neumann J, Demleitner K, Hildmann J, Mehlig M, Scheicher C, Wendling U, Sun D, Reske K (1997) Full length cDNA of rat RT1. DMa and RT1. DMb and expression of RT1. DM genes in dendritic and Langerhans cells. Biol Chem 378:1005–1012

    Article  CAS  Google Scholar 

  47. Neumann J (2005) Novel antibody tags from the rat lysosomal protein RT1. DM for immunodetection of recombinant proteins. J Immunol Methods 301:66–76

    Article  CAS  Google Scholar 

  48. Paul P, van den Hoorn T, Jongsma MLM, Bakker MJ, Hengeveld R, Janssen L, Cresswell P, Egan DA, van Ham M, ten Brinke A, Ovaa H, Beijersbergen RL, Kuijl C, Neefjes J (2011) A genome-wide multidimensional RNAi screen reveals pathways controlling MHC class II antigen presentation. Cell 145:268–283

    Article  CAS  Google Scholar 

  49. Schindelin J, Arganda-Carreras I, Frise E, Kaynig V, Longair M, Pietzsch T, Preibisch S, Rueden C, Saalfeld S, Schmid B, Tinevez J-Y, White DJ, Hartenstein V, Eliceiri K, Tomancak P, Cardona A (2012) Fiji: an open-source platform for biological-image analysis. Nat Methods 9:676–682

    Article  CAS  Google Scholar 

  50. Koch N (1988) Posttranslational modifications of the Ia-associated invariant protein p41 after gene transfer. Biochemistry (Mosc) 27:4097–4102

    Article  CAS  Google Scholar 

  51. Kämper N, Franken S, Temme S, Koch S, Bieber T, Koch N (2012) γ-Interferon-regulated chaperone governs human lymphocyte antigen class II expression. FASEB J 26:104–116

    Article  Google Scholar 

  52. Johnson JP, Demmer-Dieckmann M, Meo T, Hadam MR, Riethmüller G (1981) Surface antigens of human melanoma cells defined by monoclonal antibodies. I. Biochemical characterization of two antigens found on cell lines and fresh tumors of diverse tissue origin. Eur J Immunol 11:825–831

    Article  CAS  Google Scholar 

  53. Koch N, Zacharias M, König A, Temme S, Neumann J, Springer S (2011) Stoichiometry of HLA class II-invariant chain oligomers. PLoS One 6:e17257

    Article  CAS  Google Scholar 

  54. Pearson H (2007) The good, the bad and the ugly. Nature 447:138–140

    Article  CAS  Google Scholar 

  55. Bolte S, Cordelières FP (2006) A guided tour into subcellular colocalization analysis in light microscopy. J Microsc 224:213–232

    Article  CAS  Google Scholar 

  56. Niazy N*, Temme S*, Bocuk D, Giesen C, Konig A, Temme N, Ziegfeld A, Gregers TF, Bakke O, Lang T, Eis-Hübinger AM, Koch N (2017) Misdirection of endosomal trafficking mediated by herpes simplex virus-encoded glycoprotein B. FASEB J 31:1650–1667

    Article  CAS  Google Scholar 

Download references

Acknowledgments

We thank Angelika König for expert technical assistance.

Author information

Authors and Affiliations

  1. Department of Molecular Cardiology, Heinrich-Heine-University of Düsseldorf, Düsseldorf, Germany

    Sebastian Temme

  2. Division of Immunobiology, Institute of Genetics, University of Bonn, Bonn, Germany

    Nadine Temme & Norbert Koch

Authors
  1. Sebastian Temme
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Nadine Temme
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Norbert Koch
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Sebastian Temme .

Editor information

Editors and Affiliations

  1. INSERM U1151, Université Paris Descartes, Hopital Necker-Enfants Malades, Paris, France

    Peter van Endert

Rights and permissions

Reprints and permissions

Copyright information

© 2019 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Temme, S., Temme, N., Koch, N. (2019). Assembly, Intracellular Transport, and Release of MHC Class II Peptide Receptors. In: van Endert, P. (eds) Antigen Processing. Methods in Molecular Biology, vol 1988. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9450-2_22

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-9450-2_22

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9449-6

  • Online ISBN: 978-1-4939-9450-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation