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. 1991 Apr 15;88(8):3014–3018. doi: 10.1073/pnas.88.8.3014

Class II major histocompatibility complex molecules of murine dendritic cells: synthesis, sialylation of invariant chain, and antigen processing capacity are down-regulated upon culture.

E Kämpgen 1, N Koch 1, F Koch 1, P Stöger 1, C Heufler 1, G Schuler 1, N Romani 1
PMCID: PMC51374  PMID: 2014224

Abstract

Dendritic cells (DCs), such as Langerhans cells (LCs) of the epidermis and the DCs of lymphoid organs such as spleen, are potent antigen presenting cells. DCs express high levels of major histocompatibility complex (MHC) class II molecules, but, partly because of the low numbers of primary DCs in any tissue, there has been no detailed study of the biochemistry of their class II molecules. This information may be needed to help explain recent findings that DCs process native protein antigens when freshly isolated from epidermis and spleen. Processing ceases during culture, yet a strong accessory function for activating resting T cells develops. We studied immunoprecipitates of DC class II and invariant chain (Ii) molecules by two-dimensional gel electrophoresis. We found that (i) freshly isolated LCs synthesize large amounts of class II and Ii polypeptides; (ii) Ii molecules that are known to be involved in antigen processing display an unusually large number of sialic acids in fresh LCs; (iii) with culture, class II and Ii synthesis decreases dramatically and has virtually ceased at 3 days; and (iv) the turnover of class II in pulse/chase experiments is slow, being undetectable over a 12- to 32-hr culture period, whereas the turnover of Ii is rapid. We conclude that MHC class II molecules of DCs do not seem to be qualitatively unique. However, the regulation of class II and Ii expression is distinctive in that biosynthesis proceeds vigorously for a short period of time and the newly synthesized class II remains stably on the cell surface, whereas Ii turns over rapidly. This may enable DCs to process and retain antigens in the peripheral tissues such as skin and migrate to the lymphoid organs to activate T cells there.

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Selected References

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

  1. Blum J. S., Diaz R., Diment S., Fiani M., Mayorga L., Rodman J. S., Stahl P. D. Proteolytic processing in endosomal vesicles. Cold Spring Harb Symp Quant Biol. 1989;54(Pt 1):287–292. doi: 10.1101/sqb.1989.054.01.036. [DOI] [PubMed] [Google Scholar]
  2. Boog C. J., Neefjes J. J., Boes J., Ploegh H. L., Melief C. J. Specific immune responses restored by alteration in carbohydrate chains of surface molecules on antigen-presenting cells. Eur J Immunol. 1989 Mar;19(3):537–542. doi: 10.1002/eji.1830190319. [DOI] [PubMed] [Google Scholar]
  3. Brown J. H., Jardetzky 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. Nature. 1988 Apr 28;332(6167):845–850. doi: 10.1038/332845a0. [DOI] [PubMed] [Google Scholar]
  4. Crowley M., Inaba K., Steinman R. M. Dendritic cells are the principal cells in mouse spleen bearing immunogenic fragments of foreign proteins. J Exp Med. 1990 Jul 1;172(1):383–386. doi: 10.1084/jem.172.1.383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Crowley M., Inaba K., Witmer-Pack M., Steinman R. M. The cell surface of mouse dendritic cells: FACS analyses of dendritic cells from different tissues including thymus. Cell Immunol. 1989 Jan;118(1):108–125. doi: 10.1016/0008-8749(89)90361-4. [DOI] [PubMed] [Google Scholar]
  6. Davis J. E., Cresswell P. Lack of detectable endocytosis of B lymphocyte MHC class II antigens using an antibody-independent technique. J Immunol. 1990 Feb 1;144(3):990–997. [PubMed] [Google Scholar]
  7. Fischer H. G., Opel B., Reske K., Reske-Kunz A. B. Granulocyte-macrophage colony-stimulating factor-cultured bone marrow-derived macrophages reveal accessory cell function and synthesis of MHC class II determinants in the absence of external stimuli. Eur J Immunol. 1988 Aug;18(8):1151–1158. doi: 10.1002/eji.1830180802. [DOI] [PubMed] [Google Scholar]
  8. Guagliardi L. E., Koppelman B., Blum J. S., Marks M. S., Cresswell P., Brodsky F. M. Co-localization of molecules involved in antigen processing and presentation in an early endocytic compartment. Nature. 1990 Jan 11;343(6254):133–139. doi: 10.1038/343133a0. [DOI] [PubMed] [Google Scholar]
  9. Harding C. V., Roof R. W., Unanue E. R. Turnover of Ia-peptide complexes is facilitated in viable antigen-presenting cells: biosynthetic turnover of Ia vs. peptide exchange. Proc Natl Acad Sci U S A. 1989 Jun;86(11):4230–4234. doi: 10.1073/pnas.86.11.4230. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hirayama Y., Inaba K., Inaba M., Kato T., Kitaura M., Hosokawa T., Ikehara S., Muramatsu S. Neuraminidase-treated macrophages stimulate allogenic CD8+ T cells in the presence of exogenous interleukin 2. J Exp Med. 1988 Oct 1;168(4):1443–1456. doi: 10.1084/jem.168.4.1443. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Inaba K., Metlay J. P., Crowley M. T., Steinman R. M. Dendritic cells pulsed with protein antigens in vitro can prime antigen-specific, MHC-restricted T cells in situ. J Exp Med. 1990 Aug 1;172(2):631–640. doi: 10.1084/jem.172.2.631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Inaba K., Romani N., Steinman R. M. An antigen-independent contact mechanism as an early step in T cell-proliferative responses to dendritic cells. J Exp Med. 1989 Aug 1;170(2):527–542. doi: 10.1084/jem.170.2.527. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Inaba K., Schuler G., Witmer M. D., Valinksy J., Atassi B., Steinman R. M. Immunologic properties of purified epidermal Langerhans cells. Distinct requirements for stimulation of unprimed and sensitized T lymphocytes. J Exp Med. 1986 Aug 1;164(2):605–613. doi: 10.1084/jem.164.2.605. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Inaba K., Steinman R. M. Accessory cell-T lymphocyte interactions. Antigen-dependent and -independent clustering. J Exp Med. 1986 Feb 1;163(2):247–261. doi: 10.1084/jem.163.2.247. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Inaba K., Steinman R. M. Monoclonal antibodies to LFA-1 and to CD4 inhibit the mixed leukocyte reaction after the antigen-dependent clustering of dendritic cells and T lymphocytes. J Exp Med. 1987 May 1;165(5):1403–1417. doi: 10.1084/jem.165.5.1403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Koch F., Heufler C., Kämpgen E., Schneeweiss D., Böck G., Schuler G. Tumor necrosis factor alpha maintains the viability of murine epidermal Langerhans cells in culture, but in contrast to granulocyte/macrophage colony-stimulating factor, without inducing their functional maturation. J Exp Med. 1990 Jan 1;171(1):159–171. doi: 10.1084/jem.171.1.159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Koch N., Koch S., Hämmerling G. J. Ia invariant chain detected on lymphocyte surfaces by monoclonal antibody. Nature. 1982 Oct 14;299(5884):644–645. doi: 10.1038/299644a0. [DOI] [PubMed] [Google Scholar]
  18. Koch N., Lauer W., Habicht J., Dobberstein B. Primary structure of the gene for the murine Ia antigen-associated invariant chains (Ii). An alternatively spliced exon encodes a cysteine-rich domain highly homologous to a repetitive sequence of thyroglobulin. EMBO J. 1987 Jun;6(6):1677–1683. doi: 10.1002/j.1460-2075.1987.tb02417.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Koch N., Lipp J., Pessara U., Schenck K., Wraight C., Dobberstein B. MHC class II invariant chains in antigen processing and presentation. Trends Biochem Sci. 1989 Sep;14(9):383–386. doi: 10.1016/0968-0004(89)90013-3. [DOI] [PubMed] [Google Scholar]
  20. Koch N. Posttranslational modifications of the Ia-associated invariant protein p41 after gene transfer. Biochemistry. 1988 May 31;27(11):4097–4102. doi: 10.1021/bi00411a028. [DOI] [PubMed] [Google Scholar]
  21. Kraal G., Breel M., Janse M., Bruin G. Langerhans' cells, veiled cells, and interdigitating cells in the mouse recognized by a monoclonal antibody. J Exp Med. 1986 Apr 1;163(4):981–997. doi: 10.1084/jem.163.4.981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lakey E. K., Casten L. A., Niebling W. L., Margoliash E., Pierce S. K. Time dependence of B cell processing and presentation of peptide and native protein antigens. J Immunol. 1988 May 15;140(10):3309–3314. [PubMed] [Google Scholar]
  23. Larsen C. P., Steinman R. M., Witmer-Pack M., Hankins D. F., Morris P. J., Austyn J. M. Migration and maturation of Langerhans cells in skin transplants and explants. J Exp Med. 1990 Nov 1;172(5):1483–1493. doi: 10.1084/jem.172.5.1483. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lipp J., Dobberstein B. Signal recognition particle-dependent membrane insertion of mouse invariant chain: a membrane-spanning protein with a cytoplasmically exposed amino terminus. J Cell Biol. 1986 Jun;102(6):2169–2175. doi: 10.1083/jcb.102.6.2169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Macatonia S. E., Knight S. C., Edwards A. J., Griffiths S., Fryer P. Localization of antigen on lymph node dendritic cells after exposure to the contact sensitizer fluorescein isothiocyanate. Functional and morphological studies. J Exp Med. 1987 Dec 1;166(6):1654–1667. doi: 10.1084/jem.166.6.1654. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Machamer C. E., Cresswell P. Biosynthesis and glycosylation of the invariant chain associated with HLA-DR antigens. J Immunol. 1982 Dec;129(6):2564–2569. [PubMed] [Google Scholar]
  27. Marchalonis J. J., Cone R. E., Santer V. Enzymic iodination. A probe for accessible surface proteins of normal and neoplastic lymphocytes. Biochem J. 1971 Oct;124(5):921–927. doi: 10.1042/bj1240921. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. McCoy K. L., Miller J., Jenkins M., Ronchese F., Germain R. N., Schwartz R. H. Diminished antigen processing by endosomal acidification mutant antigen-presenting cells. J Immunol. 1989 Jul 1;143(1):29–38. [PubMed] [Google Scholar]
  29. Metlay J. P., Puré E., Steinman R. M. Control of the immune response at the level of antigen-presenting cells: a comparison of the function of dendritic cells and B lymphocytes. Adv Immunol. 1989;47:45–116. doi: 10.1016/s0065-2776(08)60662-8. [DOI] [PubMed] [Google Scholar]
  30. Metlay J. P., Witmer-Pack M. D., Agger R., Crowley M. T., Lawless D., Steinman R. M. The distinct leukocyte integrins of mouse spleen dendritic cells as identified with new hamster monoclonal antibodies. J Exp Med. 1990 May 1;171(5):1753–1771. doi: 10.1084/jem.171.5.1753. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Neefjes J. J., De Bruijn M. L., Boog C. J., Nieland J. D., Boes J., Melief C. J., Ploegh H. L. N-linked glycan modification on antigen-presenting cells restores an allospecific cytotoxic T cell response. J Exp Med. 1990 Feb 1;171(2):583–588. doi: 10.1084/jem.171.2.583. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Puré E., Inaba K., Crowley M. T., Tardelli L., Witmer-Pack M. D., Ruberti G., Fathman G., Steinman R. M. Antigen processing by epidermal Langerhans cells correlates with the level of biosynthesis of major histocompatibility complex class II molecules and expression of invariant chain. J Exp Med. 1990 Nov 1;172(5):1459–1469. doi: 10.1084/jem.172.5.1459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Roche P. A., Cresswell P. Invariant chain association with HLA-DR molecules inhibits immunogenic peptide binding. Nature. 1990 Jun 14;345(6276):615–618. doi: 10.1038/345615a0. [DOI] [PubMed] [Google Scholar]
  34. Romani N., Inaba K., Puré E., Crowley M., Witmer-Pack M., Steinman R. M. A small number of anti-CD3 molecules on dendritic cells stimulate DNA synthesis in mouse T lymphocytes. J Exp Med. 1989 Mar 1;169(3):1153–1168. doi: 10.1084/jem.169.3.1153. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Romani N., Koide S., Crowley M., Witmer-Pack M., Livingstone A. M., Fathman C. G., Inaba K., Steinman R. M. Presentation of exogenous protein antigens by dendritic cells to T cell clones. Intact protein is presented best by immature, epidermal Langerhans cells. J Exp Med. 1989 Mar 1;169(3):1169–1178. doi: 10.1084/jem.169.3.1169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Romani N., Schuler G. Structural and functional relationships between epidermal Langerhans cells and dendritic cells. Res Immunol. 1989 Nov-Dec;140(9):895–926. doi: 10.1016/0923-2494(89)90050-3. [DOI] [PubMed] [Google Scholar]
  37. Romani N., Stingl G., Tschachler E., Witmer M. D., Steinman R. M., Shevach E. M., Schuler G. The Thy-1-bearing cell of murine epidermis. A distinctive leukocyte perhaps related to natural killer cells. J Exp Med. 1985 Jun 1;161(6):1368–1383. doi: 10.1084/jem.161.6.1368. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Schuler G., Steinman R. M. Murine epidermal Langerhans cells mature into potent immunostimulatory dendritic cells in vitro. J Exp Med. 1985 Mar 1;161(3):526–546. doi: 10.1084/jem.161.3.526. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Shimada S., Caughman S. W., Sharrow S. O., Stephany D., Katz S. I. Enhanced antigen-presenting capacity of cultured Langerhans' cells is associated with markedly increased expression of Ia antigen. J Immunol. 1987 Oct 15;139(8):2551–2555. [PubMed] [Google Scholar]
  40. Steinman R. M., Nogueira N., Witmer M. D., Tydings J. D., Mellman I. S. Lymphokine enhances the expression and synthesis of Ia antigens on cultured mouse peritoneal macrophages. J Exp Med. 1980 Nov 1;152(5):1248–1261. doi: 10.1084/jem.152.5.1248. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Stockinger B., Pessara U., Lin R. H., Habicht J., Grez M., Koch N. A role of Ia-associated invariant chains in antigen processing and presentation. Cell. 1989 Feb 24;56(4):683–689. doi: 10.1016/0092-8674(89)90590-4. [DOI] [PubMed] [Google Scholar]
  42. Stössel H., Koch F., Kämpgen E., Stöger P., Lenz A., Heufler C., Romani N., Schuler G. Disappearance of certain acidic organelles (endosomes and Langerhans cell granules) accompanies loss of antigen processing capacity upon culture of epidermal Langerhans cells. J Exp Med. 1990 Nov 1;172(5):1471–1482. doi: 10.1084/jem.172.5.1471. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Witmer-Pack M. D., Valinsky J., Olivier W., Steinman R. M. Quantitation of surface antigens on cultured murine epidermal Langerhans cells: rapid and selective increase in the level of surface MHC products. J Invest Dermatol. 1988 Mar;90(3):387–394. doi: 10.1111/1523-1747.ep12456460. [DOI] [PubMed] [Google Scholar]
  44. Wolff K., Schreiner E. Uptake, intracellular transport and degradation of exogenous protein by Langerhans cells. An electron microscopic-cytochemical study using peroxidase as tracer substance. J Invest Dermatol. 1970 Jan;54(1):37–47. doi: 10.1111/1523-1747.ep12551501. [DOI] [PubMed] [Google Scholar]

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