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. 1997 Jul 1;100(1):204–215. doi: 10.1172/JCI119514

Intestinal epithelial cells use two distinct pathways for HLA class II antigen processing.

R M Hershberg 1, P E Framson 1, D H Cho 1, L Y Lee 1, S Kovats 1, J Beitz 1, J S Blum 1, G T Nepom 1
PMCID: PMC508181  PMID: 9202073

Abstract

Intestinal epithelial cells express a low level of HLA class II molecules constitutively, with elevated levels seen in the setting of mucosal inflammation including inflammatory bowel disease. The ability of intestinal epithelial cells to act as antigen presenting cells for alphabeta CD4(+) T lymphocytes was examined through a molecular analysis of the HLA class II antigen processing pathway. We have shown that intestinal epithelial cells contain abundant constitutive levels of the cathepsin proteases proven to function in HLA class II mediated antigen presentation. Activation of these cells by gamma-IFN induced the expression of invariant chain and HLA-DM alphabeta, thus facilitating the formation of compact, SDS-stable HLA- DR alphabeta heterodimers. Using HLA-DR-restricted T cells and retroviral mediated gene transfer of HLA-DR alleles into the intestinal epithelial cell lines HT-29 and T84, we demonstrated efficient antigen processing and presentation to CD4(+) T lymphocytes in the presence of the proinflammatory cytokine gamma-IFN. The class II processing pathway and presentation in the presence of gamma-IFN was indistinguishable from that observed with a conventional antigen presenting cell. Antigen processing also occurred in intestinal epithelial cells in the absence of gamma-IFN, and in contrast to that seen after stimulation with gamma-IFN, required high concentration of antigen and was not inhibited by the protease inhibitor leupeptin. These data suggest the use of two distinct pathways of HLA class II antigen processing in enterocytes with differential immunomodulatory properties in the presence or absence of mucosal inflammation.

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Selected References

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  1. Bakke O., Dobberstein B. MHC class II-associated invariant chain contains a sorting signal for endosomal compartments. Cell. 1990 Nov 16;63(4):707–716. doi: 10.1016/0092-8674(90)90137-4. [DOI] [PubMed] [Google Scholar]
  2. Barone K. S., Jain S. L., Michael J. G. Effect of in vivo depletion of CD4+ and CD8+ cells on the induction and maintenance of oral tolerance. Cell Immunol. 1995 Jun;163(1):19–29. doi: 10.1006/cimm.1995.1094. [DOI] [PubMed] [Google Scholar]
  3. Barrett A. J. Fluorimetric assays for cathepsin B and cathepsin H with methylcoumarylamide substrates. Biochem J. 1980 Jun 1;187(3):909–912. doi: 10.1042/bj1870909. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Barrett A. J., Kirschke H. Cathepsin B, Cathepsin H, and cathepsin L. Methods Enzymol. 1981;80(Pt 100):535–561. doi: 10.1016/s0076-6879(81)80043-2. [DOI] [PubMed] [Google Scholar]
  5. Bland P. W., Kambarage D. M. Antigen handling by the epithelium and lamina propria macrophages. Gastroenterol Clin North Am. 1991 Sep;20(3):577–596. [PubMed] [Google Scholar]
  6. Bland P. W., Warren L. G. Antigen presentation by epithelial cells of the rat small intestine. I. Kinetics, antigen specificity and blocking by anti-Ia antisera. Immunology. 1986 May;58(1):1–7. [PMC free article] [PubMed] [Google Scholar]
  7. Bland P. W., Whiting C. V. Antigen processing by isolated rat intestinal villus enterocytes. Immunology. 1989 Dec;68(4):497–502. [PMC free article] [PubMed] [Google Scholar]
  8. Bland P. W., Whiting C. V. Induction of MHC class II gene products in rat intestinal epithelium during graft-versus-host disease and effects on the immune function of the epithelium. Immunology. 1992 Feb;75(2):366–371. [PMC free article] [PubMed] [Google Scholar]
  9. Blumberg R. S., Balk S. P. Intraepithelial lymphocytes and their recognition of non-classical MHC molecules. Int Rev Immunol. 1994;11(1):15–30. doi: 10.3109/08830189409061714. [DOI] [PubMed] [Google Scholar]
  10. Blumberg R. S., Terhorst C., Bleicher P., McDermott F. V., Allan C. H., Landau S. B., Trier J. S., Balk S. P. Expression of a nonpolymorphic MHC class I-like molecule, CD1D, by human intestinal epithelial cells. J Immunol. 1991 Oct 15;147(8):2518–2524. [PubMed] [Google Scholar]
  11. Buus S., Werdelin O. A group-specific inhibitor of lysosomal cysteine proteinases selectively inhibits both proteolytic degradation and presentation of the antigen dinitrophenyl-poly-L-lysine by guinea pig accessory cells to T cells. J Immunol. 1986 Jan;136(2):452–458. [PubMed] [Google Scholar]
  12. Ceman S., Sant A. J. The function of invariant chain in class II-restricted antigen presentation. Semin Immunol. 1995 Dec;7(6):373–387. doi: 10.1006/smim.1995.0042. [DOI] [PubMed] [Google Scholar]
  13. Chang C. H., Flavell R. A. Class II transactivator regulates the expression of multiple genes involved in antigen presentation. J Exp Med. 1995 Feb 1;181(2):765–767. doi: 10.1084/jem.181.2.765. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Chen Y., Inobe J., Weiner H. L. Induction of oral tolerance to myelin basic protein in CD8-depleted mice: both CD4+ and CD8+ cells mediate active suppression. J Immunol. 1995 Jul 15;155(2):910–916. [PubMed] [Google Scholar]
  15. Ciclitira P. J., Nelufer J. M., Ellis H. J., Evans D. J. The effect of gluten on HLA-DR in the small intestinal epithelium of patients with coeliac disease. Clin Exp Immunol. 1986 Jan;63(1):101–104. [PMC free article] [PubMed] [Google Scholar]
  16. Cohen-Haguenauer O., Robbins E., Massart C., Busson M., Deschamps I., Hors J., Lalouel J. M., Dausset J., Cohen D. A systematic study of HLA class II-beta DNA restriction fragments in insulin-dependent diabetes mellitus. Proc Natl Acad Sci U S A. 1985 May;82(10):3335–3339. doi: 10.1073/pnas.82.10.3335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Cresswell P. Assembly, transport, and function of MHC class II molecules. Annu Rev Immunol. 1994;12:259–293. doi: 10.1146/annurev.iy.12.040194.001355. [DOI] [PubMed] [Google Scholar]
  18. Denzin L. K., Robbins N. F., Carboy-Newcomb C., Cresswell P. Assembly and intracellular transport of HLA-DM and correction of the class II antigen-processing defect in T2 cells. Immunity. 1994 Oct;1(7):595–606. doi: 10.1016/1074-7613(94)90049-3. [DOI] [PubMed] [Google Scholar]
  19. Diment S., Leech M. S., Stahl P. D. Cathepsin D is membrane-associated in macrophage endosomes. J Biol Chem. 1988 May 15;263(14):6901–6907. [PubMed] [Google Scholar]
  20. Furuhashi M., Nakahara A., Fukutomi H., Kominami E., Grube D., Uchiyama Y. Immunocytochemical localization of cathepsins B, H, and L in the rat gastro-duodenal mucosa. Histochemistry. 1991;95(3):231–239. doi: 10.1007/BF00266772. [DOI] [PubMed] [Google Scholar]
  21. Garside P., Steel M., Liew F. Y., Mowat A. M. CD4+ but not CD8+ T cells are required for the induction of oral tolerance. Int Immunol. 1995 Mar;7(3):501–504. doi: 10.1093/intimm/7.3.501. [DOI] [PubMed] [Google Scholar]
  22. Germain R. N. MHC-dependent antigen processing and peptide presentation: providing ligands for T lymphocyte activation. Cell. 1994 Jan 28;76(2):287–299. doi: 10.1016/0092-8674(94)90336-0. [DOI] [PubMed] [Google Scholar]
  23. Hashimoto Y., Komuro T. Close relationships between the cells of the immune system and the epithelial cells in the rat small intestine. Cell Tissue Res. 1988 Oct;254(1):41–47. doi: 10.1007/BF00220015. [DOI] [PubMed] [Google Scholar]
  24. Hoyne G. F., Callow M. G., Kuo M. C., Thomas W. R. Presentation of peptides and proteins by intestinal epithelial cells. Immunology. 1993 Oct;80(2):204–208. [PMC free article] [PubMed] [Google Scholar]
  25. Inoko H., Ando A., Tsuji K., Matsuki K., Juji T., Honda Y. HLA-DQ beta chain DNA restriction fragments can differentiate between healthy and narcoleptic individuals with HLA-DR2. Immunogenetics. 1986;23(2):126–128. doi: 10.1007/BF00377973. [DOI] [PubMed] [Google Scholar]
  26. Kaiserlian D., Vidal K., Revillard J. P. Murine enterocytes can present soluble antigen to specific class II-restricted CD4+ T cells. Eur J Immunol. 1989 Aug;19(8):1513–1516. doi: 10.1002/eji.1830190827. [DOI] [PubMed] [Google Scholar]
  27. Katz J. F., Stebbins C., Appella E., Sant A. J. Invariant chain and DM edit self-peptide presentation by major histocompatibility complex (MHC) class II molecules. J Exp Med. 1996 Nov 1;184(5):1747–1753. doi: 10.1084/jem.184.5.1747. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Kovats S., Drover S., Marshall W. H., Freed D., Whiteley P. E., Nepom G. T., Blum J. S. Coordinate defects in human histocompatibility leukocyte antigen class II expression and antigen presentation in bare lymphocyte syndrome. J Exp Med. 1994 Jun 1;179(6):2017–2022. doi: 10.1084/jem.179.6.2017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Kovats S., Nepom G. T., Coleman M., Nepom B., Kwok W. W., Blum J. S. Deficient antigen-presenting cell function in multiple genetic complementation groups of type II bare lymphocyte syndrome. J Clin Invest. 1995 Jul;96(1):217–223. doi: 10.1172/JCI118023. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Kwok W. W., Schwarz D., Nepom B. S., Hock R. A., Thurtle P. S., Nepom G. T. HLA-DQ molecules form alpha-beta heterodimers of mixed allotype. J Immunol. 1988 Nov 1;141(9):3123–3127. [PubMed] [Google Scholar]
  31. Lencer W. I., Strohmeier G., Moe S., Carlson S. L., Constable C. T., Madara J. L. Signal transduction by cholera toxin: processing in vesicular compartments does not require acidification. Am J Physiol. 1995 Oct;269(4 Pt 1):G548–G557. doi: 10.1152/ajpgi.1995.269.4.G548. [DOI] [PubMed] [Google Scholar]
  32. MacDonald M. J., Gottschall J., Hunter J. B., Winter K. L. HLA-DR4 in insulin-dependent diabetic parents and their diabetic offspring: a clue to dominant inheritance. Proc Natl Acad Sci U S A. 1986 Sep;83(18):7049–7053. doi: 10.1073/pnas.83.18.7049. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Madara J. L., Stafford J. Interferon-gamma directly affects barrier function of cultured intestinal epithelial monolayers. J Clin Invest. 1989 Feb;83(2):724–727. doi: 10.1172/JCI113938. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Mayer L., Eisenhardt D., Salomon P., Bauer W., Plous R., Piccinini L. Expression of class II molecules on intestinal epithelial cells in humans. Differences between normal and inflammatory bowel disease. Gastroenterology. 1991 Jan;100(1):3–12. doi: 10.1016/0016-5085(91)90575-6. [DOI] [PubMed] [Google Scholar]
  35. Mayer L., Shlien R. Evidence for function of Ia molecules on gut epithelial cells in man. J Exp Med. 1987 Nov 1;166(5):1471–1483. doi: 10.1084/jem.166.5.1471. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Mellins E., Smith L., Arp B., Cotner T., Celis E., Pious D. Defective processing and presentation of exogenous antigens in mutants with normal HLA class II genes. Nature. 1990 Jan 4;343(6253):71–74. doi: 10.1038/343071a0. [DOI] [PubMed] [Google Scholar]
  37. Mizuochi T., Yee S. T., Kasai M., Kakiuchi T., Muno D., Kominami E. Both cathepsin B and cathepsin D are necessary for processing of ovalbumin as well as for degradation of class II MHC invariant chain. Immunol Lett. 1994 Dec;43(3):189–193. doi: 10.1016/0165-2478(94)90221-6. [DOI] [PubMed] [Google Scholar]
  38. Momburg F., Koretz K., Von Herbay A., Möller P. Nonimmune human cells can express MHC class II antigens in the absence of invariant chain--an immunohistological study on normal and chronically inflamed small intestine. Clin Exp Immunol. 1988 Jun;72(3):367–372. [PMC free article] [PubMed] [Google Scholar]
  39. Panja A., Barone A., Mayer L. Stimulation of lamina propria lymphocytes by intestinal epithelial cells: evidence for recognition of nonclassical restriction elements. J Exp Med. 1994 Mar 1;179(3):943–950. doi: 10.1084/jem.179.3.943. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Pinet V., Malnati M. S., Long E. O. Two processing pathways for the MHC class II-restricted presentation of exogenous influenza virus antigen. J Immunol. 1994 May 15;152(10):4852–4860. [PubMed] [Google Scholar]
  41. Pinet V., Vergelli M., Martin R., Bakke O., Long E. O. Antigen presentation mediated by recycling of surface HLA-DR molecules. Nature. 1995 Jun 15;375(6532):603–606. doi: 10.1038/375603a0. [DOI] [PubMed] [Google Scholar]
  42. Powrie F. T cells in inflammatory bowel disease: protective and pathogenic roles. Immunity. 1995 Aug;3(2):171–174. doi: 10.1016/1074-7613(95)90086-1. [DOI] [PubMed] [Google Scholar]
  43. Roche P. A. HLA-DM: an in vivo facilitator of MHC class II peptide loading. Immunity. 1995 Sep;3(3):259–262. doi: 10.1016/1074-7613(95)90111-6. [DOI] [PubMed] [Google Scholar]
  44. Rodriguez G. M., Diment S. Role of cathepsin D in antigen presentation of ovalbumin. J Immunol. 1992 Nov 1;149(9):2894–2898. [PubMed] [Google Scholar]
  45. Sanderson I. R., Ouellette A. J., Carter E. A., Harmatz P. R. Ontogeny of class II MHC mRNA in the mouse small intestinal epithelium. Mol Immunol. 1992 Oct;29(10):1257–1263. doi: 10.1016/0161-5890(92)90062-3. [DOI] [PubMed] [Google Scholar]
  46. Sidhu N. K., Wright G. M., Markham R. J., Ireland W. P., Singh A. Quantitative regional variation in the expression of major histocompatibility class II antigens in enterocytes of the mouse small intestine. Tissue Cell. 1992;24(2):221–228. doi: 10.1016/0040-8166(92)90095-o. [DOI] [PubMed] [Google Scholar]
  47. Tuosto L., Karr R. W., Fu X. T., Olson R. R., Cundari E., Piccolella E., Lechler R., Lombardi G. Different regions of the N-terminal domains of HLA-DR1 influence recognition of individual peptide-DR1 complexes. Hum Immunol. 1994 Aug;40(4):312–322. doi: 10.1016/0198-8859(94)90031-0. [DOI] [PubMed] [Google Scholar]
  48. Vidal K., Samarut C., Magaud J. P., Revillard J. P., Kaiserlian D. Unexpected lack of reactivity of allogeneic anti-Ia monoclonal antibodies with MHC class II molecules expressed by mouse intestinal epithelial cells. J Immunol. 1993 Nov 1;151(9):4642–4650. [PubMed] [Google Scholar]
  49. Vidard L., Rock K. L., Benacerraf B. The generation of immunogenic peptides can be selectively increased or decreased by proteolytic enzyme inhibitors. J Immunol. 1991 Sep 15;147(6):1786–1791. [PubMed] [Google Scholar]
  50. Weiner G. J., Kaminski M. S. Anti-idiotypic antibodies recognizing stable epitopes limit the emergence of idiotype variants in a murine B cell lymphoma. J Immunol. 1990 Mar 15;144(6):2436–2445. [PubMed] [Google Scholar]
  51. Wolf P. R., Ploegh H. L. How MHC class II molecules acquire peptide cargo: biosynthesis and trafficking through the endocytic pathway. Annu Rev Cell Dev Biol. 1995;11:267–306. doi: 10.1146/annurev.cb.11.110195.001411. [DOI] [PubMed] [Google Scholar]
  52. Woods A., Chen H. Y., Trumbauer M. E., Sirotina A., Cummings R., Zaller D. M. Human major histocompatibility complex class II-restricted T cell responses in transgenic mice. J Exp Med. 1994 Jul 1;180(1):173–181. doi: 10.1084/jem.180.1.173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Yoshimori T., Yamamoto A., Moriyama Y., Futai M., Tashiro Y. Bafilomycin A1, a specific inhibitor of vacuolar-type H(+)-ATPase, inhibits acidification and protein degradation in lysosomes of cultured cells. J Biol Chem. 1991 Sep 15;266(26):17707–17712. [PubMed] [Google Scholar]
  54. Zhang Z. Y., Michael J. G. Orally inducible immune unresponsiveness is abrogated by IFN-gamma treatment. J Immunol. 1990 Jun 1;144(11):4163–4165. [PubMed] [Google Scholar]
  55. Zhong G., Romagnoli P., Germain R. N. Related leucine-based cytoplasmic targeting signals in invariant chain and major histocompatibility complex class II molecules control endocytic presentation of distinct determinants in a single protein. J Exp Med. 1997 Feb 3;185(3):429–438. doi: 10.1084/jem.185.3.429. [DOI] [PMC free article] [PubMed] [Google Scholar]

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