Histochemistry

, Volume 101, Issue 5, pp 355–363

The ER-TR4 monoclonal antibody recognizes murine thymic epithelial cells (Type 1) and inhibits their capacity to interact with immature thymocytes: immuno-electron microscopic and functional studies

  • M.-P. Defresne
  • E. Van Vliet
  • R. Willemsen
  • H. Van Dongen
  • W. Van Ewijk
  • B. Nabarra
Original Paper

Abstract

The thymic stroma is heterogeneous with regard to cellular morphology and cellular function. In this study, we employed the monoclonal antibody ER-TR4 to characterize stromal cells at the ultrastructural level. To identify the labelled cell type, we used two techniques: immunogold labelling on ultrathin frozen sections and immunoperoxidase staining on thick “vibratome” sections. ER-TR4 reacted with thymic Type 1 epithelial cells (according to our classification). A dense labelling appears in the cytoplasm of cortical cells using the two techniques. Immunogold labelling identified small cytoplasmic vesicles whereas the cytoplasm and the cell membrane seem to be labelled with the immunoperoxidase technique. ER-TR4 also identified isolated thymic nurse cells (TNC), and was observed in vitro to inhibit the capacity of some type 1 epithelial cells to establish interactions with immature thymocytes. This finding supports the hypothesis that the factor is involved in the formation of lymphoepithelial interactions within thymic nurse cells, and thus in the relations that immature thymocytes establish with the thymic microenvironment.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Andrews P, Boyd RL (1985) The murine thymic nurse cell: an isolated thymic microenvironment. Eur J Immunol 15: 36–42Google Scholar
  2. Carding SR, Hayday AC, Bottomly K (1991) Cytokines in T-cell development. Immunol Today 12: 239–245Google Scholar
  3. De Waal Malefijt R, Leene W, Roholl P, Wormmeester J, Hoeben K (1986) T cell differentiation within thymic nurse cells. Lab Invest 55: 25–34Google Scholar
  4. Defresne MP, Goffinet G, Boniver J (1986) In situ characterization in freeze-fractured mouse thymuses of lymphoepithelial complexes ultrastructurally similar to isolated thymic nurse cells. Tissue Cell 18: 321–330Google Scholar
  5. Defresne MP, Humblet C, Rongy AM, Greimers R, Boniver J (1990) Effect of interferon-gamma and tumor necrosis factoralpha on lymphoepithelial interactions within thymic nurse cells. Eur J Immunol 20: 429–432Google Scholar
  6. Deman J, Martin MT, Delvenne P, Humblet C, Boniver J, Defresne MP (1992) Analysis by in situ hybridization of cells expressing mRNA for tumor necrosis factor in the developing thymus of mice. Dev Immunol 2: 103–109Google Scholar
  7. Duijvestijn A, Hoefsmit E (1981) Ultrastructure of the rat thymus. The microenvironment of T lymphocyte maturation. Cell Tissue Res 218: 279–282Google Scholar
  8. Farr A, Nelson A, Twex J, Hosier S (1991) Epithelial heterogeneity in the murine thymus: a cell surface glycoprotein expressed by subcapsular and medullary epithelium. J Histochem Cytochem 39: 645–653Google Scholar
  9. Fowlkes BJ, Pardoll DW (1989) Molecular and cellular events of T cell development. Adv Immunol 44: 207–264Google Scholar
  10. Geenen V, Defresne MP, Robert F, Legros JJ, Franchimont P, Boniver J (1988) The neurohormonal thymic microenvironment: immunocytochemical evidence that thymic nurse cells are neuroendocrine cells. Neuroendocrinology 47: 365–368Google Scholar
  11. Haynes B (1984) The human thymic microenvironment. Adv Immunol 36: 87–142Google Scholar
  12. Haynes B, Shimizy K, Eisenbarth G (1983) Identification of human and rodent thymic epithelium using tetanus toxin and monoclonal antibody A2B5. J Clin Invest 71: 9–17Google Scholar
  13. Hirokawa K, Utsuyama M, Morrizumi H, Handa S (1986) Analysis of the thymic microenvironment by monoclonal antibodies with special reference to thymic nurse cells. Thymus 8: 349–360Google Scholar
  14. Houben-Defresne MP, Varlet A, Goffinet G, Boniver J (1982) Thymic nurse cells are the first site of virus replication after inoculation of the radiation leukemia virus. Leuk Res 6: 231–241Google Scholar
  15. Kendall MD, Ritter MA, (eds) (1988) Thymus update, vol 1. The microenvironment of the human thymus. Harwood Academic Publishers, SwitzerlandGoogle Scholar
  16. Kyewski BA (1991) Thymic nurse cells: possible sites of T-cell selection. Immunol Today 7: 374–379Google Scholar
  17. Kyewski BA, Kaplan HS (1982) Lymphoepithelial interactions in the mouse thymus: phenotypic and kinetic studies on thymic nurse cells. J Immunol 128: 2287–2294Google Scholar
  18. Kyewski BA, Rouse RV, Kaplan HS (1982) Thymocyte rosettes: multicellular complexes of lymphocytes and bone marrow derived cells in the mouse thymus. Proc Natl Acad Sci USA 79: 5646–5650Google Scholar
  19. Marrack P, Brinster R, Palmiter R, Burkly L, Flavell R, Kappler J (1988) The effect of thymus environment on T cell development and tolerance. Cell 53: 627–634Google Scholar
  20. Nabarra B, Andrianarison I (1987) Ultrastructural studies of thymic reticulum. I. Epithelial component. Thymus 9: 95–121Google Scholar
  21. Nabarra B, Andrianarison I (1991) Ultrastructural studies of thymic reticulum. II. Non-epithelial component. Thymus 17: 39–61Google Scholar
  22. Penninger J, Hala K, Wick G (1990) Intrathymic nurse cell lymphocytes can induce a specific graft-versus-host-reaction. J Exp Med 172: 521–529Google Scholar
  23. Ramsdell F, Fowlkes BJ (1990) Clonal deletion versus clonal anergy — the role of the thymus in inducing self-tolerance. Science 248: 1342–1348Google Scholar
  24. Rouse R, Bolin L, Bender J, Kyewski BA (1988) Monoclonal antibodies reactive with subsets of mouse and human thymic epithelial cells. J Histochem Cytochem 36: 1511–1517Google Scholar
  25. Singer K (1990) Interactions between epithelial cells and T lymphocytes: role of adhesion molecules. J Leuk Biol 48: 367–374Google Scholar
  26. Singer K, Denning S, Whichard L, Haynes B (1990) Thymocyte LFA-1 and thymic epithelial cell ICAM/1 molecules mediate binding of activated human thymocytes to thymic epithelial cells. J Immunol 144: 2931–2939Google Scholar
  27. Stutman O (1978) Intrathymic and extrathymic cell maturation. Immunol Rev 422: 138–184Google Scholar
  28. Tokuyasu KT (1973) A technique for ultracryotomy of cell suspensions and tissues. J Cell Biol 57: 551–565Google Scholar
  29. Tokuyasu KT (1984) Immuno-cryoultramicrotomy. In: Polak JM, Varndell IM (eds) Immunolabelling for electron microscopy. Elsevier, Science Publishers, Amsterdam, pp 71–82Google Scholar
  30. Toussaint-Demylle D, Scheiff JM, Haumont S (1990) Thymic nurse cells: morphological study during their isolation from murine thymus. Cell Tissue Res 261: 115–123Google Scholar
  31. Van Ewijk W (1988) Cell surface topography of thymic microenvironments. Lab Invest 59: 579–590Google Scholar
  32. Van Ewijk W (1989) Immunohistology of lymphoid and non lymphoid cells in the thymus in relation to T lymphocyte differentiation. Am J Anat 170: 311–330Google Scholar
  33. Van Ewijk W (1991) T-cell differentiation is influenced by thymic microenvironments. Annu Rev Immunol 9: 591–615Google Scholar
  34. Van Ewijk W, Coffman R, Weissman IL (1980a) Immunoelectron microscopy of cell surface antigen: a quantitative analysis of antibody binding after different fixation protocols. Histochem J 12: 349–358Google Scholar
  35. Van Ewijk W, Rouse RV, Weissman IL (1980b) Distribution of H-2 microenvironments in the mouse thymus. J Histochem Cytochem 28: 1089–1099Google Scholar
  36. Van Vliet E, Melis M, Van Ewijk W (1984) Monoclonal antibodies to stromal cell types of the mouse thymus. Eur J Immunol 14: 524–529Google Scholar
  37. Van Vliet E, Melis M, Van Ewijk W (1984b) Immunohistology of thymic nurse cells. Cell Immunol 87: 101–109Google Scholar
  38. Wekerle H, Ketelsen U, Ernst M (1980) Thymic nurse cells. Lymphoepithelial cell complexes in murine thymuses: morphological and serological characterization. J Exp Med 151: 925–944Google Scholar

Copyright information

© Springer Verlag 1994

Authors and Affiliations

  • M.-P. Defresne
    • 1
  • E. Van Vliet
    • 2
  • R. Willemsen
    • 2
  • H. Van Dongen
    • 2
  • W. Van Ewijk
    • 2
  • B. Nabarra
    • 3
  1. 1.Laboratory of Pathological AnatomyUniversity of LiègeLiègeBelgium
  2. 2.Department of Cell Biology and GeneticsErasmus UniversityRotterdamThe Netherlands
  3. 3.U345 INSERMHôpital NeckerParisFrance

Personalised recommendations