Advertisement

Annals of Hematology

, Volume 68, Issue 5, pp 237–239 | Cite as

Immune electron-microscopy study of the trimeric glycoprotein (CD 103) of hairy cell leukemia using the Ber-ACT8 monoclonal antibody

  • D. Frangoulidis
  • A. Schulz
  • H. Pralle
Original Article
  • 29 Downloads

Summary

We describe a method which makes it possible to study the expression of the trimeric glycoprotein (TGP, CD 103) on the surface of hairy cells (HCs) by electron microscopy (e.m.). The monoclonal antibody (mAb) Ber-ACT8 was used to identify the TGP. By testing mucosa of the small intestine, we found that a prefixation incubation with Ber-ACT8 was necessary. After isolation with a density gradient, Ber-ACT8 incubation, and fixation, the peroxidase-anti-peroxidase (PAP) method was used to make the TGP visible. The HCs showed a clearly discernible stain on their surface, a fairly constant distribution of the TGP, and a very good ultrastructural preservation. This method makes it possible to demonstrate the TGP or other unfixable antigens by e.m.

Key words

Immune electron microscopy Hairy cell leukemia Trimeric glycoprotein Ber-ACT8 monoclonal antibody 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Brunning RD, Parkin J (1975) Ribosome-lamella complexes in neoplastic hematopoietic cells. Am J Pathol 79:565–578Google Scholar
  2. 2.
    Cerf-Bensussan N, Jarry A, Brousse N et al. (1987) A monoclonal antibody (HML-1) defining a novel membrane molecule present on human intestinal lymphocytes. Eur J Immunol 17:1279–1285Google Scholar
  3. 3.
    Chandra S, Stefani SS (1978) Ultrastructure of granulo-filamentous bodies in pathologic spleen and lymph node. J Ultrastruct Res 64:148Google Scholar
  4. 4.
    Cordell JL, Falini B, Erber WN, Ghosh AK, Abdulaziz Z et al. (1984) Immunoenzymatic labeling of monoclonal antibodies using immune complexes of alkaline phosphatase and monoclonal anti-alkaline phosphatase (APAAP-Complexes). J Histochem Cytochem 32:219–229Google Scholar
  5. 5.
    Flenghi L, Spinozzi F, Stein H, Kruschwitz M, Pileri S, Falini R (1990) LF-61: a new monoclonal antibody directed against a trimeric molecule (150 kDa, 125 kDa, 105 kDa) associated with hairy cell leukaemia. Br J Haematol 76:451–459Google Scholar
  6. 6.
    Kruschwitz M, Fritsche G, Schwarting R, Micklem K, et al. (1991) Ber-ACT8: monoclonal antibody to the mucosa lymphocyte antigen. J Clin Pathol 44:635–645Google Scholar
  7. 7.
    Li CY, Ziesmer SC, Lazcano-Villareal O (1987) Use of azide and hydrogen peroxide as an inhibitor for endogenous peroxidase in the immunoperoxidase method. J Histochem Cytochem 35:1457–1460Google Scholar
  8. 8.
    Micklem KJ, Dong Y, Willis A, Pulford KA, Visser L et al. (1991) HML-1 antigen on mucosa-associated T cells, activated cells and hairy leukemic cells is a new integrin containing the β7 subunit. Am J Pathol 139:1297–1301Google Scholar
  9. 9.
    Möller P, Mielke B, Moldenhauer G (1990) Monoclonal antibody HML-1, a marker for intraepithelial T cells and lymphomas derived thereof, also recognizes hairy cell leukemia and some B-cell lymphomas. Am J Pathol 136:509–512Google Scholar
  10. 10.
    Moldenhauer G, Mielke B, Dörken B, Schwartz-Albiez R, Möller P (1990) Identity of HML-1 antigen on intestinal intraepithelial T cells and of B-ly7 antigen on hairy cell leukaemia. Scand J Immunol 32:77–82Google Scholar
  11. 11.
    Newman GR, Jasani B, Williams ED (1982) The preservation of ultrastructure and antigenicity. J Microscopy 127:RP5-RP6Google Scholar
  12. 12.
    Roberts AI, O'Connell SM, Ebert ECD (1993) Intestinal intraepithelial lymphocytes bind to colon cancer cells by HML-1 and CDlla. Cancer Res 53:1608–1611Google Scholar
  13. 13.
    Roberts K, Kilshaw PJ (1993) The mucosa T-cell integrin alpha (M 290) beta 7 recognizes a ligand on mucosal epithelial cell lines. Eur J Immunol 23:1630–1635Google Scholar
  14. 14.
    Schwarting R, Dienemann D, Kruschwitz M, Fritsche G, Stein H (1990) Specificities of monoclonal antibody B-ly7 and HML-1 are identical. Blood 75:320–321Google Scholar
  15. 15.
    Sternberger LA (1979) The unlabeled antibody peroxidaseanti-peroxidase (PAP) method. In: Sternberger LA (ed) Immunocytochemistry. Wiley and Sons, New York, pp 105–169Google Scholar
  16. 16.
    Visser L, Shaw A, Slupsky J, Vos H, Poppema S (1989) Monoclonal antibody reactive with hair cell leukemia. Blood 74:1Google Scholar
  17. 17.
    Willingham MC, Yamada SS (1979) Development of a new primary fixative for electron-microscopic immunocytochemical localization of intracellular antigens in cultured cells. J Histochem Cytochem 27:947–960Google Scholar

Copyright information

© Springer-Verlag 1994

Authors and Affiliations

  • D. Frangoulidis
    • 1
  • A. Schulz
    • 2
  • H. Pralle
    • 1
  1. 1.Department of Hematology and Oncology, Center for Internal MedicineJustus-Liebig UniversityGiessenGermany
  2. 2.Institute of PathologyJustus-Liebig UniversityGiessenGermany

Personalised recommendations