Skip to main content
PMC home page
The American Journal of Pathology logoLink to The American Journal of Pathology
. 1987 Oct;129(1):54–63.

Immunophenotyping of non-Hodgkin's lymphomas using a panel of antibodies on paraffin-embedded tissues.

F R Davey 1, K C Gatter 1, E Ralfkiaer 1, K A Pulford 1, G W Krissansen 1, D Y Mason 1
PMCID: PMC1899695  PMID: 3310651

Abstract

The use of monoclonal and polyclonal antibodies for the immunophenotyping of non-Hodgkin's lymphomas in paraffin-embedded tissue has been limited by the fact that most antigens on lymphoid cells are denatured by histologic fixation, dehydration, and embedment. In this article the authors have analyzed a small panel of antibodies which represent exceptions to this rule, in that they identify denaturation-resistant determinants on leukocyte antigens in paraffin-embedded tissue. Monoclonal antibodies L26 [corrected] and 4KB5 label preferentially B cells, monoclonal antibody UCHL1 stains predominantly T cells, and monoclonal antibody MAC 387 reacts with granulocytes and some macrophages. A polyclonal antiserum raised against purified CD3 (T3) antigen, a T-cell-specific molecule, was also employed. This antibody panel was used to immunophenotype routinely processed tissue biopsy specimens from 61 non-Hodgkin's lymphomas (all of which had been previously phenotyped in cryostat sections). The lineage of the neoplastic cells was correctly identified in 32 of 34 (94%) cases of B-cell lymphoma, in 19 of 19 (100%) cases of T-cell neoplasm, and in 2 of 4 (50%) cases of histiocytic malignancy. It is concluded that this combination of antibodies is helpful in immunophenotyping non-Hodgkin's lymphomas when only paraffin-embedded tissue sections are available, although additional reagents of higher specificity are required to improve the identification of lymphomas.

Full text

PDF
62

Images in this article

57Figure 1
on p.57
58Figure 2
on p.58
59Figure 3
on p.59
60Figure 4
on p.60
61Figure 5
on p.61

Selected References

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

  1. Borowitz M. J., Bousvaros A., Brynes R. K., Cousar J. B., Crissman J. D., Whitcomb C. C., Kerns B. J., Byrne G. E., Jr Monoclonal antibody phenotyping of B-cell non-Hodgkin's lymphomas. The Southeastern Cancer Study Group experience. Am J Pathol. 1985 Dec;121(3):514–521. [PMC free article] [PubMed] [Google Scholar]
  2. Borowitz M. J., Croker B. P., Metzgar R. S. Comparison of histologic and immunologic heterogeneity of non-Hodgkin's lymphomas. Am J Pathol. 1981 Nov;105(2):97–106. [PMC free article] [PubMed] [Google Scholar]
  3. Cordell J. L., Falini B., Erber W. N., Ghosh A. K., Abdulaziz Z., MacDonald S., Pulford K. A., Stein H., Mason D. Y. Immunoenzymatic labeling of monoclonal antibodies using immune complexes of alkaline phosphatase and monoclonal anti-alkaline phosphatase (APAAP complexes). J Histochem Cytochem. 1984 Feb;32(2):219–229. doi: 10.1177/32.2.6198355. [DOI] [PubMed] [Google Scholar]
  4. Dalchau R., Fabre J. W. Identification with a monoclonal antibody of a predominantly B lymphocyte-specific determinant of the human leukocyte common antigen. Evidence for structural and possible functional diversity of the human leukocyte common molecule. J Exp Med. 1981 Apr 1;153(4):753–765. doi: 10.1084/jem.153.4.753. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Epenetos A. A., Bobrow L. G., Adams T. E., Collins C. M., Isaacson P. G., Bodmer W. F. A monoclonal antibody that detects HLA-D region antigen in routinely fixed, wax embedded sections of normal and neoplastic lymphoid tissues. J Clin Pathol. 1985 Jan;38(1):12–17. doi: 10.1136/jcp.38.1.12. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Epstein A. L., Marder R. J., Winter J. N., Fox R. I. Two new monoclonal antibodies (LN-1, LN-2) reactive in B5 formalin-fixed, paraffin-embedded tissues with follicular center and mantle zone human B lymphocytes and derived tumors. J Immunol. 1984 Aug;133(2):1028–1036. [PubMed] [Google Scholar]
  7. Franklin W. A., Mason D. Y., Pulford K., Falini B., Bliss E., Gatter K. C., Stein H., Clarke L. C., McGee J. O. Immunohistological analysis of human mononuclear phagocytes and dendritic cells by using monoclonal antibodies. Lab Invest. 1986 Mar;54(3):322–335. [PubMed] [Google Scholar]
  8. Hsu S. M., Jaffe E. S. Leu M1 and peanut agglutinin stain the neoplastic cells of Hodgkin's disease. Am J Clin Pathol. 1984 Jul;82(1):29–32. doi: 10.1093/ajcp/82.1.29. [DOI] [PubMed] [Google Scholar]
  9. Ishii Y., Takami T., Yuasa H., Takei T., Kikuchi K. Two distinct antigen systems in human B lymphocytes: identification of cell surface and intracellular antigens using monoclonal antibodies. Clin Exp Immunol. 1984 Oct;58(1):183–192. [PMC free article] [PubMed] [Google Scholar]
  10. Knowles D. M., 2nd, Halper J. P. Human T-cell malignancies: Correlative clinical, histopathologic, immunologic, and cytochemical analysis of 23 cases. Am J Pathol. 1982 Feb;106(2):187–203. [PMC free article] [PubMed] [Google Scholar]
  11. Lukes R. J., Parker J. W., Taylor C. R., Tindle B. H., Cramer A. D., Lincoln T. L. Immunologic approach to non-Hodgkin lymphomas and related leukemias. Analysis of the results of multiparameter studies of 425 cases. Semin Hematol. 1978 Oct;15(4):322–351. [PubMed] [Google Scholar]
  12. Marder R. J., Variakojis D., Silver J., Epstein A. L. Immunohistochemical analysis of human lymphomas with monoclonal antibodies to B cell and Ia antigens reactive in paraffin sections. Lab Invest. 1985 May;52(5):497–504. [PubMed] [Google Scholar]
  13. Mepham B. L., Frater W., Mitchell B. S. The use of proteolytic enzymes to improve immunoglobulin staining by the PAP technique. Histochem J. 1979 May;11(3):345–357. doi: 10.1007/BF01005033. [DOI] [PubMed] [Google Scholar]
  14. Norton A. J., Ramsay A. D., Smith S. H., Beverley P. C., Isaacson P. G. Monoclonal antibody (UCHL1) that recognises normal and neoplastic T cells in routinely fixed tissues. J Clin Pathol. 1986 Apr;39(4):399–405. doi: 10.1136/jcp.39.4.399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Poppema S., Hollema H., Visser L., Vos H. Monoclonal antibodies (MT1, MT2, MB1, MB2, MB3) reactive with leukocyte subsets in paraffin-embedded tissue sections. Am J Pathol. 1987 Jun;127(3):418–429. [PMC free article] [PubMed] [Google Scholar]
  16. Shimizu M., Miura M., Kitahara M., Katayama I. Non-Hodgkin's lymphomas. analysis of 109 Japanese cases with the use of LSGJ classification. Am J Pathol. 1982 Jan;106(1):30–39. [PMC free article] [PubMed] [Google Scholar]
  17. Smith S. H., Brown M. H., Rowe D., Callard R. E., Beverley P. C. Functional subsets of human helper-inducer cells defined by a new monoclonal antibody, UCHL1. Immunology. 1986 May;58(1):63–70. [PMC free article] [PubMed] [Google Scholar]
  18. Stass S. A., Schumacher H. R., Keneklis T. P., Bollum F. J. Terminal deoxynucleotidyl transferase immunofluorescence of bone marrow smears: experience in 156 cases. Am J Clin Pathol. 1979 Dec;72(6):898–903. doi: 10.1093/ajcp/72.6.898. [DOI] [PubMed] [Google Scholar]
  19. Stein H., Gerdes J., Schwab U., Lemke H., Mason D. Y., Ziegler A., Schienle W., Diehl V. Identification of Hodgkin and Sternberg-reed cells as a unique cell type derived from a newly-detected small-cell population. Int J Cancer. 1982 Oct 15;30(4):445–459. doi: 10.1002/ijc.2910300411. [DOI] [PubMed] [Google Scholar]
  20. Stein H., Lennert K., Feller A. C., Mason D. Y. Immunohistological analysis of human lymphoma: correlation of histological and immunological categories. Adv Cancer Res. 1984;42:67–147. doi: 10.1016/s0065-230x(08)60456-x. [DOI] [PubMed] [Google Scholar]
  21. Tsan M. F., McIntyre P. A. Stimulation by propylthiouracil of the hexose monophosphate shunt in human polymorphonuclear leucocytes during phagocytosis. Br J Haematol. 1975 Oct;31(2):193–208. doi: 10.1111/j.1365-2141.1975.tb00850.x. [DOI] [PubMed] [Google Scholar]
  22. Warnke R. A., Gatter K. C., Falini B., Hildreth P., Woolston R. E., Pulford K., Cordell J. L., Cohen B., De Wolf-Peeters C., Mason D. Y. Diagnosis of human lymphoma with monoclonal antileukocyte antibodies. N Engl J Med. 1983 Nov 24;309(21):1275–1281. doi: 10.1056/NEJM198311243092102. [DOI] [PubMed] [Google Scholar]
  23. West K. P., Warford A., Fray L., Allen M., Campbell A. C., Lauder I. The demonstration of B-cell, T-cell and myeloid antigens in paraffin sections. J Pathol. 1986 Oct;150(2):89–101. doi: 10.1002/path.1711500203. [DOI] [PubMed] [Google Scholar]

Articles from The American Journal of Pathology are provided here courtesy of American Society for Investigative Pathology

RESOURCES