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. 1989 Nov;42(11):1194–1200. doi: 10.1136/jcp.42.11.1194

Detection of T cells in paraffin wax embedded tissue using antibodies against a peptide sequence from the CD3 antigen.

D Y Mason 1, J Cordell 1, M Brown 1, G Pallesen 1, E Ralfkiaer 1, J Rothbard 1, M Crumpton 1, K C Gatter 1
PMCID: PMC501981  PMID: 2531171

Abstract

Rabbit polyclonal antibodies were raised against a proline rich, peptide sequence, comprising 13 amino acids, in the cytoplasmic domain of the CD3 epsilon chain. Immunoprecipitation experiments showed that this antibody preparation recognised the CD3 antigen on human T lymphoblasts. The antibody stained normal T cells strongly in tissue sections which had been fixed in formalin or Bouin's solution and embedded in paraffin wax. Its reactivity with T cell lymphoma, when evaluated on a series of 96 previously phenotyped cases, closely agreed with the results obtained on cryostat sections. These results indicate that the specific detection of T cells in routinely processed tissue biopsy specimens is now technically feasible on a wide scale in diagnostic laboratories using CD3 peptide antibodies, and they also suggest that in future the use of anti-peptide antibodies may detect other lineage specific antigenic markers in paraffin wax sections.

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Selected References

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

  1. Brown M. H., Sewell W. A., Mason D. Y., Rothbard J. B., Crumpton M. J. Species conservation of the T cell lymphocyte CD2 cell surface antigen. Eur J Immunol. 1988 Aug;18(8):1223–1227. doi: 10.1002/eji.1830180812. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Garson J. A., Beverley P. C., Coakham H. B., Harper E. I. Monoclonal antibodies against human T lymphocytes label Purkinje neurones of many species. Nature. 1982 Jul 22;298(5872):375–377. doi: 10.1038/298375a0. [DOI] [PubMed] [Google Scholar]
  4. Gold D. P., Puck J. M., Pettey C. L., Cho M., Coligan J., Woody J. N., Terhorst C. Isolation of cDNA clones encoding the 20K non-glycosylated polypeptide chain of the human T-cell receptor/T3 complex. Nature. 1986 May 22;321(6068):431–434. doi: 10.1038/321431a0. [DOI] [PubMed] [Google Scholar]
  5. Hall P. A., D'Ardenne A. J., Butler M. G., Habeshaw J. R., Stansfeld A. G. New marker of B lymphocytes, MB2: comparison with other lymphocyte subset markers active in conventionally processed tissue sections. J Clin Pathol. 1987 Feb;40(2):151–156. doi: 10.1136/jcp.40.2.151. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hastrup N., Ralfkiaer E., Pallesen G. Aberrant phenotypes in peripheral T cell lymphomas. J Clin Pathol. 1989 Apr;42(4):398–402. doi: 10.1136/jcp.42.4.398. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Mason D. Y., Krissansen G. W., Davey F. R., Crumpton M. J., Gatter K. C. Antisera against epitopes resistant to denaturation on T3 (CD3) antigen can detect reactive and neoplastic T cells in paraffin embedded tissue biopsy specimens. J Clin Pathol. 1988 Feb;41(2):121–127. doi: 10.1136/jcp.41.2.121. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Myskow M. W., Krajewski A. S., Salter D. M., Dobson C. M., Miller E. P. Paraffin section immunophenotyping of non-Hodgkin's lymphoma, using a panel of monoclonal antibodies. Am J Clin Pathol. 1988 Nov;90(5):564–574. doi: 10.1093/ajcp/90.5.564. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. 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]
  12. Pulido R., Cebrián M., Acevedo A., de Landázuri M. O., Sánchez-Madrid F. Comparative biochemical and tissue distribution study of four distinct CD45 antigen specificities. J Immunol. 1988 Jun 1;140(11):3851–3857. [PubMed] [Google Scholar]
  13. Rothbard J. B., Fernandez R., Schoolnik G. K. Strain-specific and common epitopes of gonococcal pili. J Exp Med. 1984 Jul 1;160(1):208–221. doi: 10.1084/jem.160.1.208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Rothbard J. B., Lechler R. I., Howland K., Bal V., Eckels D. D., Sekaly R., Long E. O., Taylor W. R., Lamb J. R. Structural model of HLA-DR1 restricted T cell antigen recognition. Cell. 1988 Feb 26;52(4):515–523. doi: 10.1016/0092-8674(88)90464-3. [DOI] [PubMed] [Google Scholar]
  15. Stross W. P., Warnke R. A., Flavell D. J., Flavell S. U., Simmons D., Gatter K. C., Mason D. Y. Molecule detected in formalin fixed tissue by antibodies MT1, DF-T1, and L60 (Leu-22) corresponds to CD43 antigen. J Clin Pathol. 1989 Sep;42(9):953–961. doi: 10.1136/jcp.42.9.953. [DOI] [PMC free article] [PubMed] [Google Scholar]

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