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Cytoplasmic expression of the CD3 antigen as a diagnostic marker for
immature T-cell malignancies
JJ van Dongen, GW Krissansen, IL Wolvers-Tettero, WM Comans-Bitter, HJ Adriaansen, H Hooijkaas, ER van Wering and C Terhorst
Department of Cell Biology, Immunology, and Genetics, Erasmus University,
Rotterdam, The Netherlands.
The expression of cytoplasmic CD3 (CyCD3) was analyzed in 45 leukemias,
five thymus cell samples, five peripheral blood (PB) samples, and ten cell
lines. All T cell acute lymphoblastic leukemias (T-ALL) that did not
express surface membrane CD3 (SmCD3) appeared to express CyCD3.
Furthermore, the majority of SmCD3+ T-ALL also expressed CyCD3. Analogous
results were obtained with thymus cell samples in that about 95% of the
thymocytes expressed CyCD3 whereas 60% to 75% of the thymocytes also
expressed SmCD3. In normal peripheral blood only prominent SmCD3 expression
was found. These data indicate that immature T cells express CyCD3 only,
that the combined expression of CyCD3 and SmCD3 is characteristic for
intermediate differentiation stages, and that mature T cells express
prominent SmCD3. All (precursor) B cell leukemias, acute myeloid leukemias,
and non-T cell lines tested did not express CyCD3. On the basis of these
data, we conclude that CyCD3 expression is restricted to the T cell lineage
and can be used as a diagnostic marker for immature SmCD3- T cell
malignancies. Therefore, we evaluated which fixative is optimal for CyCD3
staining, and we determined by immunofluorescence staining and Western
blotting which anti-CD3 monoclonal antibody (MoAb) can be used for the
detection of CyCD3. In our opinion, acid ethanol was the best fixative for
the cytocentrifuge preparations. Furthermore, we demonstrated that CyCD3
can be easily detected by use of MoAbs raised against denaturated CD3
chains such as those of the SP series (SP-6, SP-10, SP-64, and SP-78). In
addition we tested 22 anti-CD3 MoAbs of the Oxford CD3 panel that were
raised against native SmCD3, and it appeared that only four (UCHT1, VIT-3b,
G19-41 and SK7/Leu-4) of them were able to detect CyCD3. In Western blot
analysis all four MoAbs recognized the CD3- epsilon chain only.
Volume 71,
Issue 3,
pp. 603-612,
03/01/1988
Copyright © 1988 by The American Society of Hematology
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