Partial degradation of T-cell signal transduction molecules by contaminating granulocytes during protein extraction of splenic T cells from tumor-bearing mice.
Cancer Res
; 55(17): 3840-6, 1995 Sep 01.
Article
in En
| MEDLINE
| ID: mdl-7641202
ABSTRACT
Flavone-8-acetic acid plus recombinant human interleukin 2 is a successful antitumor therapy in mice bearing the Renca murine renal cell carcinoma. This report demonstrates that T cells, particularly CD8+ T cells, are critical for the generation of this response. Initial experiments examining T-cell signal transduction proteins demonstrated that T cells from Renca-bearing mice had undetectable levels of p56lck and zeta-chain of the T-cell receptor and that flavone-8-acetic acid and recombinant human interleukin 2 therapy could be used as a model for reversal of these alterations. However, further experimentation showed that the majority of the reduction in zeta-chain and part of the reduction in p56lck was due to degradation of these molecules during protein extraction caused by mature granulocytes contaminating the enriched T-cell population. This was not the case for nuclear c-Rel or NF kappa B p65, which remained at undetectable/reduced levels in the absence of granulocytes, confirming our previous data that transcription factor alterations exist in tumor-bearing mice. Thus, most of the reduction in zeta-chain in T cells from Renca-bearing mice is due to granulocyte contamination and emphasizes the need to use pure T-cell populations and/or sufficient amounts and types of protease inhibitors when quantitating proteins in T cells from tumor-bearing mice.
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Collection:
01-internacional
Database:
MEDLINE
Main subject:
Flavonoids
/
Protein-Tyrosine Kinases
/
Carcinoma, Renal Cell
/
Receptors, Antigen, T-Cell
/
CD4-Positive T-Lymphocytes
/
Signal Transduction
/
Interleukin-2
/
CD8-Positive T-Lymphocytes
/
Granulocytes
/
Immunotherapy
Type of study:
Prognostic_studies
Limits:
Animals
Language:
En
Journal:
Cancer Res
Year:
1995
Document type:
Article
Affiliation country:
United States