CD107a-tetramer double staining

Sandra Tuyaerts Sandra.Tuyaerts at vub.ac.be
Tue Nov 8 07:13:42 EST 2005


Hello everybody,
I want to ask a question about the following:
We encounter a problem when we want to test antigen-specific T cells
both for tetramer positivity and CD107a positivity (measure of
degranulation).
We restimulate the T cells with T2 cells pulsed with either an
irrelevant or the appropriate peptide in the presence of golgi inhibitor
and CD107a antibody. After 6 hours or overnight co-culture we perform
tetramer and CD8 staining on these cells. The T cells restimulated with
irrelevant peptide-pulsed T2 cells show nice tetramer positivity and low
background staining for CD107a. The T cells restimulated with relevant
peptide-pulsed T2 cells show a nice CD107a positivity, but unfortunately
these cells don't stain positive with the tetramers anymore. 
What could have happened during this co-culture that the cells become
tetramer negative? Can anybody give us a hint? 
Thanks in advance,
Sandra	
Sandra Tuyaerts
Laboratorium voor Moleculaire en Cellulaire Therapie
Vakgroep Fysiologie - Immunologie
Laarbeeklaan 103/E
1090 Jette (Belgium)
Tel: 02/477.45.11.
Fax: 02/477.45.68.
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