Elaine,
Altman JD. Moss PAH. Goulder PJR. Barouch DH. McHeyzer-Williams MG. Bell JI. McMichael AJ. Davis MM.
Phenotypic analysis of antigen-specific T lymphocytes
Science. 274(5284):94-6, 1996 Oct 4.
Good luck,
Tony Bakke
Antony Bakke, PhD
Oregon Health Sci Univ
Dept of Pathology
bakkea@ohsu.edu
>>> Elaine Kunze <mek4@psu.edu> 04/26 7:54 AM >>>
One of my faculty came back from a meeting saying "All the immunologist do
a tetramer assay for antigen presenting cells."
I feel silly but Tetramer assays are a very new method of measuring cytotoxic T cells
or helper T cells, depending on the MHC class used. A specific peptide
is used so the T cells are antigen specific. These studies have shown
that original estimates of antigen specific T cells by limiting dilution
are 100-1000 fold low. However, the method is not trivial. You have
to synthesize soluble MHC molecules and bind these to specific antigens
with the right motif for that MHC. Four MHC-antigen complexes are held
together by biotin on the MHC that is bound to fluorescent avidin.
Once you have the reagent, the assay is easy. I would certainly not
call it routine and it is NOT used to measure antigen presenting cells.
John Altman in Mark Davis' lab first described the technique, so you
could search on Davis and Altman. One of the first papers is: I believe
that John Altman has been very helpful in working with other labs to
produce MHC tetramers and I have heard a rumor that Coulter might try to
offer them commercially, but that is only a distant rumor. So don't be
surprised that not everyone knows about this assay and tell your faculty
member that he might look into making the tetramers (only joking).
I never heard of this and didn't find anything after a
quicky look at Current protocols and a few other generic sources. Can
anyone provide a reference and or explanation?
****************************************************************************
Elaine Kunze
Flow Cytometry.....Image Analysis...
The Biotechnology Institute for Research and Education
Life Sciences Consortium
8B Althouse Laboratory (814-863-2762)
Penn State University
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