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The method uses ficolled lymphocytes (1million) stimulated for 4 hours
with PWM, Con A, CD2, PHA, SEB, Candida A., PMA as a maximal control and an
unstimulated control. These are then FACSed 4 hours using CD3/CD69 for
activation and again at 16hours with CD3/CD71 for proliferation.
My Questions:
1. Is there a need for an additional proliferation marker (eg. CD25) or is
CD71 alone acceptable.
2. Would more co-stimulatory mitogens (eg. CD2 and 28) be more relevant
clinically to test activation/function than mitogens like PWM and ConA.
3. Would DNA profiles be of any real advantage in this case.
4 Are there any conditions/disorders where this assay may not perform.
and
5. Am I just chasing my tail, or is this truly an alternative to the tritiated
thymidine.
Brett Teale
Queensland Institute of Medical Research/Dept.of Immunology,
Royal Brisbane Hospital
Queensland
Australia
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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web