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Granted, in a thymidine incorporation assay, not all of the cells that are
incorporating label are antigen specific either. I would like to get an idea
how others in the field are grappling with this issue.
Calman Prussin
Allergic Diseases Section
National Institute of Allergy and Infectious Diseases
National Institutes of Health
Bethesda, Maryland 20892
calman_prussin@nih.gov
------------------------ Forwarded Message -----------------------
Date: 6-30-95 2:49pm
From: {brettT@qimr.edu.au}:unix:niaid
To: calman prussin:10:niaid
Subj: lymph.function assay by CD69 and CD71.
Also-to: cytometry mailing list <cytometry@flowcyt.cyto.purdue.edu>
Also-to: brettt@qimr.edu.au
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We have just started using a CD69 and CD71 based lymphocyte function
assay (based on some work I've previously done and the BDIS paper in Cytometry
20:127-133 (1995)).
Generally I am seeking a group opinion on whether this method is valid
to replace the tritiated Thymidine method, and, what ways can it be improved
to make it more viable an alternative.
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.
Brett Teale
Queensland Institute of Medical Research/Dept.of Immunology,
Royal Brisbane Hospital
Queensland
Australia
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