HELP! Why are my T cells normal by FACS?????

Tom Esch tre at wista.wistar.upenn.edu
Fri Aug 26 11:04:45 EST 1994


In article <33id2j$drg at mercury.hgmp.mrc.ac.uk>, p-donoho at nimr.mrc.ac.uk
(Paul M. Donohoe) wrote:
> 
> I am examining the changes in immune system parameters over a time
> course in rats with Adjuvant Arthritis, and in r65kD-protected rats,
> one part of which is looking at lymphocyte surface markers by
> immunostaining and FACS....
[snip--basically, there's no difference between normal and arthritic rats]

Paul--
     My experience with flow cytometry supports what you have found 
concerning temporary storage of cells before staining, and keeping 
them on ice for a few hours should make no difference to your
results. I think it more likely that either 1) the surface antigens
you're staining for don't show any difference between normal and 
diseased animals or 2) the cells you're looking at aren't the
population that show any changes that might occur. The first 
possibility seems less likely to me based on the little I know
about arthritis--in your list of markers IL2-R and MHC class II
(at least) stand out as ones that should show some difference in
activated T cells, and thus (presumably) in cells from the 
diseased rats. Have you tried getting lymphocytes out of the 
inflamed joints and doing a similar (parallel?) analysis? I'm
somewhat surprised that cells from the reactive nodes you describe
show no change from normal, but I'd be *shocked* if cells from the
joints looked the same--they would seem to be your ultimate
positive control. Also, what do similar studies in humans show? Are
there visible changes in peripheral blood lymphocytes that reflect
arthritis activity? My admittedly poor memory says that there are,
but I can't remember exactly what. IL2-R and class II would be good
guesses, though. I hope there's some help for you somewhere in all
this.            --Tom


"I do not know, Sir, if the fellow is an infidel; but if he be an 
infidel, he is an infidel as a dog is an infidel; that is, he has 
never thought upon the subject."  --Johnson



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