Non-activating CD's on T cells

Dr Stephen Young (steve@rheuma.bham.ac.uk)
Fri, 15 Jan 93 9:52:23 GMT

Using the our Coulter Elite FACS we can investigate Ca fluxes in lymphocytes
using Indo-1 and this seems to work quite well now we have found a source
of reliable indo (Molecular Probes). We have found some interesting results
using whole PBMC, PBL and purified T cells. The latter were obtained by negative
selection, as we are very concerned about pre-activation of the cells prior
to use, and possibly inducing anergy. Now we have this method established
we would like to be able to look at some subsets within the T cells and also
check that the responding cells are T cells. For this we would like to surface
label the cells, but its increasing clear that Ab crosslinking of many CD's
can induce a signal.

Has anyone any experience of using surface labelling that does not activate?
This could either be through non-signalling CD's ( for instance CD7 has been
suggested as a possibility) or using Fab fragments. There also arises the
problem of which fluorophores to use and still allow indo-1 ratioing to be
done.

Any suggestions would be appreciated.

Steve Young
Dept of Rheumatology
University of Birmingham

Email to s.p.young@bham.ac.uk Voice: 021-414-6480

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