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Bruce H. Davis, M.D.
Wm. Beaumont Hospital
810-551-5137
FAX 810-551-8057
On Mon, 3 Jul 1995, Eric Miller wrote:
>
>
> On Fri, 30 Jun 1995 brettT@qimr.edu.au wrote:
>
> >
> >
> > 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.
> >
> >
> > 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.
>
> My only comment on this would be that BrDU incorporation would probably
> give the closest correlation with tritiated thymidine, and would also
> give you a DNA profile to compare results with if you wished.
> > Brett Teale
> > Queensland Institute of Medical Research/Dept.of Immunology,
> > Royal Brisbane Hospital
> > Queensland
> > Australia
> >
> > brettT@qimr.edu.au
>
> Eric Miller, ICRF,Edinburgh UK
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