Anti-TCR cross-reactivity?

David L. Haviland, Ph.D. HAVILAND at KIDS.WUSTL.EDU
Wed Oct 19 15:29:54 EST 1994


Greetings netters:

I'm afilliated with the Children's Hospital Clinical Immunology lab and I
have an interesting problem on which I'd like to solicit the groups
thoughts.  We have been expanding our diagnostic flow cytometry panel for T
and B cells, adhesion molecules, etc...  For whatever CD marker we examine,
we have been very successful in using whole blood for our staining.  In the
analysis, the RBCs/platelet are simply gated out on the basis of size.  As
I said, this works for any "CD-whatever you want" antigen EXCEPT when we
look specifically at the T-cell receptor (alpha/beta-specifically).  Direct
staining on whole blood doesn't work with this antibody - T cells are
negative for TCR!  Yet, when the lymphocytes are isolated via Ficoll, they
of course stain for the TCR.  I had the (senior) lab tech to do multiple
gating on *anything* that scatters light (FALS and 90') and it turns out
that the RBCs were absorbing the TCR MAb.  Why? 

The only thing I can come up with is some sort of non-specific carbohydrate 
association between the antibody and the variety of carbohydrate moities on 
RBCs.  I'm not sure if this thought "holds any water"?

Any thoughts? ...and many thanks in advance.
David
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