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 L. Haviland, Ph.D.	     Internet:"haviland at"   +
+  Washington Univ. School of Med.   A.K.A : The Compiler                 +
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