anti-mouse CD ab fail to react with wood mice blood cells

Tony Schountz tschount at mesastate.edu
Fri Mar 1 12:47:43 EST 2002


This is probably to be expected. Although Apodemus and Mus (and Rattus)
are all murids, there is likely significant differences in these genes.
We tested 17 monoclonal antibodies specific for Mus proteins for
cross-reactivity to deer mouse (Peromyscus maniculatus) leukocytes, but
only 3 gave acceptable results. These were to CD45R-B220 (RA3-6B2),
CD62L (MEL-14), and CD43 (S7), all from Pharmingen. We had inconclusive
results with an anti-CD80 antibody (16-10A1). Peromyscus (sigmodontine)
is 25-50 million years divergent (myd) from Mus and some genes exhibit a
lot of divergence at the amino acid level. CD19, for example, is so
divergent between species that it is difficult to design primers from
conserved regions in orthologs to amplify and clone partial sequences
from other species. Here are the antibodies we tried, but did not work:

Specificity (clone)
CD3e (145-2C11)
CD4 (RM4-4)
CD8a (53-6.7)
CD11a (M17/4)
CD11b (M1/70)
CD11c (HL3)
CD19 (1D3)
CD21/35 (7G6)
CD24 (M1/69)
CD86 (GL1)
Ig kappa (R8-140)
IgD 'a' allele (AMS9.1)
TCR beta (H57-597)

I don't know the divergence of Apodemus from Mus or Rattus, but Mus and
Rattus are 5-10 myd. Most of the antibodies we tested were produced in
the rat. If yours are too, and if Rattus and Apodemus are closer than
either is to Mus, it is probable that some of your antibodies won't work
because rats and wood mice will share determinants, therefore rats won't
make antibodies that bind to the epitopes on wood mouse CD molecules.

On Thursday, February 28, 2002, at 08:33  AM, Van Bockstaele, Dirk wrote:

>
> Hi Flowers,
>
> I recently got involved in a collaboration with scientists studying the
> effect of environmental pollutants by investigating rodents (wood mice)
> they
> catch in the field.
> In preliminary experiments I was surprised to see that some monoclonals
> against mice CD's (3/19/11b/13) that show normal reactivity towards lab
> mice
> (Mus musculus) leukocytes fail to do so towards blood leukocytes of the
> wood
> mice (apodemus sylvaticus).
>
> My question: is this to be expected or not?  Is a lab mouse so different
> from a wood mouse?  Or should I complain with the mab manufacturer
> and/or
> start looking for another manufacturer?  I'm a complete novice
> regarding lab
> mice work let alone wood mice!  Any hints, suggestions are very welcome!
> thanks in advance.
> Dirk
>
> Prof. Dirk Van Bockstaele
> Laboratory of Hematology
> Head of Flow Cytometry / Molecular Diagnostics
> Antwerp University Hospital
> Wilrijkstraat 10
> B-2650 Edegem
> Belgium
> phone 32 3 821 3900 fax 32 3 825 1148
>
>
--
Tony Schountz, Ph.D.
Dept. of Biology
Mesa State College
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