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J Vet Sci. 2008 Mar;9(1):51-66 |
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Use of flow cytometry to develop and characterize a set of monoclonal antibodies specific for rabbit leukocyte differentiation molecules
William C. Davis*, Mary Jo Hamilton |
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Department of Veterinary Microbiology and Pathology, College of Veterinary Medicine, Washington State University, Pullman, WA 99164-7040, USA
* davisw@vetmed.wsu.edu |
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Flow cytometry was used to identify and characterize
monoclonal antibodies (mAbs) that react with rabbit
leukocyte differentiation molecules (LDM). Screening sets
of mAbs, developed against LDM in other species, for
reactivity with rabbit LDM yielded 11 mAbs that
recognize conserved epitopes on rabbit LDM orthologues
and multiple mAbs that recognize epitopes expressed on
the major histocompatibility class I or class II molecules.
Screening of mAbs submitted to the Animal Homologues
Section of the Eighth Human Leukocyte Differentiation
Workshop yielded 7 additional mAbs. Screening of mAbs
generated from mice immunized with leukocytes from
rabbit thymus or spleen or concanavalin A activated
peripheral blood and/or spleen lymphocytes has yielded 42
mAbs that recognize species restricted epitopes expressed
on one or more lineages of leukocytes. Screening of the
anti-rabbit mAbs against leukocytes from other species
yielded one additional mAb. The studies show that screening
of existing sets of mAbs for reactivity with rabbit LDM
will not be productive and that a direct approach will be
needed to develop mAbs for research in rabbits. The flow
cytometric approach we developed to screen for mAbs of
interest offers a way for individual laboratories to identify
and characterize mAbs to LDM in rabbits and other
species. A web-based program we developed provides a
source of information that will facilitate analysis. It
contains a searchable data base on known CD molecules
and a data base on mAbs, known to react with LDM in
one or more species of artiodactyla, equidae, carnivora,
and or lagomorpha.
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