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Characterization of Monoclonal Antibodies That Recognize Canine CD34
Peter A. McSweeney,
Katherine A. Rouleau,
Philip M. Wallace,
Benedetto Bruno,
Robert G. Andrews,
Ljiljana Krizanac-Bengez,
Brenda M. Sandmaier,
Rainer Storb,
Elizabeth Wayner, and
Richard A. Nash
From the Clinical Research Division, Fred Hutchinson Cancer Research
Center, Seattle, WA; the University of Washington, Seattle, WA; and
Bristol-Myers Squibb Pharmaceutical Research Institute, Seattle, WA.
Using a polyclonal antiserum against canine CD34, we previously
found that CD34 is expressed on canine bone marrow progenitor cells in
a manner analogous to that found in humans. To further characterize
CD34+ cells and to facilitate preclinical canine stem
cell transplant studies, monoclonal antibodies (MoAbs) were raised to
CD34. A panel of 10 MoAbs was generated that reacted with recombinant CD34 and with CD34+ cell lines and failed to react with
CD34 cell lines. Binding properties of five purified
MoAbs were determined by BIAcore analysis and flow cytometric staining,
and several MoAbs showed high affinity for CD34. Two antibodies, 1H6
and 2E9, were further characterized, and in flow cytometry studies
typically 1% to 3% of stained bone marrow cells were
CD34+. Purified CD34+ bone marrow cells
were 1.8- to 55-fold enriched for colony-forming unit-granulocyte-macrophage and for long-term culture initiating cells
as compared with bone marrow mononuclear cells, whereas CD34 cells were depleted of progenitors. Three
autologous transplants were performed with CD34+ cell
fractions enriched by immunomagnetic separation. After marrow ablative
total body irradiation (920 cGy), prompt hematopoietic recovery was
seen with transplanted cell doses of  1.1 × 107 /kg
that were 29% to 70% CD34+. Engraftment kinetics were
similar to those of dogs previously transplanted with approximately 10- to 100-fold more unmodified autologous marrow cells. This suggests that
CD34+ is a marker not only of canine bone marrow
progenitors but also for cells with radioprotective or marrow
repopulating function in vivo. MoAbs to CD34 will be
valuable for future studies of canine hematopoiesis and preclinical
studies concerning stem cell transplantation, gene therapy, and ex vivo
progenitor cell expansion.
Blood, Vol. 91 No. 6 (March 15), 1998:
pp. 1977-1986
© 1998 by The American Society of Hematology.
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