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Blood, Vol. 94 No. 7 (October 1), 1999:
pp. 2343-2356
The Monoclonal Antibody 97A6 Defines a Novel Surface Antigen Expressed
on Human Basophils and Their Multipotent and Unipotent Progenitors
Hans-Jörg Bühring,
Paul J. Simmons,
Melanie Pudney,
Robert Müller,
David Jarrossay,
Andreas van Agthoven,
Martin Willheim,
Wolfram Brugger,
Peter Valent, and
Lothar Kanz
From the Department of Medicine, Division of Hematology and Oncology,
University of Tübingen, Tübingen, Germany; Hanson Centre
for Cancer Research, Matthew Roberts Laboratory, Institute of Medical
and Veterinary Sciences, Adelaide, South Australia, Australia;
Immunotech, S.A., a Beckman-Coulter company, Marseille, France;
Department of Internal Medicine I, Division of Hematology and
Hemostaseology, University of Vienna, Vienna, Austria; and Institute of
General and Experimental Pathology, University of Vienna, Vienna,
Austria.
Basophils (Ba) and mast cells (MC) are important effector cells of
inflammatory reactions. Both cell types derive from CD34+
hematopoietic progenitors. However, little is known about the cell
subsets that become committed to and give rise to Ba and/or MC. We have
generated a monoclonal antibody (MoAb), 97A6, that specifically detects
human Ba, MC (lung, skin), and their CD34+ progenitors.
Other mature hematopoietic cells (neutrophils, eosinophils, monocytes,
lymphocytes, platelets) did not react with MoAb 97A6, and sorting of
97A6+ peripheral blood (PB) and bone marrow (BM) cells
resulted in an almost pure population (>98%) of Ba. Approximately
1% of CD34+ BM and PB cells was found to be
97A6+. Culture of sorted
CD34+97A6+ BM cells in semisolid medium
containing phytohemagglutinin-stimulated leukocyte
supernatant for 16 days (multilineage assay) resulted in the formation
of pure Ba colonies (10 of 40), Ba-eosinophil colonies (7 of 40),
Ba-macrophage colonies (3 of 40), and multilineage Ba-eosinophil-macrophage and/or neutrophil colonies (12 of 40). In
contrast, no Ba could be cultured from
CD34+97A6 cells. Liquid culture of
CD34+ PB cells in the presence of 100 ng/mL interleukin
(IL)-3 (Ba progenitor assay) resulted in an increase of
97A6+ cells, starting from 1% of day-0 cells to almost
70% (basophils) after day 7. Culture of sorted BM
CD34+97A6+ cells in the presence of 100 ng/mL stem cell factor (SCF) for 35 days (mast cell progenitor assay)
resulted in the growth of MC (>30% on day 35). Anti-IgE-induced IgE
receptor cross-linking on Ba for 15 minutes resulted in a 4-fold to
5-fold upregulation of 97A6 antigen expression. These data show that
the 97A6-reactive antigen plays a role in basophil activation and is
expressed on multipotent CD34+ progenitors, MC
progenitors, Ba progenitors, as well as on mature Ba and tissue MC. The
lineage-specificity of MoAb 97A6 suggests that this novel marker may be
a useful tool to isolate and analyze Ba/MC and their progenitors.
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