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Isolation of human blood dendritic cells using the CMRF-44 monoclonal
antibody: implications for studies on antigen-presenting cell function and
immunotherapy
DB Fearnley, AD McLellan, SI Mannering, BD Hock and DN Hart
Haematology/Immunology Research Group, Christchurch Hospital, New Zealand.
Dendritic cells (DC) are potent antigen-presenting cells (APC) with the
capacity to stimulate a primary T lymphocyte immune response and are
therefore of interest for potential immunotherapeutic applications. Freshly
isolated DC or DC precursors may be preferable for studies of antigen
uptake and the potential control of APC costimulator activity. In this
report, we report that the monoclonal antibody CMRF-44 can be used to
detect early DC differentiation. The majority of DC circulating in blood do
not express any known DC lineage specific markers, but can be identified by
CMRF-44 labeling after a brief period of in vitro culture. The sequential
acquisition of DC activation antigens allows the identification of two
stages of DC maturation/activation. Cytokines, especially
granulocyte-macrophage colony-stimulating factor (GM-CSF) and tumor
necrosis factor (TNF)alpha, enhance both phases of this process, whereas
CD40-ligand trimer preferentially enhances the final DC maturation to a
fully mature, activated phenotype. DC positively selected using CMRF-44
possess potent allostimulatory activity and are efficient at the uptake,
processing, and presentation of soluble antigens for both primary and
secondary immune responses. CMRF-44+ DC are also more potent than other APC
types at restimulation of a chronic myeloid leukemia peptide specific
T-cell clone. The use of a purified population of freshly isolated DC may
be advantageous in attempts to initiate, maintain, and direct immune
responses for immunotherapeutic applications.
Volume 89,
Issue 10,
pp. 3708-3716,
05/15/1997
Copyright © 1997 by The American Society of Hematology
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