Multidimensional flow-cytometric analysis of dendritic cells in peripheral blood of normal donors and cancer patients.
We studied the potential of multidimensional flow cytometry to evaluate the frequency and maturation/activation status of dendritic cells in minimally manipulated peripheral blood mononuclear cell preparations (i.e., only separated on Ficoll-Hypaque) of normal donors and cancer patients. A rare subset of HLA-DR+ leukocytes (less than 1% mononuclear cells) was detected in blood of normal donors that displayed all the features of dendritic cells: these cells had high forward-light-scatter characteristics and coexpressed CD4, CD86 and CD54 surface antigens, but lacked the lineage-associated surface markers of T cells, B cells, monocytes, granulocytes or NK i.e. they were CD3-, CD19-, CD20-, CD14-, CD11b-, CD16-, CD56-). These physical and phenotypic properties were virtually identical to those of immunomagnetically sorted leukocytes characterized as dendritic-cells on the basis of morphology, phenotype and high stimulatory activity in allogeneic mixed-lymphocyte cultures. Using this flow-cytometric approach we observed that the frequency of dendritic cell-like cells in peripheral blood mononuclear cell specimens of cancer patients receiving chemotherapy alone or those recovering from stem cell transplantation was significantly lower than that of normal individuals (mean +/- SE: 0.36 +/- 0.05%, 0.14 +/- 0.06%, and 0.75 +/- 0.04% respectively). Multidimensional flow-cytometric analysis of dendritic cells might represent an important new tool for assessing immunocompetence, and for monitoring the effects of therapeutic regimens on the immune system.
Savary
C A
CA
Department of Surgical Oncology, University of Texas M.D. Anderson Cancer Center, Houston 77030, USA.
Grazziutti
M L
ML
Melichar
B
B
Przepiorka
D
D
Freedman
R S
RS
Cowart
R E
RE
Cohen
D M
DM
Anaissie
E J
EJ
Woodside
D G
DG
McIntyre
B W
BW
Pierson
D L
DL
Pellis
N R
NR
Rex
J H
JH
D003713:Dendritic Cells / Q000166:cytology* / Q000502:physiology; D005240:Feasibility Studies; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D018189:Immunomagnetic Separation; D007963:Leukocytes, Mononuclear / Q000166:cytology* / Q000502:physiology; D008027:Light; D010641:Phenotype; D012016:Reference Values; D012542:Scattering, Radiation