Two distinct cell populations are obtained from human blood monocytes cultured with M-CSF, GM-CSF and IL-4

1999 ◽  
Vol 35 ◽  
pp. S39-S40 ◽  
Author(s):  
C.L Baron ◽  
S.M Scholl ◽  
H Bausinger ◽  
D Hanau ◽  
P Pouillart ◽  
...  
Keyword(s):  
Human Blood ◽  
Cell Populations ◽  
Blood Monocytes ◽  
Gm Csf ◽  
Distinct Cell

scholarly journals In-VitroDifferentiation of Mature Dendritic Cells From Human Blood Monocytes

1998 ◽  
Vol 6 (1-2) ◽  
pp. 25-39 ◽  
Author(s):  
Robert Gieseler ◽  
Dirk Heise ◽  
Afsaneh Soruri ◽  
Peter Schwartz ◽  
J. Hinrich Peters
Keyword(s):  
Dendritic Cells ◽  
Human Blood ◽  
T Cell Response ◽  
Blood Monocytes ◽  
Gm Csf ◽  
Hla Dr ◽  
Hla Dq ◽  
Specific Stimulation ◽  
Antigen Presenting

Representing the most potent antigen-presenting cells, dendritic cells (DC) can now be generated from human blood monocytes. We recently presented a novel protocol employing GM-CSF, IL-4, and IFN-γto differentiate monocyte-derived DCin vitro. Here, such cells are characterized in detail. Cells in culture exhibited both dendritic and veiled morphologies, the former being adherent and the latter suspended. Phenotypically, they were CD1a-/dim, CD11a+, CD11b++, CD11c+, CD14dim/-, CD16a-/dim, CD18+, CD32dim/-, CD33+, CD40+, CD45R0+, CD50+, CD54+, CD64-/dim, CD68+, CD71+, CD80dim, CD86+/++, MHC class I++/+++HLA-DR++/+++HLA-DP+, and HLA-DQ+. The DC stimulated a strong allogeneic T-cell response, and further evidence for their autologous antigen-specific stimulation is discussed. Although resembling a mature CD 11c+CD45R0+blood DC subset identified earlier, their differentiation in the presence of the Thl and Th2 cytokines IFN-γand IL-4 indicates that these DC may conform to mature mucosal DC.

scholarly journals Azithromycin drives in vitro GM-CSF/IL-4-induced differentiation of human blood monocytes toward dendritic-like cells with regulatory properties

2011 ◽  
Vol 91 (2) ◽  
pp. 229-243 ◽  
Author(s):  
Darija Stupin Polančec ◽  
Vesna Munić Kos ◽  
Mihailo Banjanac ◽  
Mila Vrančić ◽  
Snježana Čužić ◽  
...  
Keyword(s):  
Human Blood ◽  
Blood Monocytes ◽  
Induced Differentiation ◽  
Gm Csf ◽  
Regulatory Properties

A Role for Notch Signaling in the Commitment of Human Blood Monocytes to Langerhans Cells.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 3452-3452
Author(s):  
Tetsunori Shibasaki ◽  
Naoyuki Katayama ◽  
Kohshi Ohishi ◽  
Masahiro Masuya ◽  
Hiroshi Shiku
Keyword(s):  
Notch Signaling ◽  
Human Blood ◽  
Langerhans Cells ◽  
Cultured Cells ◽  
Early Stage ◽  
Expression Level ◽  
Blood Monocytes ◽  
Notch Ligand ◽  
Gm Csf ◽  
Tgf Β1

Abstract We previously demonstrated that Notch ligand Delta-1 in concert with GM-CSF and TGF-β1 promotes the differentiation of human blood monocytes into Langerhans cells that are characterized by the expression of CD1a, Langerhans-associated granules Langerin, cutaneous lymphocyte-associated antigen (CLA), CC chemokine receptor 6 (CCR6), and E-cadherin. These data extended the functional scope of Notch ligand Delta-1 in human adult hematopoiesis. HES-1 is known to be the target gene by Notch signaling. We examined the effect of Delta-1 on the expression of HES-1 mRNA in CD14+ blood monocytes in the presence of GM-CSF and TGF-β1, using real-time RT-PCR. When CD14+ blood monocytes were cultured with Delta-1, GM-CSF, and TGF-β1, the expression level of HES-1 mRNA increased approximately 10-fold at 24 hours of incubation, compared with the expression level in freshly isolated CD14+ monocytes. However, the expression level of HES-1 mRNA declined at 48 hours of incubation. This finding suggests that Delta-1 may operate at the early stage of the differentiation pathway from CD14+ monocytes to Langerhans cells. To explore this issue more precisely, we cultured CD14+ monocytes in the presence of Delta-1, GM-CSF, and TGF-β1 for 2 days, and subsequently replated the cells into the cultures without Delta-1. At day 7 of culture, cultured cells were harvested and characterized by phenotypic analysis. This initial 2-day exposure of CD14+ monocytes to Delta-1 gave rise to Langerhans cells, similar to the observation obtained with the supplementation of Delta-1 throughout 7-day culture. In turn, when Delta-1 was added at day 2 of culture, Langerhans cells were not induced, but instead the resulting cells exhibited the features of macrophages. Our results indicate that in response to Delta-1, human blood monocytes appear to initiate the differentiation program toward Langerhans cells, while they are incapable of differentiating into Langerhans cells after their commitment to macrophages.

An Indispensable Role of GM-CSF in the Differentiation of Human Blood Monocytes toward Langerhans Cells.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 3868-3868
Author(s):  
Tetsunori Shibasaki ◽  
Naoyuki Katayama ◽  
Kohshi Ohishi ◽  
Fumihiko Monma ◽  
Zhaocai Yu ◽  
...  
Keyword(s):  
Human Blood ◽  
Langerhans Cells ◽  
Cultured Cells ◽  
Expression Profiles ◽  
Blood Monocytes ◽  
Similar Action ◽  
Gm Csf ◽  
Tgf Β1 ◽  
Cells Cultured ◽  
E Cadherin

Abstract We recently demonstrated that GM-CSF, TGFβ1, and Notch ligand Delta-1, all of which are synthesized in the milieu of the skin, instruct human blood monocytes to differentiate into Langerhans cells (LCs) that are characterized by the expression of CD1a, Langerin, CLA, CCR6, and E-cadherin and the presence of Birbeck granules. Previous studies have shown similar biologic activities of GM-CSF and IL-3 on human blood monocytes. In the present study, we investigated whether GM-CSF and IL-3 have a similar action on blood monocytes with respect to their differentiation into LCs. In contrast to CD14+ monocytes cultured for 7 days in the presence of GM-CSF, TGF-β1, and Delta-1, the cells obtained from culture with IL-3, TGF-β1, and Delta-1 were negative for CLA, CCR6, Langerin, and E-cadherin. These cells expressed HLA-ABC, HLA-DR, CD80, CD86, CD40, CD54, and CD11c but not DC-SIGN. The expression level of CD1a declined, while CD14 was upregulated. The difference between cells cultured with GM-CSF, TGF-β1, and Delta-1 and those with IL-3, TGF-β1, and Delta-1 was corroborated by microarray analysis of the gene expression profiles. Thus, GM-CSF and IL-3 exert distinct effects on human blood monocytes in the presence of TGF-β1 and Delta-1. When GM-CSF was added to cultures containing IL-3, TGF-β1, and Delta-1, the phenotype of cultured cells closely resembled that observed with GM-CSF, TGF-β1, and Delta-1. As GM-CSF and IL-3 share common receptor β subunits, the signals mediated through GM-CSF receptor α subunits appear to be required for the development of LCs from monocytes. We also compared the action of GM-CSF with that of IL-3 in terms of the differentiation of monocytes into macrophages and dendritic cells (DCs). When CD14+ monocytes were cultured in the presence of GM-CSF or IL-3 for 7 days, the resulting macrophages obtained from both cultures were phenotypically indistinguishable. Next, we examined whether IL-3 as well as GM-CSF induces CD14+ monocytes to differentiate into DCs in the presence of IL-4. The phenotypic profiles in cells cultured with IL-3 plus IL-4 were parallel to those seen with DCs cultured in the presence of GM-CSF plus IL-4. These data suggest that although IL-3 can substitute for GM-CSF in the differentiation pathway of CD14+ monocytes toward macrophages or DCs, GM-CSF is indispensable for their LC development.

Moribund Blood Monocytes: Apoptosis Limits the Yield of Monocyte Derived Dentritic Cells from Apheresis Samples.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 3894-3894
Author(s):  
Lars Macke ◽  
Robert Geffers ◽  
Manfred Rohde ◽  
Henk S. Garritsen ◽  
Bernhard Woermann ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
Large Fraction ◽  
Flow Cytometric Analysis ◽  
Cell Populations ◽  
Dna Arrays ◽  
Blood Monocytes ◽  
Apoptosis Pathway ◽  
Gm Csf ◽  
Negative Population ◽  
Survival Signals

Abstract Cells of the monocyte/macrophage/dendritic cell (DC) lineage play a central role in inflammation and form the bridge between innate and acquired immunity. In recent years monocytes also found special attention as source of precursor cells for the generation of monocyte derived dendritic cells for immunotherapy. However, monocytes circulate in the blood stream only temporarily with a half life of about 24 h and die spontaneously from apoptosis in the absence of specific survival signals. When we investigated the survival of blood monocytes isolated by CD14 positive selection from apheresis samples and cultivated in the presence of GM-CSF/IL4, we observed a loss of viability of about 66% of the cells within 48h. This would be consistent with the view that about 80% of the monocytes are bound to die at the time of isolation. Detailed flow cytometric analysis revealed that loss of vital cells is accompanied by the appearance of a CD14-negative population, which stained positive for annexin V. To analyse the fate of these cell populations CD14-positive and CD14-negative cells were separated by cell sorting at day 1 and day 2. Flow cytometry and electron microscopy identified predominantly dying cells in the CD14-negative population, whereas a large fraction of the CD14-positive population survived. The CD14-positive cells can be differentiated with an efficiency of 64%–91% to mature DC after cultivation under DC generating conditions; these include the presence of GM-CSF and IL4 and addition of a maturation mix (TNFa, IL1b, IL6 and PGE2) on day 5. DC generated from sorted CD14-positive cell populations were qualitatively and quantitatively comparable to DC from unsorted monocytes. To study the genes involved we analyzed the transcriptome on each cultivation day using Affymetrix HG-U133A DNA-Arrays. During the first 24h, the expressions of about 4700 genes were regulated. About 3000 genes were up regulated and about 1700 genes were down regulated. Especially, we focused on the expression of genes involved in the apoptosis pathway during the early phase of cultivation. Among apoptosis related genes caspase 3, a central player in natural apoptosis of monocytes, was up regulated on day 1. In summary these data suggest that apoptosis limits the yield of monocyte derived DC and that cultivation in the presence of caspase inhibitors or survival signals could improve the efficiency of generation of monocyte derived dendritic cells.

Lymphokine-activated human blood monocytes destroy tumor cells but not normal cells under cocultivation conditions.

1984 ◽  
Vol 2 (8) ◽  
pp. 937-943 ◽  
Author(s):  
I J Fidler ◽  
E S Kleinerman
Keyword(s):  
Human Blood ◽  
Target Cell ◽  
Cell Populations ◽  
Blood Monocytes ◽  
Neoplastic Cells ◽  
Multilamellar Liposomes ◽  
Release Assay ◽  
Normal Human ◽  
Activated Monocytes

The possibility that tumoricidal human blood monocytes would recognize and destroy tumorigenic targets but leave bystander nontumorigenic cells unharmed was investigated. Highly purified preparations of peripheral blood monocytes isolated from normal human donors were activated in vitro by incubation with human lymphokines encapsulated in multilamellar liposomes. The cytotoxic properties of these monocytes against several tumorigenic and nontumorigenic allogeneic target cell populations were assessed by an in vitro radioisotope-release assay. Various combinations of three tumorigenic and three nontumorigenic target-cell populations, labeled with either [3H]thymidine or [14C]thymidine, were mixed and plated onto monolayers of blood monocytes. In all combinations used, activated monocytes specifically lysed only allogeneic neoplastic cells. At least in vitro, activated human blood monocytes can recognize and selectively destroy neoplastic cells.

IL-4 and IL-10 synergistically inhibit survival of human blood monocytes supported by GM-CSF

2005 ◽  
Author(s):  
Hiroyuki Miyashita ◽  
Naoyuki Katayama ◽  
Atsushi Fujieda ◽  
Tetsunori Shibasaki ◽  
Kentaro Yamamura ◽  
...  
Keyword(s):  
Human Blood ◽  
Blood Monocytes ◽  
Gm Csf

scholarly journals Independent regulation of M-CSF and G-CSF gene expression in human monocytes

Blood ◽  
1988 ◽  
Vol 71 (6) ◽  
pp. 1529-1532 ◽  
Author(s):  
E Vellenga ◽  
A Rambaldi ◽  
TJ Ernst ◽  
D Ostapovicz ◽  
JD Griffin
Keyword(s):  
Gene Expression ◽  
Progenitor Cells ◽  
Human Blood ◽  
Human Monocytes ◽  
Blood Monocytes ◽  
Colony Stimulating Factors ◽  
Interleukin 3 ◽  
Gm Csf ◽  
Proliferation And Differentiation

The macrophage and granulocyte colony-stimulating factors, M-CSF and G- CSF, act in vitro to induce proliferation and differentiation of monocyte and granulocyte progenitor cells, respectively. We show here that both of these CSFs can be produced by stimulated human blood monocytes, but the M-CSF and G-CSF genes are independently regulated. Recombinant human interleukin-3 (IL-3) and GM-CSF primarily induce expression of the M-CSF gene and secretion of M-CSF, whereas bacterial lipopolysaccharide primarily induces expression of the G-CSF gene and secretion of G-CSF. These results suggest that under different conditions of in vitro stimulation the monocyte secretes factors that could lead selectively to either granulocyte or monocyte production.

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