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IMMUNOBIOLOGY
From the Walter and Eliza Hall Institute of Medical
Research, Melbourne, Australia; Institute for Medical Microbiology,
Immunology and Hygiene, Technical University of Munich, Germany; and
Pharmacia, St Louis, MO.
We studied the effects of administration of several cytokines,
including progenipoietin-1 (ProGP-1), Flt-3 ligand (FL), granulocyte colony-stimulating factor (G-CSF), and granulocyte-macrophage colony-stimulating factor in a pegylated form (pGM-CSF), on dendritic cell (DC) populations in mouse spleen. ProGP-1 produced the most striking increase in overall DC numbers, apparently more than its
constituent FL and G-CSF components. However, the expansion in DC
numbers was strongly subpopulation selective, with ProGP-1 and FL
producing selective expansion of CD8+ DCs, whereas pGM-CSF
produced selective expansion of CD8 Dendritic cells (DCs) are potent antigen-presenting
cells with a unique ability to activate naive T cells.1,2
They are distributed throughout lymphoid and nonlymphoid tissues but
are relatively rare, constituting less than 1% of spleen cells. DCs show heterogeneity in surface phenotype, origin, and some functional characteristics.3-8 We previously identified 3 distinct DC
populations, delineated by the expression of CD4 and CD8
(CD4+CD8 DCs are candidate targets for immune system modulation, particularly
for immunotherapy of tumors. One strategy used to target DCs
specifically is ex vivo manipulation of these cells, such as loading
DCs with tumor antigens before administration to the patient.12,13 Another strategy is in vivo administration
of cytokines that selectively increase DC numbers. Cytokines that have
been found to increase DC numbers dramatically in mice include Flt-3
ligand (FL), used alone14,15 or with CD40L,16
and granulocyte-macrophage colony-stimulating factor (GM-CSF) in a
polyethylene glycol-conjugated or pegylated form (pGM-CSF) that
extends its life span in the circulation.17,18 In humans,
DC numbers were increased by administering FL19 or
granulocyte colony-stimulating factor (G-CSF).20 In addition, progenipoietin 1 (ProGP-1), an agonist of the Flt-3 and G-CSF
receptors, markedly enhanced DC numbers in mice.21 However, the various DC populations are not absolutely dependent on
GM-CSF or FL for development. In the absence of GM-CSF receptor or
GM-CSF itself, little change occurs in the number or phenotype of
splenic DCs.18,22 Although DC numbers are reduced in FL knockout mice, DCs of both CD8 If expansion of DC populations by in vivo cytokine administration is to
be exploited for therapy, it is essential to understand the changes in
DC subpopulation balance and the characteristics of expanded DCs
relative to those of normal DCs. Accordingly, we compared the phenotype
and function of DCs expanded by ProGP-1, FL of mouse (muFL) and human
(huFL) origin, G-CSF, FL together with G-CSF, and pGM-CSF. We found
that certain cytokines induced marked differential expansion of
functionally distinct DC subsets and that some cytokines caused changes
in the cytokine-secretion capacity of these DCs.
Mice and in vivo administration of cytokines
Cytokines
DC stimulants Fixed and heat-killed Staphylococcus aureus (SAC; Pansorbin) was purchased from Calbiochem-Novabiochem (Alexandria, Australia). Oligonucleotides containing a fully phosphorothioated (ph) CpG motif were synthesized by GeneWorks (Adelaide, Australia) according to a published sequence (CpG166824).Isolation of DCs DCs were isolated as described previously.4 Briefly, spleen fragments were digested for 20 minutes at room temperature with collagenase and deoxyribonuclease and then treated for 5 minutes with EDTA to disrupt T-cell-DC complexes. Light-density cells were selected by centrifugation over Nycodenz medium (1.077 g/cm3, murine osmolarity). When DCs were purified from mice treated with FL or ProGP-1, the procedure was modified and 10 mL Nycodenz was used for each spleen equivalent to avoid overloading the separation medium. Cells not of DC lineage were depleted by first incubating the cells with titrated levels of anti-CD3 (KT3), anti-Thy-1 (T24/31.7), anti-B220 (RA3-6B2), anti-Gr-1 (RB6-8C5), and antierythrocyte (Ter-119) monoclonal antibodies (mAbs). The antibody-coated cells were then removed by incubation with sheep anti-rat IgG-coupled magnetic beads (Dynabeads M-450; Dynal, Oslo, Norway) or goat anti-rat IgG-coupled magnetic beads (Paesel and Lorei, Duisburg, Germany). The DCs prepared in this way were at least 85% pure at this stage. Immunofluorescent labeling and then sorting or gating during flow cytometry were used to complete purification of the DCs.Immunofluorescent labeling of DCs The mAbs, fluorescent conjugates, and multicolor labeling procedures were all described previously.4 To identify and sort all DCs, the pan-DC markers used were high levels of class II major histocompatibility complex (MHC), CD11c, or both, together with high forward light scatter. Anti-CD11c (N418) was used as a Cy5 or fluorescein isothiocyanate (FITC) conjugate. Anti-class II MHC (N22 or M5/114) was used as a FITC or Alexa 594 conjugate; conjugation levels were deliberately kept at less than maximal to ensure that the strong staining for class II MHC at saturation did not cause inaccurate color-compensation problems in other channels. The markers used to separate the splenic DC subpopulations were CD8 and CD4. Anti-CD8
(YTS169.4) was used as a phycoerythrin (PE) or Cy5 conjugate and
anti-CD4 (GK1.5) as a PE or Alexa 594 conjugate. The staining for
costimulator molecules employed anti-CD80 (16-10A1), anti-CD86 (GL1),
and anti-CD40 (FGK45.5), used as FITC conjugates. Anti-DEC-205
(NLDC-145) and anti-CD11b (M-170) were used as FITC conjugates.
Propidium iodide (PI; 1 µg/mL) was included in the final wash after
immunofluorescent staining to label dead cells.
Flow cytometric analysis and sorting of DCs Most analyses were done on a fluorescence-activated cell-sorter scanner (FACStar Plus; Becton Dickinson, San Jose, CA), as described previously,4 by using up to 4 fluorescent channels for the immunofluorescent staining (FL1 for FITC, FL2 for PE, FL3 for Cy5, and FL4 for Alexa 594) with the FL5 channel set to exclude PI-positive dead cells and autofluorescent cells. During gating, care was taken to ensure that any cells brightly fluorescent in FL3 and spilling over into FL5 were not gated out as dead cells. A MoFlo instrument (Cytomation, Fort Collins, CO) was used to sort DC populations. To sort or gate for DCs, the class II MHC and CD11c markers, together with the forward and side scatter gates, were set to select for cells with DC characteristics. The expression of CD4 and CD8 was used to segregate DC subpopulations (CD8+CD4 , CD8 intermediate
[int] CD4, CD8CD4, and
CD4+CD8). Reanalysis of sorted DC populations
was done, and only DCs with a purity of greater than 95% were used for
further functional analyses.
Stimulation of DCs for cytokine production Sorted splenic mouse DCs (0.5-2 × 106/mL) were cultured in 96-well round-bottomed plates in modified RPMI-1640 medium containing 10% fetal-calf serum (FCS) at 37°C in an atmosphere of 10% carbon dioxide (CO2) in air, as described previously.25 The culture supernatant was collected, separated from the cells by centrifugation, and stored at 20°C
until analysis. Conditions previously established for optimum
production of each cytokine were used.11 For production of
IL-12, the stimulation mixture was GM-CSF (200 U/mL), IFN- (20 ng/mL), CpG-ph (0.5 µM), or SAC (10 µg Pansorbin/mL) in the presence or absence of IL-4 (100 U/mL). Incubation time was 16 to 24 hours. For production of IFN-, IL-12 and IL-18 (20 ng/mL each) were
used for stimulation and the incubation time was 3 or 4 days.
Analysis of IL-12 and IFN- ; ATCC). Cytokine binding was then
detected with an appropriate biotinylated detection mAb, namely R1-5D9
(anti-IL-12 p40; ATCC), C17.8 (anti-IL-12 p40; hybridoma provided by
L. Schofield, WEHI), or XMG1.2 (anti-IFN-; ATCC).
Allostimulatory mixed leukocyte reaction CD4+ and CD8+ T cells were purified from pooled mesenteric, axillary, brachial, and inguinal lymph nodes of CBA/J mice. Briefly, cell suspensions from lymph nodes were obtained by passing the organs through a fine mesh sieve. Cells were washed with PBS containing 2% FCS. The cells were then depleted of cells other than T cells by incubation with a mixture of the following antibodies: Ter119, RA3-6B2, M1-70, RB6-6CS, and either anti-CD8 (53.6.7) or anti-CD4 (GK1.5). Antibody-coated cells were removed by incubation with sheep anti-rat IgG magnetic beads (Dynal). The CD4+ or CD8+ T cells (98% and 96% pure, respectively) were washed, suspended in culture medium (modified RPMI-164025 containing 10% FCS), and counted. Replicate culture trays were incubated at 37°C in 10% CO2 in air for 2.5 to 5.5 days. On days 2.5, 3.5, 4.5, and 5.5, a culture tray was pulsed with tritium-thymidine (1 µCi [0.0185 MBq]/well) for 6 hours and then frozen. The trays were thawed, the cells were harvested on glass fiber filters, and the amount of thymidine incorporated was measured by using liquid scintillation. All cultures (all time points) were done in triplicate or greater, and background controls with T cells only were included for each time point.
Dose response of ProGP-1 One aim of this study was to compare the effect on DC expansion of ProGP-1, an agonist of the Flt-3 and G-CSF receptors, with that of the individual cytokines FL, G-CSF, and pGM-CSF. The optimal dose of injected FL (10 µg/day for 10 days) and pGM-CSF (2 µg/day for 5 days)14,17,18 for normal enhancement of DCs in mouse spleens was previously determined by others, and these regimens also worked well in our laboratory. To establish the optimal dose and duration of ProGP-1 administration, mice were injected with 10, 20, 50, or 100 µg ProGP-1/day for either 7 or 10 days, and splenic DCs were isolated and counted. The optimal regimen was found to be 20 µg/day for 10 days, which increased DC numbers to about 5 × 107 extracted DCs/spleen. Higher doses of ProGP-1 provided similar or occasionally higher DC yields but were associated with complications caused by excessive hematopoiesis. For example, some mice had such marked expansion of vertebral bone marrow that hematopoietic cells encroached on regional peripheral nerves, nerve roots, and the spinal cord. In some of these mice, paresis or paralysis developed. All mice given ProGP-1 had enlarged spleens (weight increase ~5-fold) and lymph nodes. Serum samples from these mice were examined for anti-ProGP-1 antibodies by ELISA (data not shown). Mice that received 1 course of treatment (10 days) or 2 courses of treatment (10 days each with a 2-week break between them) had anti-ProGp-1 serum titers that were 2-fold higher than those of untreated controls (mean titer, 1:512 versus 1:256; 5 mice/group). Moreover, mice that received 2 courses of ProGP-1 treatment had a normal response to the cytokine treatment. Thus, there was little evidence suggesting that the ProGP-1 chimeric cytokine was itself immunogenic in vivo.Effect of ProGP-1 treatment on mouse spleen DCs To investigate the effect of ProGP-1 on murine DCs, groups of mice were injected with 20 µg ProGP-1/day for 10 days and splenic DCs were isolated and analyzed for various surface markers by flow cytometry. The results were compared with those in untreated control mice and in mice given both FL and G-CSF (10 µg of each/day); muFL was used initially to provide the best control for effects on DCs in murine spleen. The total numbers of splenic DCs of the main subtypes after these treatments are shown in Table 1. Results of the flow cytometric analysis of the surface phenotypes of the DCs after treatment with ProGP-1 are provided in Figure 1.
ProGP-1 treatment increased the total DCs per spleen about 30 fold. This increase was heavily biased toward CD8+ DCs,
which increased 130 fold. The CD8+ DC population was
composed of CD8-high (hi) cells, as well as many CD8int
cells. The CD8int DCs could be separated as a distinct
population not only by their level of CD8 expression but also by their
forward-scatter profile, which was intermediate between that of
CD4 Kinetics of DC response to ProGP-1 We previously showed that the 3 main splenic DC subsets (CD4+CD8, CD4CD8,
and CD4CD8+) have different turnover rates
and independent developmental kinetics.9 To determine
whether the selective enhancement of CD8+ DCs after ProGP-1
treatment occurred throughout the response, DCs from mouse spleens were
isolated, enumerated, and analyzed throughout the 16 days of ProGP-1
treatment (Figure 2). We found that the
DCs in each subset had increased in size, as assessed by forward light
scatter, as early as 12 hours after ProGP-1 injection (data not shown).
Moreover, as early as 2.5 days after ProGP-1 treatment began, the
CD8+ DCs had become the principal subset in the spleen. By
day 4.5 of treatment, the number of CD8hi DCs had increased
7 fold and the number of CD8int DCs had increased 11 fold.
CD8+ DCs then remained the dominant subtype, showing an
increase of about 130 fold at the day-10 peak. In contrast,
CD4CD8 DCs and
CD4+CD8 DCs had maximal increases of 8 fold
and 2.5 fold, respectively, on day 8.5. After 16 days of ProGP-1
treatment, the numbers of CD8 DCs were close to those in
untreated mice. The numbers of CD8+ DCs were still elevated
(by about 16 fold), although the maximal effects of ProGP-1 on
CD8+ DC numbers were observed on about day 10 of treatment.
The surface markers shown in Figure 1 remained unchanged throughout the
treatment (data not shown).
Effect of rmuFL and rhuFL on mouse spleen DCs To provide a direct comparison with ProGP-1, the effect of FL alone on DCs in mouse spleens was determined. Initially, an rmuFL preparation derived from a mammalian cell line was used because we wanted to compare the effect of ProGP-1 with that of the normal ligand of the murine Flt-3 receptor. As shown in Table 1 and Figure 3, muFL treatment increased DC numbers about 11 fold, and this increase was heavily biased toward CD8+ DCs, which increased 32 fold. Thus, the effect of FL was similar in type, though not in magnitude, to that of ProGP-1.
This result, however, differed from findings in previous studies in
which a more extensive and more balanced enhancement of both the
CD8 Effect of G-CSF and pGM-CSF on mouse spleen DCs The effects on splenic DCs of 2 myeloid cytokines, G-CSF and pGM-CSF, were compared with the effects of FL and ProGP-1. Treatment with G-CSF yielded a small increase (2.3 fold) in total DC numbers, with a small bias (3-fold increase) toward CD8+ DCs (Table 1 and Figure 4). Apart from typical CD11chi MHC class IIhi DCs, G-CSF treatment resulted in a large population of cells that were CD11cintCD8int and positive for MHC class II; because it was not clear that these were bona fide DCs, however, they are not included in Table 1 or Figure 4.
Treatment with pGM-CSF provided the greatest contrast with the
effects of all the other cytokines tested, and the results were similar
to those of a previous study.18 Previously, it was not
known whether pGM-CSF differentially affected the
CD4+CD8 Maturation of cytokine-treated DCs in culture To determine whether the DCs from mice given cytokines could be matured further in culture and whether one subset could be transformed into another, sorted DCs from mice treated with ProGP-1, muFL, both muFL and G-CSF, and pGM-CSF were stained and analyzed after overnight culture in medium alone or with various cytokines and stimuli. DCs from cytokine-treated mice behaved in a manner similar to that of the corresponding DC population from normal mice.9 Specifically, all subpopulations could be matured further by culture in medium alone (as indicated by increases in class II MHC expression and forward light scatter), and such maturation could be enhanced further by adding GM-CSF and factors such as tumor necrosis factor (TNF-) or CpG-ph. With all cytokine and stimulus combinations tested
(including GM-CSF together with IFN-, TNF-, IL-1, IL-3, FL,
ProGP-1, CpG-ph, and various combinations of these agents), the CD4 and
CD8 phenotype of the DCs from treated or control mice did not change
except for a decrease in the level of CD4 expression. This confirmed
earlier results with DCs from untreated mice9 indicating
that the DC populations did not transform into other populations under
conditions of maturation or activation. Moreover, the new group of
CD8int DCs identified in the treated mice remained
CD8int after overnight culture, after 60 hours of culture,
and under all conditions tested, despite the further maturation of
these DCs indicated by increases in their size and MHC class II expression.
IL-12 production by DCs from cytokine-treated mice CD8+ splenic DCs in normal mice are by far the greatest producers of all isoforms of IL-12, including the bioactive p70 form, the p40 monomer, and the receptor antagonist (p40)2 form.11,17,26-29 Optimal production of the p70 form of IL-12 requires a stimulus of bacterial, microbial, or T-cell-dependent origin and an appropriate mixture of cytokines, including IL-4.30,31 Under the same conditions but in the absence of IL-4, production of p40 most notably the (p40)2
antagonist formis favored. Because CD8+ DCs were
increased 130 fold with ProGP-1 and more than 30 fold with muFL in our
experiments, we conducted studies to establish whether these
cytokine-enhanced CD8+ DCs were indeed capable of high
levels of IL-12 production. We also examined whether the ratio of total
p40 to p70 was similarly regulated by IL-4 in the treated DCs.
Therefore, all the sorted DC subpopulations from cytokine-treated mice
were stimulated in culture with CpG-ph in the presence of GM-CSF and
IFN- and with or without IL-4 (data not shown), and the production
of IL-12 p70 and p40 in the culture supernatants was determined by
ELISA (Figure 5). In addition, the
production of p40, p70, and (p40)2 was assessed by Western
blotting (data not shown).
The highly abundant CD8+ DCs from mice treated with ProGP-1, muFL, and both muFL and G-CSF all produced high per-cell levels of p70 (maximized in the presence of IL-4) and p40, similar to results with CD8+ DCs from control mice. The CD8int DCs from these mice produced levels of IL-12 resembling those of the CD8hi DCs (Figure 5). IL-12 production of CD8+ DCs from G-CSF-treated mice was about 70% of control values. However, the minority population of CD8+ DCs and CD8int DCs from mice treated with pGM-CSF produced only small amounts of IL-12 p70 but near normal levels of p40 and, in line with this finding, Western blot analysis showed a predominance of the antagonistic (p40)2 form, even in the presence of IL-4 (data not shown). The CD4+CD8 Splenic DCs from mice treated with ProGP-1, muFL, and both muFL and G-CSF all followed the same pattern of IL-12 regulation observed in normal mice (Figure 5), since when IL-4 was omitted, p70 production dropped, p40 production increased, and levels of the antagonistic (p40)2 form increased (data not shown). However, this tight control was abrogated in splenic DCs from pGM-CSF-treated mice, since even in the presence of IL-4, there was a negligible amount of p70 and a predominance of p40 and (p40)2. This regulation and production of IL-12 by each of the DC populations from cytokine-treated mice followed the same pattern when other stimuli, such as SAC or CD40L, were used (data not shown). Most notably, as was the case with DCs from untreated mice, none of the DC populations from treated mice produced appreciable levels of bioactive IL-12 in the absence of a microbial or T-cell-derived stimulus. IFN- CD8 DCs of normal mice have the greatest
potential to produce IFN-, as revealed by ELISA assay of culture
supernatants after stimulation with a combination of IL-12 and
IL-18.11 The capacity of the
CD4CD8 DCs from the cytokine-treated mice
to produce IFN- is illustrated in Figure
6. Only the
CD4CD8 DCs from mice treated with G-CSF
produced levels of IFN- comparable to control values; the
CD4CD8 DCs from mice treated with ProGP-1,
muFL, both muFL and GM-CSF, and pGM-CSF all showed severe defects in
the production of IFN-. The CD4+CD8 and
CD4CD8+ DC populations all produced small
amounts of IFN-less than half of that produced by the equivalent
DC subsets of normal mice (data not shown). The defect in IFN-
production in ProGP-1-treated mice was manifest as early as 2 days
after treatment began and was extensive at 5 days; at 10 days,
production of IFN- was barely detectable (Figure 6).
Allostimulatory capacity of DCs from cytokine-treated mice Because an exceptional capacity to stimulate T cells in an allogenic mixed leukocyte reaction (MLR) is a hallmark of mature DCs, we tested whether the DC subpopulations from cytokine-treated mice maintained this capacity. All DC subpopulations in normal mouse spleens have a strong capacity to activate naive allogenic CD4 or CD8 T cells to cycle, and we found this also to be true of DC populations from treated mice. The main DC populations (CD4+CD8, CD4CD8,
and CD4CD8+ DCs) from mice treated with
ProGP-1, muFL, and G-CSF for 10 days were compared with those from
normal mice (Figure 7). There was little
difference between the DCs from treated mice and those from normal mice
in their ability to stimulate naive CD4 T cells; thus, this aspect of
antigen presentation was intact in the cytokine-treated mice. The
allostimulatory capacity of DCs from pGM-CSF-treated mice was
previously studied by others17 and also found to be intact.
We also assessed the CD8int DC population that was a minor
component of normal spleens but was enhanced in spleens from
cytokine-treated mice, since these DCs might have represented immature
forms of CD8hi DC. With care taken to eliminate
autofluorescent macrophages,4 we obtained a small sample
of these CD8int DCs from normal spleens and found their
stimulatory capacity to be similar to that of
CD4
Of all the cytokines tested in this study, ProGP-1, an agonist of
the Flt-3 and G-CSF receptors, was the most effective in increasing the
number of spleen DCs. It was substantially more effective than FL or
G-CSF used alone or together. However, its effects were strongly
selective, increasing the number of CD4 One surprising finding was the difference between results from
administration to mice of rmuFL prepared in mammalian cells and
administration of rhuFL. This difference partly accounts for why a
strong selective effect of FL was not observed in an earlier study
using huFL.14 The huFL produced equally large increases in
CD8 The exact reason for the different effects of the muFL and huFL preparations remains unclear. It may be simply that different recombinant expression systems produce differentially glycosylated or folded FL proteins with large differences in blood life span or other bioactivities. However, it does appear that there is a qualitative as well as a quantitative difference between the preparations that may reflect a species-specific difference between them. FL is structurally related to other members of the tyrosine kinase receptor family,33 and possibly huFL is "promiscuous" in the murine system and able to bind to other, related members of this family. Alternatively, differences in affinity of binding to the Flt-3 receptor may lead to differences in signaling and consequential downstream biologic activities. Several signaling pathways and molecules found to be important in Flt-3 signaling34-37 may be differentially activated by muFL and huFL. Only studies of the kinetics and affinity of binding to Flt-3 receptors and downstream intracellular signaling events would clarify this issue. One "new" splenic DC subtype emphasized by this study
consists of DCs bearing intermediate levels of CD8 A consequence of the marked enhancement of CD8+ DCs with
ProGP-1 or FL treatment is potentially high levels of IL-12-producing DCs with a capacity to bias T-cell immune responses to the T-helper-1 (Th1) type. It was therefore important to determine whether the expanded CD8+ DCs had the same capacity to produce IL-12
and if the process was under normal regulation. We found the capacity
to produce high levels of IL-12 p70 and p40 was indeed maintained and
restricted to CD8+ DCs from mice treated with ProGP-1, FL,
and G-CSF. The IL-12 regulatory systems also seemed intact. This result
has important implications for clinical use of these cytokines in
immune modulation, since treatment with these agents could lead Despite the functional and phenotypic normality of most of the expanded
DC populations, a marked deficit was found in their ability to make
IFN- Overall, this study found that ProGP-1, along with FL, G-CSF, and
pGM-CSF, are useful tools for expanding DC populations. However, the
expansion is often strongly selective for particular DC subpopulations
and this could have important biologic and clinical implications. Many
of the biologic functions of the expanded DCs remain intact, but
some
We thank V. Lapatis, D. Kaminaris, J. Chan, C. Tarlinton, G. Paukovics, and F. Battye for excellent FACS assistance.
Submitted March 8, 2001; accepted October 30, 2001.
Supported in part by research funding from Pharmacia, St Louis, MO (K.S.). H.H. was supported by a Deutsche Krebshilfe fellowship.
Two of the authors (R.E. and S.W.) are employed by Pharmacia Corporation, St Louis, MO.
The publication costs of this article were defrayed in part by page charge payment. Therefore, and solely to indicate this fact, this article is hereby marked "advertisement" in accordance with 18 U.S.C. section 1734.
Reprints: Meredith O'Keeffe, Walter and Eliza Hall Institute of Medical Research, Royal Melbourne Hospital, Victoria, 3050, Australia.
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  Z. Shao, T. O. Makinde, H. S. McGee, X. Wang, and D. K. Agrawal Fms-Like Tyrosine Kinase 3 Ligand Regulates Migratory Pattern and Antigen Uptake of Lung Dendritic Cell Subsets in a Murine Model of Allergic Airway Inflammation J. Immunol., December 1, 2009; 183(11): 7531 - 7538. [Abstract] [Full Text] [PDF]
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 B. B. Ganesh, D. M. Cheatem, J. R. Sheng, C. Vasu, and B. S. Prabhakar GM-CSF-induced CD11c+CD8a--dendritic cells facilitate Foxp3+ and IL-10+ regulatory T cell expansion resulting in suppression of autoimmune thyroiditis Int. Immunol., March 1, 2009; 21(3): 269 - 282. [Abstract] [Full Text] [PDF]
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 K. R. Mott and H. Ghiasi Role of Dendritic Cells in Enhancement of Herpes Simplex Virus Type 1 Latency and Reactivation in Vaccinated Mice Clin. Vaccine Immunol., December 1, 2008; 15(12): 1859 - 1867. [Abstract] [Full Text] [PDF]
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 K. R. Mott, D. UnderHill, S. L. Wechsler, and H. Ghiasi Lymphoid-Related CD11c+ CD8{alpha}+ Dendritic Cells Are Involved in Enhancing Herpes Simplex Virus Type 1 Latency J. Virol., October 15, 2008; 82(20): 9870 - 9879. [Abstract] [Full Text] [PDF]
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E. Carreras, S. Turner, V. Paharkova-Vatchkova, A. Mao, C. Dascher, and S. Kovats Estradiol Acts Directly on Bone Marrow Myeloid Progenitors to Differentially Regulate GM-CSF or Flt3 Ligand-Mediated Dendritic Cell Differentiation J. Immunol., January 15, 2008; 180(2): 727 - 738. [Abstract] [Full Text] [PDF]
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S. Takenaka, E. Safroneeva, Z. Xing, and J. Gauldie Dendritic Cells Derived from Murine Colonic Mucosa Have Unique Functional and Phenotypic Characteristics J. Immunol., June 15, 2007; 178(12): 7984 - 7993. [Abstract] [Full Text] [PDF]
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D. Yadav and N. Sarvetnick B7-2 Regulates Survival, Phenotype, and Function of APCs J. Immunol., May 15, 2007; 178(10): 6236 - 6241. [Abstract] [Full Text] [PDF]
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Y. S. Kim, S. H. Yang, H. G. Kang, E. Y. Seong, S. H. Lee, W. Gao, J. Kenny, X. X. Zheng, and T. B. Strom Distinctive role of donor strain immature dendritic cells in the creation of allograft tolerance Int. Immunol., December 1, 2006; 18(12): 1771 - 1777. [Abstract] [Full Text] [PDF]
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J. R. Sheng, L. Li, B. B. Ganesh, C. Vasu, B. S. Prabhakar, and M. N. Meriggioli Suppression of Experimental Autoimmune Myasthenia Gravis by Granulocyte-Macrophage Colony-Stimulating Factor Is Associated with an Expansion of FoxP3+ Regulatory T Cells J. Immunol., October 15, 2006; 177(8): 5296 - 5306. [Abstract] [Full Text] [PDF]
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H. Ito, E. Esashi, T. Akiyama, J.-i. Inoue, and A. Miyajima IL-18 produced by thymic epithelial cells induces development of dendritic cells with CD11b in the fetal thymus Int. Immunol., August 1, 2006; 18(8): 1253 - 1263. [Abstract] [Full Text] [PDF]
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M. O'Keeffe, R. J. Grumont, H. Hochrein, M. Fuchsberger, R. Gugasyan, D. Vremec, K. Shortman, and S. Gerondakis Distinct roles for the NF-{kappa}B1 and c-Rel transcription factors in the differentiation and survival of plasmacytoid and conventional dendritic cells activated by TLR-9 signals Blood, November 15, 2005; 106(10): 3457 - 3464. [Abstract] [Full Text] [PDF]
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R. Tussiwand, N. Onai, L. Mazzucchelli, and M. G. Manz Inhibition of Natural Type I IFN-Producing and Dendritic Cell Development by a Small Molecule Receptor Tyrosine Kinase Inhibitor with Flt3 Affinity J. Immunol., September 15, 2005; 175(6): 3674 - 3680. [Abstract] [Full Text] [PDF]
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K. P. A. MacDonald, V. Rowe, H. M. Bofinger, R. Thomas, T. Sasmono, D. A. Hume, and G. R. Hill The Colony-Stimulating Factor 1 Receptor Is Expressed on Dendritic Cells during Differentiation and Regulates Their Expansion J. Immunol., August 1, 2005; 175(3): 1399 - 1405. [Abstract] [Full Text] [PDF]
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S. H. Naik, A. I. Proietto, N. S. Wilson, A. Dakic, P. Schnorrer, M. Fuchsberger, M. H. Lahoud, M. O'Keeffe, Q.-x. Shao, W.-f. Chen, et al. Cutting Edge: Generation of Splenic CD8+ and CD8- Dendritic Cell Equivalents in Fms-Like Tyrosine Kinase 3 Ligand Bone Marrow Cultures J. Immunol., June 1, 2005; 174(11): 6592 - 6597. [Abstract] [Full Text] [PDF]
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M. O'Keeffe, T. C. Brodnicki, B. Fancke, D. Vremec, G. Morahan, E. Maraskovsky, R. Steptoe, L. C. Harrison, and K. Shortman Fms-like tyrosine kinase 3 ligand administration overcomes a genetically determined dendritic cell deficiency in NOD mice and protects against diabetes development Int. Immunol., March 1, 2005; 17(3): 307 - 314. [Abstract] [Full Text] [PDF]
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T. Tamura, P. Tailor, K. Yamaoka, H. J. Kong, H. Tsujimura, J. J. O'Shea, H. Singh, and K. Ozato IFN Regulatory Factor-4 and -8 Govern Dendritic Cell Subset Development and Their Functional Diversity J. Immunol., March 1, 2005; 174(5): 2573 - 2581. [Abstract] [Full Text] [PDF]
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K. P. A. MacDonald, V. Rowe, A. D. Clouston, J. K. Welply, R. D. Kuns, J. L. M. Ferrara, R. Thomas, and G. R. Hill Cytokine Expanded Myeloid Precursors Function as Regulatory Antigen-Presenting Cells and Promote Tolerance through IL-10-Producing Regulatory T Cells J. Immunol., February 15, 2005; 174(4): 1841 - 1850. [Abstract] [Full Text] [PDF]
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K. McKenna, A.-S. Beignon, and N. Bhardwaj Plasmacytoid Dendritic Cells: Linking Innate and Adaptive Immunity J. Virol., January 1, 2005; 79(1): 17 - 27. [Full Text] [PDF]
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N. Teleshova, J. Jones, J. Kenney, J. Purcell, R. Bohm, A. Gettie, and M. Pope Short-term Flt3L treatment effectively mobilizes functional macaque dendritic cells J. Leukoc. Biol., June 1, 2004; 75(6): 1102 - 1110. [Abstract] [Full Text] [PDF]
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C. Johansson and M. J. Wick Liver Dendritic Cells Present Bacterial Antigens and Produce Cytokines upon Salmonella Encounter J. Immunol., February 15, 2004; 172(4): 2496 - 2503. [Abstract] [Full Text] [PDF]
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T. Kumamoto, D. Shalhevet, H. Matsue, M. E. Mummert, B. R. Ward, J. V. Jester, and A. Takashima Hair follicles serve as local reservoirs of skin mast cell precursors Blood, September 1, 2003; 102(5): 1654 - 1660. [Abstract] [Full Text] [PDF]
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 A. D'Amico and L. Wu The Early Progenitors of Mouse Dendritic Cells and Plasmacytoid Predendritic Cells Are within the Bone Marrow Hemopoietic Precursors Expressing Flt3 J. Exp. Med., July 21, 2003; 198(2): 293 - 303. [Abstract] [Full Text] [PDF]
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H. Karsunky, M. Merad, A. Cozzio, I. L. Weissman, and M. G. Manz Flt3 Ligand Regulates Dendritic Cell Development from Flt3+ Lymphoid and Myeloid-committed Progenitors to Flt3+ Dendritic Cells In Vivo J. Exp. Med., July 21, 2003; 198(2): 305 - 313. [Abstract] [Full Text] [PDF]
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S. Naik, D. Vremec, L. Wu, M. O'Keeffe, and K. Shortman CD8{alpha}+ mouse spleen dendritic cells do not originate from the CD8{alpha}- dendritic cell subset Blood, July 15, 2003; 102(2): 601 - 604. [Abstract] [Full Text] [PDF]
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C. Vasu, R.-N. E. Dogan, M. J. Holterman, and B. S. Prabhakar Selective Induction of Dendritic Cells Using Granulocyte Macrophage-Colony Stimulating Factor, But Not fms-Like Tyrosine Kinase Receptor 3-Ligand, Activates Thyroglobulin-Specific CD4+/CD25+ T Cells and Suppresses Experimental Autoimmune Thyroiditis J. Immunol., June 1, 2003; 170(11): 5511 - 5522. [Abstract] [Full Text] [PDF]
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 V. Pullarkat, P. P. Lee, R. Scotland, V. Rubio, S. Groshen, C. Gee, R. Lau, J. Snively, S. Sian, S. L. Woulfe, et al. A Phase I Trial of SD-9427 (Progenipoietin) with a Multipeptide Vaccine for Resected Metastatic Melanoma Clin. Cancer Res., April 1, 2003; 9(4): 1301 - 1312. [Abstract] [Full Text] [PDF]
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G. Miller, V. G. Pillarisetty, A. B. Shah, S. Lahrs, and R. P. DeMatteo Murine Flt3 Ligand Expands Distinct Dendritic Cells with Both Tolerogenic and Immunogenic Properties J. Immunol., April 1, 2003; 170(7): 3554 - 3564. [Abstract] [Full Text] [PDF]
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G. Sailaja, S. Husain, B. P. Nayak, and A. M. Jabbar Long-Term Maintenance of gp120-Specific Immune Responses by Genetic Vaccination with the HIV-1 Envelope Genes Linked to the Gene Encoding Flt-3 Ligand J. Immunol., March 1, 2003; 170(5): 2496 - 2507. [Abstract] [Full Text] [PDF]
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K. P. A. MacDonald, V. Rowe, C. Filippich, R. Thomas, A. D. Clouston, J. K. Welply, D. N. J. Hart, J. L. M. Ferrara, and G. R. Hill Donor pretreatment with progenipoietin-1 is superior to granulocyte colony-stimulating factor in preventing graft-versus-host disease after allogeneic stem cell transplantation Blood, March 1, 2003; 101(5): 2033 - 2042. [Abstract] [Full Text] [PDF]
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M. O'Keeffe, H. Hochrein, D. Vremec, B. Scott, P. Hertzog, L. Tatarczuch, and K. Shortman Dendritic cell precursor populations of mouse blood: identification of the murine homologues of human blood plasmacytoid pre-DC2 and CD11c+ DC1 precursors Blood, February 15, 2003; 101(4): 1453 - 1459. [Abstract] [Full Text] [PDF]
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G. Schiavoni, F. Mattei, P. Sestili, P. Borghi, M. Venditti, H. C. Morse III, F. Belardelli, and L. Gabriele ICSBP Is Essential for the Development of Mouse Type I Interferon-producing Cells and for the Generation and Activation of CD8{alpha}+ Dendritic Cells J. Exp. Med., December 2, 2002; 196(11): 1415 - 1425. [Abstract] [Full Text] [PDF]
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M. O'Keeffe, H. Hochrein, D. Vremec, I. Caminschi, J. L. Miller, E. M. Anders, L. Wu, M. H. Lahoud, S. Henri, B. Scott, et al. Mouse Plasmacytoid Cells: Long-lived Cells, Heterogeneous in Surface Phenotype and Function, that Differentiate Into CD8+ Dendritic Cells Only after Microbial Stimulus J. Exp. Med., November 18, 2002; 196(10): 1307 - 1319. [Abstract] [Full Text] [PDF]
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G. Miller, V. G. Pillarisetty, A. B. Shah, S. Lahrs, Z. Xing, and R. P. DeMatteo Endogenous Granulocyte-Macrophage Colony-Stimulating Factor Overexpression In Vivo Results in the Long-Term Recruitment of a Distinct Dendritic Cell Population with Enhanced Immunostimulatory Function J. Immunol., September 15, 2002; 169(6): 2875 - 2885. [Abstract] [Full Text] [PDF]
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Abstract
Introduction
