A Discrete Subset of Monocyte-Derived Cells among Typical Conventional Type 2 Dendritic Cells Can Efficiently Cross-Present

AbstractCyclic AMP (cAMP) is involved in multiple biological processes. However, little is known about its role in shaping immunity. Here we show that cAMP-PKA-CREB signaling (a pattern recognition receptor [PRR]-independent) regulates conventional type-2 Dendritic Cells (cDC2s), but not cDC1s and reprograms their Th17-inducing properties via repression of IRF4 and KLF4, transcription factors (TFs) for Th2 induction. Genetic loss of IRF4 phenocopies the effects of cAMP signaling on Th17-induction, indicating that the cAMP effect is secondary to repression of IRF4. Moreover, signaling in cDC2s by a PRR-dependent microbial product, curdlan, represses IRF4 and KLF4, resulting in a pro-Th17 phenotype. These results define a novel signaling pathway by which cDC2s display plasticity and provide a new molecular basis for the novel cDC2 and cDC17 classification. In addition, the data reveal that cAMP signaling can alter DCs function and fate by repressing IRF4 and KLF4, a pathway that can be harnessed for immuno-regulation.

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Inhibition of IRF4 in dendritic cells by PRR-independent and -dependent signals inhibit Th2 and promote Th17 responses

eLife ◽

10.7554/elife.49416 ◽

2020 ◽

Vol 9 ◽

Cited By ~ 1

Author(s):

Jihyung Lee ◽

Junyan Zhang ◽

Young-Jun Chung ◽

Jun Hwan Kim ◽

Chae Min Kook ◽

...

Keyword(s):

Dendritic Cells ◽

Biological Processes ◽

Microbial Product ◽

Independent Mechanism ◽

Conventional Type ◽

Recognition Receptor

Cyclic AMP (cAMP) is involved in many biological processes but little is known regarding its role in shaping immunity. Here we show that cAMP-PKA-CREB signaling (a pattern recognition receptor [PRR]-independent mechanism) regulates conventional type-2 Dendritic Cells (cDC2s) in mice and reprograms their Th17-inducing properties via repression of IRF4 and KLF4, transcription factors essential for cDC2-mediated Th2 induction. In mice, genetic loss of IRF4 phenocopies the effects of cAMP on Th17 induction and restoration of IRF4 prevents the cAMP effect. Moreover, curdlan, a PRR-dependent microbial product, activates CREB and represses IRF4 and KLF4, resulting in a pro-Th17 phenotype of cDC2s. These in vitro and in vivo results define a novel signaling pathway by which cDC2s display plasticity and provide a new molecular basis for the classification of novel cDC2 and cDC17 subsets. The findings also reveal that repressing IRF4 and KLF4 pathway can be harnessed for immuno-regulation.

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Myocarditis Elicits Dendritic Cell and Monocyte Infiltration in the Heart and Self-Antigen Presentation by Conventional Type 2 Dendritic Cells

Frontiers in Immunology ◽

10.3389/fimmu.2018.02714 ◽

2018 ◽

Vol 9 ◽

Cited By ~ 7

Author(s):

Katrien Van der Borght ◽

Charlotte L. Scott ◽

Liesbet Martens ◽

Dorine Sichien ◽

Gert Van Isterdael ◽

...

Keyword(s):

Dendritic Cells ◽

Dendritic Cell ◽

Antigen Presentation ◽

Self Antigen ◽

Conventional Type

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STAT6 signaling pathway controls germinal center responses promoted after antigen targeting to conventional type 2 dendritic cells

Current Research in Immunology ◽

10.1016/j.crimmu.2021.08.001 ◽

2021 ◽

Vol 2 ◽

pp. 120-131

Author(s):

Fernando Bandeira Sulczewski ◽

Larissa Alves Martino ◽

Bianca da Silva Almeida ◽

Márcio Massao Yamamoto ◽

Daniela Santoro Rosa ◽

...

Keyword(s):

Dendritic Cells ◽

Signaling Pathway ◽

Germinal Center ◽

Conventional Type

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Select hyperactivating NLRP3 ligands enhance the TH1- and TH17-inducing potential of human type 2 conventional dendritic cells

Science Signaling ◽

10.1126/scisignal.abe1757 ◽

2021 ◽

Vol 14 (680) ◽

pp. eabe1757

Author(s):

Lukas Hatscher ◽

Christian H. K. Lehmann ◽

Ariawan Purbojo ◽

Constantin Onderka ◽

Chunguang Liang ◽

...

Keyword(s):

Dendritic Cells ◽

Immune Responses ◽

Vaccine Development ◽

Inflammatory Diseases ◽

Toll Like Receptor ◽

T Helper ◽

Conventional Type ◽

Prime Target

The detection of microorganisms and danger signals by pattern recognition receptors on dendritic cells (DCs) and the consequent formation of inflammasomes are pivotal for initiating protective immune responses. Although the activation of inflammasomes leading to secretion of the cytokine IL-1β is typically accompanied by pyroptosis (an inflammatory form of lytic programmed cell death), some cells can survive and exist in a state of hyperactivation. Here, we found that the conventional type 2 DC (cDC2) subset is the major human DC subset that is transcriptionally and functionally poised for inflammasome formation and response without pyroptosis. When cDC2 were stimulated with ligands that relatively weakly activated the inflammasome, the cells did not enter pyroptosis but instead secreted IL-12 family cytokines and IL-1β. These cytokines induced prominent T helper type 1 (TH1) and TH17 responses that were superior to those seen in response to Toll-like receptor (TLR) stimulation alone or to stronger, classical inflammasome ligands. These findings not only define the human cDC2 subpopulation as a prime target for the treatment of inflammasome-dependent inflammatory diseases but may also inform new approaches for adjuvant and vaccine development.

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EFFECT OF DNA CONSTRUCTIONS ELECTROPORATION ON DENDRITIC CELLS

Medical Immunology (Russia) ◽

10.15789/1563-0625-eod-2243 ◽

2021 ◽

Vol 23 (4) ◽

pp. 653-658

Author(s):

A. S. Bulygin ◽

V. P. Tereshchenko ◽

R. Yu. Zavodskii ◽

I. A. Obleukhova ◽

S. V. Sennikov ◽

...

Keyword(s):

Dendritic Cells ◽

Cell Cultures ◽

Culture Medium ◽

Bone Marrow Cells ◽

Culture Media ◽

Pcr Analysis ◽

Electric Impulse ◽

Plasmacytoid Dcs ◽

Conventional Type

Today transfection of mammalian cell with DNA or RNA construction is the only method for delivering programmed information into the cell nucleus. Electroporation is most commonly used method of transfection in experiments with dendritic cell. The aim of electroporation is to permeabilize the membrane by passing electric impulse through the cell. Due to the increase permeability of the membrane chance DNA or RNA construction getting inside into the cell is increased, wherein survival of the cells is decreased.In the study male mice C57Bl/6 line 2-4 months old were used. From femur bones was isolated 20 × 106 bone marrow cells, which were cultured in 20 mL of complete RPMI-1640 for 7 days. To generate dendritic cells from BM cells, 100 ng/mL of rmFlt3-L was added to culture media at day 0. After 7 days of cultivation, the cell cultures were electroporated with control noncoding plasmids p5 (EP P5) or pmaxCCR9 encoding mouse chemokine receptor CCR9 (EP CCR9). The controls were cell cultures electroporated without any plasmids (mock EP) and cell cultures without electroporation (none EP). 5 × 105 cells were electroporated and resting for 10 minutes. After 10 minutes, cells were harvested and seeded into 24-well plates in 1 mL of culture medium and conditioning medium (1:1). Then, 50 ng/mL of Flt3-L was added to each well. The next day, transfected cells were collected and used for flow cytometry, qRT-PCR analysis.It was found that after electroporation in the groups mock EP, EP P5, EP CCR9 relative amount of live CD11c+ dendritic cells was significantly less than in the non EP group. Moreover, in the EP P5 and EP CCR9 groups the frequency of live CD11c+ dendritic cells was significantly less than in the mock EP group. Expression of the CD86 marker in the EP P5 and EP CCR9 groups was significantly higher than in the non EP and mock EP groups. Expression of the I-AB(MHCII) in the EP CCR9 group on cDC2s was significantly higher than in the non EP group. On plasmacytoid DCs (pDCs) and conventional type 2 DCs (cDC2s) in the EP CCR9 group expression of CCR9 was significantly higher than in the non EP group. Therefore, in this study, we demonstrated the effectiveness of electroporation, accompanied by the decrease in the survival rate and maturation of DCs.

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