Publications by authors named "Theresa L Murphy"

73 Publications

High Amount of Transcription Factor IRF8 Engages AP1-IRF Composite Elements in Enhancers to Direct Type 1 Conventional Dendritic Cell Identity.

Immunity 2020 10 13;53(4):759-774.e9. Epub 2020 Aug 13.

Department of Pathology and Immunology, Washington University in St. Louis, School of Medicine, St. Louis, MO 63110, USA; Howard Hughes Medical Institute, Washington University in St. Louis, School of Medicine, St. Louis, MO 63110, USA. Electronic address:

Development and function of conventional dendritic cell (cDC) subsets, cDC1 and cDC2, depend on transcription factors (TFs) IRF8 and IRF4, respectively. Since IRF8 and IRF4 can each interact with TF BATF3 at AP1-IRF composite elements (AICEs) and with TF PU.1 at Ets-IRF composite elements (EICEs), it is unclear how these factors exert divergent actions. Here, we determined the basis for distinct effects of IRF8 and IRF4 in cDC development. Genes expressed commonly by cDC1 and cDC2 used EICE-dependent enhancers that were redundantly activated by low amounts of either IRF4 or IRF8. By contrast, cDC1-specific genes relied on AICE-dependent enhancers, which required high IRF concentrations, but were activated by either IRF4 or IRF8. IRF8 was specifically required only by a minority of cDC1-specific genes, such as Xcr1, which could distinguish between IRF8 and IRF4 DNA-binding domains. Thus, these results explain how BATF3-dependent Irf8 autoactivation underlies emergence of the cDC1-specific transcriptional program.
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http://dx.doi.org/10.1016/j.immuni.2020.07.018DOI Listing
October 2020

cDC1 prime and are licensed by CD4 T cells to induce anti-tumour immunity.

Nature 2020 08 12;584(7822):624-629. Epub 2020 Aug 12.

Department of Pathology and Immunology, Washington University School of Medicine, St Louis, MO, USA.

Conventional type 1 dendritic cells (cDC1) are thought to perform antigen cross-presentation, which is required to prime CD8 T cells, whereas cDC2 are specialized for priming CD4 T cells. CD4 T cells are also considered to help CD8 T cell responses through a variety of mechanisms, including a process whereby CD4 T cells 'license' cDC1 for CD8 T cell priming. However, this model has not been directly tested in vivo or in the setting of help-dependent tumour rejection. Here we generated an Xcr1 mouse strain to evaluate the cellular interactions that mediate tumour rejection in a model requiring CD4 and CD8 T cells. As expected, tumour rejection required cDC1 and CD8 T cell priming required the expression of major histocompatibility class I molecules by cDC1. Unexpectedly, early priming of CD4 T cells against tumour-derived antigens also required cDC1, and this was not simply because they transport antigens to lymph nodes for processing by cDC2, as selective deletion of major histocompatibility class II molecules in cDC1 also prevented early CD4 T cell priming. Furthermore, deletion of either major histocompatibility class II or CD40 in cDC1 impaired tumour rejection, consistent with a role for cognate CD4 T cell interactions and CD40 signalling in cDC1 licensing. Finally, CD40 signalling in cDC1 was critical not only for CD8 T cell priming, but also for initial CD4 T cell activation. Thus, in the setting of tumour-derived antigens, cDC1 function as an autonomous platform capable of antigen processing and priming for both CD4 and CD8 T cells and of the direct orchestration of their cross-talk that is required for optimal anti-tumour immunity.
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http://dx.doi.org/10.1038/s41586-020-2611-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7469755PMC
August 2020

An Important Role for CD4 T Cells in Adaptive Immunity to Toxoplasma gondii in Mice Lacking the Transcription Factor Batf3.

mSphere 2020 07 15;5(4). Epub 2020 Jul 15.

Department of Molecular Microbiology, Washington University in St. Louis School of Medicine, St. Louis, Missouri, USA

Immunity to at early stages of infection in C57BL/6 mice depends on gamma interferon (IFN-γ) production by NK cells, while at later stages it is primarily mediated by CD8 T cells. We decided to explore the requirement for CD4 T cells during infection in mice, which lack CD8α dendritic cells (DCs) that are necessary for cross-presentation of cell-associated antigens to CD8 T cells. We show that in this immunodeficient background on a BALB/c background, CD4 T cells become important effector cells and are able to protect mice from infection with the avirulent strain RHΔΔ Independently of the initial NK cell activation, CD4 T cells in wild-type and mice were the major source of IFN-γ. Importantly, memory CD4 T cells were sufficient to provide protective immunity following transfer into mice and secondary challenge with the virulent RHΔ strain. Collectively, these results show that under situations where CD8 cell responses are impaired, CD4 T cells provide an important alternative immune response to is a widespread parasite of animals that causes zoonotic infections in humans. Although healthy individuals generally control the infection with only moderate symptoms, it causes serious illness in newborns and those with compromised immune systems such as HIV-infected AIDS patients. Because rodents are natural hosts for , laboratory mice provide an excellent model for studying immune responses. Here, we used a combination of an attenuated mutant strain of the parasite that effectively vaccinates mice, with a defect in a transcriptional factor that impairs a critical subset of dendritic cells, to studying the immune response to infection. The findings reveal that in BALB/c mice, CD4 memory T cells play a dominant role in producing IFN-γ needed to control chronic infection. Hence, BALB/c mice may provide a more appropriate model for declining immunity seen in HIV-AIDS patients where loss of CD4 cells is associated with emergence of opportunistic infections.
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http://dx.doi.org/10.1128/mSphere.00634-20DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7364223PMC
July 2020

The MYCL and MXD1 transcription factors regulate the fitness of murine dendritic cells.

Proc Natl Acad Sci U S A 2020 03 18;117(9):4885-4893. Epub 2020 Feb 18.

Department of Pathology and Immunology, School of Medicine, Washington University in St. Louis, St. Louis, MO 63110;

We previously found that MYCL is required by a -dependent classical dendritic cell subset (cDC1) for optimal CD8 T cell priming, but the underlying mechanism has remained unclear. The MAX-binding proteins encompass a family of transcription factors with overlapping DNA-binding specificities, conferred by a C-terminal basic helix-loop-helix domain, which mediates heterodimerization. Thus, regulation of transcription by these factors is dependent on divergent N-terminal domains. The MYC family, including MYCL, has actions that are reciprocal to the MXD family, which is mediated through the recruitment of higher-order activator and repressor complexes, respectively. As potent proto-oncogenes, models of MYC family function have been largely derived from their activity at supraphysiological levels in tumor cell lines. MYC and MYCN have been studied extensively, but empirical analysis of MYCL function had been limited due to highly restricted, lineage-specific expression in vivo. Here we observed that is expressed in immature cDC1s but repressed on maturation, concomitant with induction in mature cDC1s. We hypothesized that MYCL and MXD1 regulate a shared, but reciprocal, transcriptional program during cDC1 maturation. In agreement, immature cDC1s in -deficient mice exhibited reduced expression of genes that regulate core biosynthetic processes. Mature cDC1s from mice exhibited impaired ability to inhibit the transcriptional signature otherwise supported by MYCL. The present study reveals LMYC and MXD1 as regulators of a transcriptional program that is modulated during the maturation of -dependent cDC1s.
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http://dx.doi.org/10.1073/pnas.1915060117DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7060746PMC
March 2020

Cryptic activation of an Irf8 enhancer governs cDC1 fate specification.

Nat Immunol 2019 09 12;20(9):1161-1173. Epub 2019 Aug 12.

Department of Pathology and Immunology, Washington University in St Louis, School of Medicine, St Louis, MO, USA.

Induction of the transcription factor Irf8 in the common dendritic cell progenitor (CDP) is required for classical type 1 dendritic cell (cDC1) fate specification, but the mechanisms controlling this induction are unclear. In the present study Irf8 enhancers were identified via chromatin profiling of dendritic cells and CRISPR/Cas9 genome editing was used to assess their roles in Irf8 regulation. An enhancer 32 kilobases (kb) downstream of the Irf8 transcriptional start site (+32-kb Irf8) that was active in mature cDC1s was required for the development of this lineage, but not for its specification. Instead, a +41-kb Irf8 enhancer, previously thought to be active only in plasmacytoid dendritic cells, was found to also be transiently accessible in cDC1 progenitors, and deleting this enhancer prevented the induction of Irf8 in CDPs and abolished cDC1 specification. Thus, cryptic activation of the +41-kb Irf8 enhancer in dendritic cell progenitors is responsible for cDC1 fate specification.
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http://dx.doi.org/10.1038/s41590-019-0450-xDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6707878PMC
September 2019

An Nfil3-Zeb2-Id2 pathway imposes Irf8 enhancer switching during cDC1 development.

Nat Immunol 2019 09 12;20(9):1174-1185. Epub 2019 Aug 12.

Department of Pathology and Immunology, Washington University in St. Louis, School of Medicine, St. Louis, MO, USA.

Classical type 1 dendritic cells (cDC1s) are required for antiviral and antitumor immunity, which necessitates an understanding of their development. Development of the cDC1 progenitor requires an E-protein-dependent enhancer located 41 kilobases downstream of the transcription start site of the transcription factor Irf8 (+41-kb Irf8 enhancer), but its maturation instead requires the Batf3-dependent +32-kb Irf8 enhancer. To understand this switch, we performed single-cell RNA sequencing of the common dendritic cell progenitor (CDP) and identified a cluster of cells that expressed transcription factors that influence cDC1 development, such as Nfil3, Id2 and Zeb2. Genetic epistasis among these factors revealed that Nfil3 expression is required for the transition from Zeb2 and Id2 CDPs to Zeb2 and Id2 CDPs, which represent the earliest committed cDC1 progenitors. This genetic circuit blocks E-protein activity to exclude plasmacytoid dendritic cell potential and explains the switch in Irf8 enhancer usage during cDC1 development.
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http://dx.doi.org/10.1038/s41590-019-0449-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6707889PMC
September 2019

-Dependent Genes Control Tumor Rejection Induced by Dendritic Cells Independently of Cross-Presentation.

Cancer Immunol Res 2019 01 27;7(1):29-39. Epub 2018 Nov 27.

Department of Pathology and Immunology, Washington University in St. Louis, School of Medicine, St. Louis, Missouri.

The BATF3-dependent cDC1 lineage of conventional dendritic cells (cDC) is required for rejection of immunogenic sarcomas and for rejection of progressive sarcomas during checkpoint blockade therapy. One unique function of the cDC1 lineage is the efficient cross-presentation of tumor-derived neoantigens to CD8 T cells, but it is not clear that this is the only unique function of cDC1 required for tumor rejection. We previously showed that BATF3 functions during cDC1 lineage commitment to maintain IRF8 expression in the specified cDC1 progenitor. However, since cDC1 progenitors do not develop into mature cDC1s in mice, it is still unclear whether BATF3 has additional functions in mature cDC1 cells. A transgenic -Venus reporter allele increases IRF8 protein concentration sufficiently to allow autonomous cDC1 development in spleens of mice. These restored cDC1s are transcriptionally similar to control wild-type cDC1s but have reduced expression of a restricted set of cDC1-specific genes. Restored cDC1s are able to cross-present cell-associated antigens both and However, cDC1 exhibit altered characteristics and are unable to mediate tumor rejection. These results show that BATF3, in addition to regulating expression to stabilize cDC1 lineage commitment, also controls expression of a small set of genes required for cDC1-mediated tumor rejection. These BATF3-regulated genes may be useful targets in immunotherapies aimed at promoting tumor rejection.
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http://dx.doi.org/10.1158/2326-6066.CIR-18-0138DOI Listing
January 2019

WDFY4 is required for cross-presentation in response to viral and tumor antigens.

Science 2018 11;362(6415):694-699

Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA.

During the process of cross-presentation, viral or tumor-derived antigens are presented to CD8 T cells by dependent CD8α/XCR1 classical dendritic cells (cDC1s). We designed a functional CRISPR screen for previously unknown regulators of cross-presentation, and identified the BEACH domain-containing protein WDFY4 as essential for cross-presentation of cell-associated antigens by cDC1s in mice. However, WDFY4 was not required for major histocompatibility complex class II presentation, nor for cross-presentation by monocyte-derived dendritic cells. In contrast to mice, mice displayed normal lymphoid and nonlymphoid cDC1 populations that produce interleukin-12 and protect against infection. However, similar to mice, mice failed to prime virus-specific CD8 T cells in vivo or induce tumor rejection, revealing a critical role for cross-presentation in antiviral and antitumor immunity.
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http://dx.doi.org/10.1126/science.aat5030DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6655551PMC
November 2018

Notch2-dependent DC2s mediate splenic germinal center responses.

Proc Natl Acad Sci U S A 2018 10 2;115(42):10726-10731. Epub 2018 Oct 2.

Department of Pathology and Immunology, Washington University School of Medicine in St. Louis, St. Louis, MO 63110;

CD4 T follicular helper (T) cells support germinal center (GC) reactions promoting humoral immunity. Dendritic cell (DC) diversification into genetically distinct subsets allows for specialization in promoting responses against several types of pathogens. Whether any classical DC (cDC) subset is required for humoral immunity is unknown, however. We tested several genetic models that selectively ablate distinct DC subsets in mice for their impact on splenic GC reactions. We identified a requirement for -dependent cDC2s, but not -dependent cDC1s or -dependent cDC2s, in promoting T and GC B cell formation in response to sheep red blood cells and inactivated This effect was mediated independent of and several -dependent genes expressed in cDC2s, including and Notch2 signaling during cDC2 development also substantially reduced the efficiency of cDC2s for presentation of MHC class II-restricted antigens, limiting the strength of CD4 T cell activation. Together, these results demonstrate a nonredundant role for the -dependent cDC2 subset in supporting humoral immune responses.
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http://dx.doi.org/10.1073/pnas.1809925115DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6196531PMC
October 2018

Altered compensatory cytokine signaling underlies the discrepancy between and mice.

J Exp Med 2018 05 23;215(5):1417-1435. Epub 2018 Mar 23.

Department of Pathology and Immunology, Washington University in St. Louis, School of Medicine, St. Louis, MO

The receptor Flt3 and its ligand Flt3L are both critical for dendritic cell (DC) development, but DC deficiency is more severe in mice than in mice. This has led to speculation that Flt3L binds to another receptor that also supports DC development. However, we found that Flt3L administration does not generate DCs in mice, arguing against a second receptor. Instead, DC progenitors matured in response to macrophage colony-stimulating factor (M-CSF) or stem cell factor, and deletion of in mice further reduced DC development, indicating that these cytokines could compensate for Flt3. Surprisingly, DC progenitors displayed enhanced M-CSF signaling, suggesting that loss of Flt3 increased responsiveness to other cytokines. In agreement, deletion of Flt3 in mice paradoxically rescued their severe DC deficiency. Thus, multiple cytokines can support DC development, and the discrepancy between and mice results from the increased sensitivity of progenitors to these cytokines.
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http://dx.doi.org/10.1084/jem.20171784DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5940266PMC
May 2018

Revisiting the specificity of the MHC class II transactivator CIITA in classical murine dendritic cells in vivo.

Eur J Immunol 2017 08 14;47(8):1317-1323. Epub 2017 Jul 14.

Department of Pathology and Immunology, School of Medicine, Washington University in St. Louis, St. Louis, MO, USA.

Ciita was discovered for its role in regulating transcription of major histocompatibility complex class II (MHCII) genes. Subsequently, CIITA was predicted to control many other genes based on reporter and ChIP-seq analysis but few such predictions have been verified in vivo using Ciita mice. Testing these predictions for classical dendritic cells (cDCs) has been particularly difficult, since Ciita mice lack MHCII expression required to identify cDCs. However, recent identification of the cDC-specific transcription factor Zbtb46 allows the identification of cDCs independently of MHCII expression. We crossed Zbtb46 mice onto the Ciita background and found that all cDC lineages developed in vivo in the absence of Ciita. We then compared the complete transcriptional profile of wild-type and Ciita cDCs to define the physiological footprint of CIITA for both immature and activated cDCs. We find that CIITA exerts a highly restricted control over only the MHCII, H2-DO and H2-DM genes, in DC1 and DC2 cDC subsets, but not over other proposed targets, including Ii. These findings emphasize the caveats needed in interpreting transcription factor binding sites identified by in-vitro reporter analysis, or by ChIP-seq, which may not necessarily indicate their functional activity in vivo.
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http://dx.doi.org/10.1002/eji.201747050DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5757839PMC
August 2017

Deficiency of transcription factor RelB perturbs myeloid and DC development by hematopoietic-extrinsic mechanisms.

Proc Natl Acad Sci U S A 2017 04 27;114(15):3957-3962. Epub 2017 Mar 27.

Department of Pathology and Immunology, School of Medicine, Washington University in St. Louis, St. Louis, MO 63110;

RelB is an NF-κB family transcription factor activated in the noncanonical pathway downstream of NF-κB-inducing kinase (NIK) and TNF receptor family members including lymphotoxin-β receptor (LTβR) and CD40. Early analysis suggested that RelB is required for classical dendritic cell (cDC) development based on a severe reduction of cDCs in mice associated with profound myeloid expansion and perturbations in B and T cells. Subsequent analysis of radiation chimeras generated from wild-type and bone marrow showed that RelB exerts cell-extrinsic actions on some lineages, but it has remained unclear whether the impact of RelB on cDC development is cell-intrinsic or -extrinsic. Here, we reevaluated the role of RelB in cDC and myeloid development using a series of radiation chimeras. We found that there was no cell-intrinsic requirement for RelB for development of most cDC subsets, except for the Notch2- and LTβR-dependent subset of splenic CD4 cDC2s. These results identify a relatively restricted role of RelB in DC development. Moreover, the myeloid expansion in mice resulted from hematopoietic-extrinsic actions of RelB. This result suggests that there is an unrecognized but critical role for RelB within the nonhematopoietic niche that controls normal myelopoiesis.
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http://dx.doi.org/10.1073/pnas.1619863114DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5393217PMC
April 2017

Quality of TCR signaling determined by differential affinities of enhancers for the composite BATF-IRF4 transcription factor complex.

Nat Immunol 2017 05 27;18(5):563-572. Epub 2017 Mar 27.

Department of Pathology and Immunology, Washington University in St. Louis, School of Medicine, St. Louis, Missouri, USA.

Variable strengths of signaling via the T cell antigen receptor (TCR) can produce divergent outcomes, but the mechanism of this remains obscure. The abundance of the transcription factor IRF4 increases with TCR signal strength, but how this would induce distinct types of responses is unclear. We compared the expression of genes in the T2 subset of helper T cells to enhancer occupancy by the BATF-IRF4 transcription factor complex at varying strengths of TCR stimulation. Genes dependent on BATF-IRF4 clustered into groups with distinct TCR sensitivities. Enhancers exhibited a spectrum of occupancy by the BATF-IRF4 ternary complex that correlated with the sensitivity of gene expression to TCR signal strength. DNA sequences immediately flanking the previously defined AICE motif controlled the affinity of BATF-IRF4 for direct binding to DNA. Analysis by the chromatin immunoprecipitation-exonuclease (ChIP-exo) method allowed the identification of a previously unknown high-affinity AICE2 motif at a human single-nucleotide polymorphism (SNP) of the gene encoding the immunomodulatory receptor CTLA-4 that was associated with resistance to autoimmunity. Thus, the affinity of different enhancers for the BATF-IRF4 complex might underlie divergent signaling outcomes in response to various strengths of TCR signaling.
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http://dx.doi.org/10.1038/ni.3714DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5401770PMC
May 2017

Transcription factor Zeb2 regulates commitment to plasmacytoid dendritic cell and monocyte fate.

Proc Natl Acad Sci U S A 2016 12 5;113(51):14775-14780. Epub 2016 Dec 5.

Department of Pathology and Immunology, School of Medicine, Washington University, St. Louis, MO 63110;

Dendritic cells (DCs) and monocytes develop from a series of bone-marrow-resident progenitors in which lineage potential is regulated by distinct transcription factors. Zeb2 is an E-box-binding protein associated with epithelial-mesenchymal transition and is widely expressed among hematopoietic lineages. Previously, we observed that Zeb2 expression is differentially regulated in progenitors committed to classical DC (cDC) subsets in vivo. Using systems for inducible gene deletion, we uncover a requirement for Zeb2 in the development of Ly-6C monocytes but not neutrophils, and we show a corresponding requirement for Zeb2 in expression of the M-CSF receptor in the bone marrow. In addition, we confirm a requirement for Zeb2 in development of plasmacytoid DCs but find that Zeb2 is not required for cDC2 development. Instead, Zeb2 may act to repress cDC1 progenitor specification in the context of inflammatory signals.
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http://dx.doi.org/10.1073/pnas.1611408114DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5187668PMC
December 2016

RAB43 facilitates cross-presentation of cell-associated antigens by CD8α+ dendritic cells.

J Exp Med 2016 12 29;213(13):2871-2883. Epub 2016 Nov 29.

Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110

In this study, to examine cross-presentation by classical dendritic cells (DCs; cDCs), we evaluated the role of RAB43, a protein found to be selectively expressed by Batf3-dependent CD8α and CD103 compared with other DC subsets and immune lineages. Using a specific monoclonal antibody, we localized RAB43 expression to the Golgi apparatus and LAMP1 cytoplasmic vesicles. Mice with germline or conditional deletion of Rab43 are viable and fertile and have normal development of cDCs but show a defect for in vivo and in vitro cross-presentation of cell-associated antigen. This defect is specific to cDCs, as Rab43-deficient monocyte-derived DCs showed no defect in cross-presentation of cell-associated antigen. These results suggest that RAB43 provides a specialized activity used in cross-presentation selectively by CD8α DCs but not other antigen-presenting cells.
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http://dx.doi.org/10.1084/jem.20160597DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5154939PMC
December 2016

Mafb lineage tracing to distinguish macrophages from other immune lineages reveals dual identity of Langerhans cells.

J Exp Med 2016 11 17;213(12):2553-2565. Epub 2016 Oct 17.

Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110

Current systems for conditional gene deletion within mouse macrophage lineages are limited by ectopic activity or low efficiency. In this study, we generated a Mafb-driven Cre strain to determine whether any dendritic cells (DCs) identified by Zbtb46-GFP expression originate from a Mafb-expressing population. Lineage tracing distinguished macrophages from classical DCs, neutrophils, and B cells in all organs examined. At steady state, Langerhans cells (LCs) were lineage traced but also expressed Zbtb46-GFP, a phenotype not observed in any other population. After exposure to house dust mite antigen, Zbtb46-negative CD64 inflammatory cells infiltrating the lung were substantially lineage traced, but Zbtb46-positive CD64 cells were not. These results provide new evidence for the unique identity of LCs and challenge the notion that some inflammatory cells are a population of monocyte-derived DCs.
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http://dx.doi.org/10.1084/jem.20160600DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5110021PMC
November 2016

Distinct Transcriptional Programs Control Cross-Priming in Classical and Monocyte-Derived Dendritic Cells.

Cell Rep 2016 06 2;15(11):2462-74. Epub 2016 Jun 2.

Department of Pathology and Immunology, School of Medicine, Washington University in St. Louis, St. Louis, MO 63110, USA; Howard Hughes Medical Institute, School of Medicine, Washington University in St. Louis, St. Louis, MO 63110, USA. Electronic address:

Both classical DCs (cDCs) and monocyte-derived DCs (Mo-DCs) are capable of cross-priming CD8(+) T cells in response to cell-associated antigens. We found that Ly-6C(hi)TREML4(-) monocytes can differentiate into Zbtb46(+) Mo-DCs in response to granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4) but that Ly-6C(hi)TREML4(+) monocytes were committed to differentiate into Ly-6C(lo)TREML4(+) monocytes. Differentiation of Zbtb46(+) Mo-DCs capable of efficient cross-priming required both GM-CSF and IL-4 and was accompanied by the induction of Batf3 and Irf4. However, monocytes require IRF4, but not BATF3, to differentiate into Zbtb46(+) Mo-DCs capable of cross-priming CD8(+) T cells. Instead, Irf4(-/-) monocytes differentiate into macrophages in response to GM-CSF and IL-4. Thus, cDCs and Mo-DCs require distinct transcriptional programs of differentiation in acquiring the capacity to prime CD8(+) T cells. These differences may be of consideration in the use of therapeutic DC vaccines based on Mo-DCs.
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http://dx.doi.org/10.1016/j.celrep.2016.05.025DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4941620PMC
June 2016

Transcriptional Control of Dendritic Cell Development.

Annu Rev Immunol 2016 05 23;34:93-119. Epub 2015 Dec 23.

Department of Pathology and Immunology, Washington University School of Medicine in St. Louis, Missouri 63110; email:

The dendritic cells (DCs) of the immune system function in innate and adaptive responses by directing activity of various effector cells rather than serving as effectors themselves. DCs and closely related myeloid lineages share expression of many surface receptors, presenting a challenge in distinguishing their unique in vivo functions. Recent work has taken advantage of unique transcriptional programs to identify and manipulate murine DCs in vivo. This work has assigned several nonredundant in vivo functions to distinct DC lineages, consisting of plasmacytoid DCs and several subsets of classical DCs that promote different immune effector modules in response to pathogens. In parallel, a correspondence between human and murine DC subsets has emerged, underlying structural similarities for the DC lineages between these species. Recent work has begun to unravel the transcriptional circuitry that controls the development and diversification of DCs from common progenitors in the bone marrow.
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http://dx.doi.org/10.1146/annurev-immunol-032713-120204DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5135011PMC
May 2016

Batf3 maintains autoactivation of Irf8 for commitment of a CD8α(+) conventional DC clonogenic progenitor.

Nat Immunol 2015 Jul 8;16(7):708-17. Epub 2015 Jun 8.

1] Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, Missouri, USA. [2] Howard Hughes Medical Institute, Washington University School of Medicine, St. Louis, Missouri, USA.

The transcription factors Batf3 and IRF8 are required for the development of CD8α(+) conventional dendritic cells (cDCs), but the basis for their actions has remained unclear. Here we identified two progenitor cells positive for the transcription factor Zbtb46 that separately generated CD8α(+) cDCs and CD4(+) cDCs and arose directly from the common DC progenitor (CDP). Irf8 expression in CDPs required prior autoactivation of Irf8 that was dependent on the transcription factor PU.1. Specification of the clonogenic progenitor of CD8α(+) cDCs (the pre-CD8 DC) required IRF8 but not Batf3. However, after specification of pre-CD8 DCs, autoactivation of Irf8 became Batf3 dependent at a CD8α(+) cDC-specific enhancer with multiple transcription factor AP1-IRF composite elements (AICEs) within the Irf8 superenhancer. CDPs from Batf3(-/-) mice that were specified toward development into pre-CD8 DCs failed to complete their development into CD8α(+) cDCs due to decay of Irf8 autoactivation and diverted to the CD4(+) cDC lineage.
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http://dx.doi.org/10.1038/ni.3197DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4507574PMC
July 2015

Klf4 expression in conventional dendritic cells is required for T helper 2 cell responses.

Immunity 2015 May;42(5):916-28

Department of Pathology and Immunology, Washington University School of Medicine, 660 South Euclid Avenue, St. Louis, MO 63110, USA; Howard Hughes Medical Institute, Washington University School of Medicine, St. Louis, MO 63110, USA. Electronic address:

The two major lineages of classical dendritic cells (cDCs) express and require either IRF8 or IRF4 transcription factors for their development and function. IRF8-dependent cDCs promote anti-viral and T-helper 1 (Th1) cell responses, whereas IRF4-expressing cDCs have been implicated in controlling both Th2 and Th17 cell responses. Here, we have provided evidence that Kruppel-like factor 4 (Klf4) is required in IRF4-expressing cDCs to promote Th2, but not Th17, cell responses in vivo. Conditional Klf4 deletion within cDCs impaired Th2 cell responses during Schistosoma mansoni infection, Schistosoma egg antigen (SEA) immunization, and house dust mite (HDM) challenge without affecting cytotoxic T lymphocyte (CTL), Th1 cell, or Th17 cell responses to herpes simplex virus, Toxoplasma gondii, and Citrobacter rodentium infections. Further, Klf4 deletion reduced IRF4 expression in pre-cDCs and resulted in selective loss of IRF4-expressing cDCs subsets in several tissues. These results indicate that Klf4 guides a transcriptional program promoting IRF4-expressing cDCs heterogeneity.
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http://dx.doi.org/10.1016/j.immuni.2015.04.017DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4447135PMC
May 2015

Complementary diversification of dendritic cells and innate lymphoid cells.

Curr Opin Immunol 2014 Aug 27;29:69-78. Epub 2014 May 27.

Department of Pathology and Immunology, Washington University in St. Louis, School of Medicine, 660 S. Euclid Avenue, St. Louis, MO 63110, USA; Howard Hughes Medical Institute, Washington University in St. Louis, School of Medicine, 660 S. Euclid Avenue, St. Louis, MO 63110, USA. Electronic address:

Dendritic cells (DCs) are professional antigen presenting cells conventionally thought to mediate cellular adaptive immune responses. Recent studies have led to the recognition of a non-redundant role for DCs in orchestrating innate immune responses, and in particular, for DC subset-specific interactions with innate lymphoid cells (ILCs). Recently recognized as important effectors of early immune responses, ILCs develop into subsets which mirror the transcriptional and cytokine profile of their T cell subset counterparts. DC diversification into functional subsets provides for modules of pathogen sensing and cytokine production that direct pathogen-appropriate ILC and T cell responses. This review focuses on the recent advances in the understanding of DC development, and their function in orchestrating the innate immune modules.
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http://dx.doi.org/10.1016/j.coi.2014.04.006DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5161034PMC
August 2014

Bhlhe40 controls cytokine production by T cells and is essential for pathogenicity in autoimmune neuroinflammation.

Nat Commun 2014 Apr 3;5:3551. Epub 2014 Apr 3.

Department of Pathology and Immunology, Washington University School of Medicine, St Louis, Missouri 63110, USA.

TH1 and TH17 cells mediate neuroinflammation in experimental autoimmune encephalomyelitis (EAE), a mouse model of multiple sclerosis. Pathogenic TH cells in EAE must produce the pro-inflammatory cytokine granulocyte-macrophage colony stimulating factor (GM-CSF). TH cell pathogenicity in EAE is also regulated by cell-intrinsic production of the immunosuppressive cytokine interleukin 10 (IL-10). Here we demonstrate that mice deficient for the basic helix-loop-helix (bHLH) transcription factor Bhlhe40 (Bhlhe40(-/-)) are resistant to the induction of EAE. Bhlhe40 is required in vivo in a T cell-intrinsic manner, where it positively regulates the production of GM-CSF and negatively regulates the production of IL-10. In vitro, GM-CSF secretion is selectively abrogated in polarized Bhlhe40(-/-) TH1 and TH17 cells, and these cells show increased production of IL-10. Blockade of IL-10 receptor in Bhlhe40(-/-) mice renders them susceptible to EAE. These findings identify Bhlhe40 as a critical regulator of autoreactive T-cell pathogenicity.
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http://dx.doi.org/10.1038/ncomms4551DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4016562PMC
April 2014

Heme-mediated SPI-C induction promotes monocyte differentiation into iron-recycling macrophages.

Cell 2014 Mar;156(6):1223-1234

Department of Pathology and Immunology, Washington University in St. Louis, School of Medicine, St. Louis, MO 63110, USA; Howard Hughes Medical Institute, Washington University in St. Louis, School of Medicine, St. Louis, MO 63110, USA. Electronic address:

Splenic red pulp macrophages (RPM) degrade senescent erythrocytes and recycle heme-associated iron. The transcription factor SPI-C is selectively expressed by RPM and is required for their development, but the physiologic stimulus inducing Spic is unknown. Here, we report that Spic also regulated the development of F4/80(+)VCAM1(+) bone marrow macrophages (BMM) and that Spic expression in BMM and RPM development was induced by heme, a metabolite of erythrocyte degradation. Pathologic hemolysis induced loss of RPM and BMM due to excess heme but induced Spic in monocytes to generate new RPM and BMM. Spic expression in monocytes was constitutively inhibited by the transcriptional repressor BACH1. Heme induced proteasome-dependent BACH1 degradation and rapid Spic derepression. Furthermore, cysteine-proline dipeptide motifs in BACH1 that mediate heme-dependent degradation were necessary for Spic induction by heme. These findings are the first example of metabolite-driven differentiation of a tissue-resident macrophage subset and provide new insights into iron homeostasis.
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http://dx.doi.org/10.1016/j.cell.2014.01.069DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4010949PMC
March 2014

L-Myc expression by dendritic cells is required for optimal T-cell priming.

Nature 2014 Mar 9;507(7491):243-7. Epub 2014 Feb 9.

1] Department of Pathology and Immunology, Washington University School of Medicine, 660 S. Euclid Avenue, St Louis, Missouri 63110, USA [2] Howard Hughes Medical Institute, Washington University School of Medicine, 660 S. Euclid Avenue, St Louis, Missouri 63110, USA.

The transcription factors c-Myc and N-Myc--encoded by Myc and Mycn, respectively--regulate cellular growth and are required for embryonic development. A third paralogue, Mycl1, is dispensable for normal embryonic development but its biological function has remained unclear. To examine the in vivo function of Mycl1 in mice, we generated an inactivating Mycl1(gfp) allele that also reports Mycl1 expression. We find that Mycl1 is selectively expressed in dendritic cells (DCs) of the immune system and controlled by IRF8, and that during DC development, Mycl1 expression is initiated in the common DC progenitor concurrent with reduction in c-Myc expression. Mature DCs lack expression of c-Myc and N-Myc but maintain L-Myc expression even in the presence of inflammatory signals such as granulocyte-macrophage colony-stimulating factor. All DC subsets develop in Mycl1-deficient mice, but some subsets such as migratory CD103(+) conventional DCs in the lung and liver are greatly reduced at steady state. Importantly, loss of L-Myc by DCs causes a significant decrease in in vivo T-cell priming during infection by Listeria monocytogenes and vesicular stomatitis virus. The replacement of c-Myc by L-Myc in immature DCs may provide for Myc transcriptional activity in the setting of inflammation that is required for optimal T-cell priming.
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http://dx.doi.org/10.1038/nature12967DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3954917PMC
March 2014

Notch2-dependent classical dendritic cells orchestrate intestinal immunity to attaching-and-effacing bacterial pathogens.

Nat Immunol 2013 Sep 4;14(9):937-48. Epub 2013 Aug 4.

Department of Pathology and Immunology, School of Medicine, Washington University in St. Louis, Missouri, USA.

Defense against attaching-and-effacing bacteria requires the sequential generation of interleukin 23 (IL-23) and IL-22 to induce protective mucosal responses. Although CD4(+) and NKp46(+) innate lymphoid cells (ILCs) are the critical source of IL-22 during infection, the precise source of IL-23 is unclear. We used genetic techniques to deplete mice of specific subsets of classical dendritic cells (cDCs) and analyzed immunity to the attaching-and-effacing pathogen Citrobacter rodentium. We found that the signaling receptor Notch2 controlled the terminal stage of cDC differentiation. Notch2-dependent intestinal CD11b(+) cDCs were an obligate source of IL-23 required for survival after infection with C. rodentium, but CD103(+) cDCs dependent on the transcription factor Batf3 were not. Our results demonstrate a nonredundant function for CD11b(+) cDCs in the response to pathogens in vivo.
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http://dx.doi.org/10.1038/ni.2679DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3788683PMC
September 2013

Specificity through cooperation: BATF-IRF interactions control immune-regulatory networks.

Nat Rev Immunol 2013 Jul 21;13(7):499-509. Epub 2013 Jun 21.

Department of Pathology and Immunology, Washington University School of Medicine, 660 South Euclid Avenue, St. Louis, Missouri 63110, USA.

Basic leucine zipper transcription factor ATF-like (BATF), BATF2 and BATF3 belong to the activator protein 1 (AP-1) family of transcription factors, which regulate numerous cellular processes. Initially, BATF family members were thought to function only as inhibitors of AP-1-driven transcription, but recent studies have uncovered that these factors have unique, non-redundant and positive transcriptional activities in dendritic cells, B cells and T cells. The question of how BATF and BATF3 - which lack a transcriptional activation domain, unlike the AP-1 factors FOS and JUN - can exert unique positive transcriptional specificity has now been answered by the discovery that BATF molecules interact with members of the interferon-regulatory factor (IRF) family. The capacity of the BATF leucine zipper regions to mediate dimerization with AP-1 factors and also to define cooperative interactions with heterologous factors explains both the positive transcriptional activity of BATF proteins and how they activate distinct sets of target genes compared with FOS.
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http://dx.doi.org/10.1038/nri3470DOI Listing
July 2013

Bcl11a controls Flt3 expression in early hematopoietic progenitors and is required for pDC development in vivo.

PLoS One 2013 31;8(5):e64800. Epub 2013 May 31.

Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, Missouri, United States of America.

Bcl11a is a transcription factor known to regulate lymphoid and erythroid development. Recent bioinformatic analysis of global gene expression patterns has suggested a role for Bcl11a in the development of dendritic cell (DC) lineages. We tested this hypothesis by analyzing the development of DC and other lineages in Bcl11a (-/-) mice. We found that Bcl11a was required for expression of IL-7 receptor (IL-7R) and Flt3 in early hematopoietic progenitor cells. In addition, we found severely decreased numbers of plasmacytoid dendritic cells (pDCs) in Bcl11a (-/-) fetal livers and in the bone marrow of Bcl11a (-/-) fetal liver chimeras. Moreover, Bcl11a (-/-) cells showed severely impaired in vitro development of Flt3L-derived pDCs and classical DCs (cDCs). In contrast, we found normal in vitro development of DCs from Bcl11a (-/-) fetal liver cells treated with GM-CSF. These results suggest that the persistent cDC development observed in Bcl11a (-/-) fetal liver chimeras reflects derivation from a Bcl11a- and Flt3-independent pathway in vivo.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0064800PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3669380PMC
January 2014

Therapeutic potential of B and T lymphocyte attenuator expressed on CD8+ T cells for contact hypersensitivity.

J Invest Dermatol 2013 Mar 29;133(3):702-711. Epub 2012 Nov 29.

Department of Allergy and Clinical Immunology, Graduate School of Medicine, Chiba University, Chiba, Japan. Electronic address:

In the past decade, mechanisms underlying allergic contact dermatitis have been intensively investigated by using contact hypersensitivity (CHS) models in mice. However, the regulatory mechanisms, which could be applicable for the treatment of allergic contact dermatitis, are still largely unknown. To determine the roles of B and T lymphocyte attenuator (BTLA), a CD28 family coinhibitory receptor, in hapten-induced CHS, BTLA-deficient (BTLA(-/-)) mice and littermate wild-type (WT) mice were subjected to DNFB-induced CHS, severe combined immunodeficient (SCID) mice were injected with CD4(+) T cells, and CD8(+) T cells from either WT mice or BTLA(-/-) mice were subjected to CHS. BTLA(-/-) mice showed enhanced DNFB-induced CHS and proliferation and IFN-γ production of CD8(+) T cells as compared with WT mice. SCID mice injected with WT CD4(+) T cells and BTLA(-/-) CD8(+) T cells exhibited more severe CHS as compared with those injected with WT CD4(+) T cells and WT CD8(+) T cells. On the other hand, SCID mice injected with BTLA(-/-) CD4(+) T cells and WT CD8(+) T cells exhibited similar CHS to those injected with WT CD4(+) T cells and WT CD8(+) T cells. Finally, to evaluate the therapeutic potential of an agonistic agent for BTLA on CHS, the effects of an agonistic anti-BTLA antibody (6A6) on CHS were examined. In vivo injection of 6A6 suppressed DNFB-induced CHS and IFN-γ production of CD8(+) T cells. Taken together, these results suggest that stimulation of BTLA with agonistic agents has therapeutic potential in CHS.
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http://dx.doi.org/10.1038/jid.2012.396DOI Listing
March 2013

Compensatory dendritic cell development mediated by BATF-IRF interactions.

Nature 2012 Oct 19;490(7421):502-7. Epub 2012 Sep 19.

Department of Pathology and Immunology, Washington University School of Medicine, 660 South Euclid Avenue, St Louis, Missouri 63110, USA.

The AP1 transcription factor Batf3 is required for homeostatic development of CD8α(+) classical dendritic cells that prime CD8 T-cell responses against intracellular pathogens. Here we identify an alternative, Batf3-independent pathway in mice for CD8α(+) dendritic cell development operating during infection with intracellular pathogens and mediated by the cytokines interleukin (IL)-12 and interferon-γ. This alternative pathway results from molecular compensation for Batf3 provided by the related AP1 factors Batf, which also functions in T and B cells, and Batf2 induced by cytokines in response to infection. Reciprocally, physiological compensation between Batf and Batf3 also occurs in T cells for expression of IL-10 and CTLA4. Compensation among BATF factors is based on the shared capacity of their leucine zipper domains to interact with non-AP1 factors such as IRF4 and IRF8 to mediate cooperative gene activation. Conceivably, manipulating this alternative pathway of dendritic cell development could be of value in augmenting immune responses to vaccines.
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http://dx.doi.org/10.1038/nature11531DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3482832PMC
October 2012

BATF-JUN is critical for IRF4-mediated transcription in T cells.

Nature 2012 Oct 19;490(7421):543-6. Epub 2012 Sep 19.

Laboratory of Molecular Immunology and Immunology Center, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892-1674, USA.

Interferon regulatory factor 4 (IRF4) is an IRF family transcription factor with critical roles in lymphoid development and in regulating the immune response. IRF4 binds DNA weakly owing to a carboxy-terminal auto-inhibitory domain, but cooperative binding with factors such as PU.1 or SPIB in B cells increases binding affinity, allowing IRF4 to regulate genes containing ETS-IRF composite elements (EICEs; 5'-GGAAnnGAAA-3'). Here we show that in mouse CD4(+) T cells, where PU.1/SPIB expression is low, and in B cells, where PU.1 is well expressed, IRF4 unexpectedly can cooperate with activator protein-1 (AP1) complexes to bind to AP1-IRF4 composite (5'-TGAnTCA/GAAA-3') motifs that we denote as AP1-IRF composite elements (AICEs). Moreover, BATF-JUN family protein complexes cooperate with IRF4 in binding to AICEs in pre-activated CD4(+) T cells stimulated with IL-21 and in T(H)17 differentiated cells. Importantly, BATF binding was diminished in Irf4(-/-) T cells and IRF4 binding was diminished in Batf(-/-) T cells, consistent with functional cooperation between these factors. Moreover, we show that AP1 and IRF complexes cooperatively promote transcription of the Il10 gene, which is expressed in T(H)17 cells and potently regulated by IL-21. These findings reveal that IRF4 can signal via complexes containing ETS or AP1 motifs depending on the cellular context, thus indicating new approaches for modulating IRF4-dependent transcription.
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http://dx.doi.org/10.1038/nature11530DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3537508PMC
October 2012