Development of Intravital Intermittent Confocal Imaging System for Studying Langerhans Cell Turnover

Vishwanath, Mridula; Nishibu, Akiko; Saeland, Sem; Ward, Brant R.; Mizumoto, Norikatsu; Ploegh, Hidde L.; Boes, Marianne; Takashima, Akira

doi:10.1038/sj.jid.5700448

Cited by 52 publications

(48 citation statements)

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“…To directly visualize dynamic LC behaviors in living animals, we recently developed an intravital confocal imaging system using I-Aβ-EGFP knock-in mice, in which the endogenous major histocompatibility complex class II I-Aβ chain is replaced by an EGFP-tagged version [18]. All EGFP + epidermal cells in the knock-in mice expressed CD11c as well as Langerin (CD207) and both of these LC markers were detected only in an EGFP + subpopulation of epidermal cells, allowing us to identify epidermal LCs in the absence of tissue fixation or staining [19,20]. In situ behaviors of EGFP + LCs were then visualized by recording three-dimensional confocal images every two minutes in the ear skin of anesthetized I-Aβ-EGFP knock-in mice.…”

Section: Introductionmentioning

confidence: 99%

Roles for IL-1 and TNFα in dynamic behavioral responses of Langerhans cells to topical hapten application

Nishibu

Ward

Boes

et al. 2007

Journal of Dermatological Science

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Background-To understand the behavioral biology of Langerhans cells (LCs), we recently recorded time-lapse images of LCs in the knock-in mice expressing the I-Aβ chain tagged with the enhanced green fluorescence protein (EGFP). EGFP + LCs showed relatively limited motility in the steady state, whereas topical application of dinitrofluorobenzene (DNFB) markedly augmented a unique movement of dendrites characterized by rhythmic extension and retraction, termed dSEARCH, and triggered amoeba-like lateral migration of cell bodies. Objective-To define underlying mechanisms by which hapten treatment alters LC behaviors.Methods-The I-Aβ-EGFP mice received subcutaneous (s.c.) injection of recombinant IL-1α or TNFα (50 ng/animal) and dynamic behaviors of EGFP + LCs were recorded by time-lapse confocal microscopy at several time points to measure their dSEARCH activities and lateral migration. In a different sset of experiments, IL-1 receptor antagonist (IL-1Ra) or soluble TNF receptor-2 (sTNFR2) (0.5 μg/animal) was s.c. injected into the ear skin 30 minutes before topical application of DNFB, and LC behaviors analyzed thirty hours later.Results-Local injection of IL-1α or TNFα induced significant, albeit modest, augmentation of both dSEARCH and lateral migration. Co-injection of TNFα and IL-1α further exacerbated motile activities in a synergistic manner by similar magnitudes observed after DNFB application. Conversely, DNFB-induced behavioral changes were inhibited completely by local injection of IL-1Ra or sTNFR2.Conclusion-IL-1 and TNFα serve as equally important mediators of hapten-induced alteration of LC behaviors. Motile activities of epidermal LCs are reprogrammed by selected cytokines known to be produced by keratinocytes under pathological conditions.

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Section: Introductionmentioning

confidence: 99%

Roles for IL-1 and TNFα in dynamic behavioral responses of Langerhans cells to topical hapten application

Nishibu

Ward

Boes

et al. 2007

Journal of Dermatological Science

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“…They are derived from bone marrow progenitors (26) and exhibit a capacity for self-renewal (11,36), as well as exhibiting prodigious longevity for a DC, with a half-life of up to 78 days documented (62) and, in one case, a donor's LCs were observed to persist in the recipient for more than 12 months after a skin graft procedure (23). Their generation (and/or survival) both in vitro and in vivo is acutely dependent on transforming growth factor ␤ (TGF-␤) (4, 25, 57)-TGF-␤ knockout mice do not possess LCs-and can be characterized by their (almost) unique expression of the lectin molecule, Langerin (CD207) (6,15,42,61), along with the coexpression of cutaneous leukocyte antigen, E-Cadherin, and class II major histocompatibility complex (MHC) molecules, as well as intracellular Birkbeck granules (reviewed in reference 35).…”

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Efficient Human Cytomegalovirus Reactivation Is Maturation Dependent in the Langerhans Dendritic Cell Lineage and Can Be Studied using a CD14⁺Experimental Latency Model

Huang

Kew

Jestice

et al. 2012

J Virol

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Studies from a number of laboratories have shown that the myeloid lineage is prominent in human cytomegalovirus (HCMV) latency, reactivation, dissemination, and pathogenesis. Existing as a latent infection in CD34؉ progenitors and circulating CD14 ؉ monocytes, reactivation is observed upon differentiation to mature macrophage or dendritic cell (DC) phenotypes. Langerhans' cells (LCs) are a subset of periphery resident DCs that represent a DC population likely to encounter HCMV early during primary infection. Furthermore, we have previously shown that CD34؉ derived LCs are a site of HCMV reactivation ex vivo. Accordingly, we have utilized healthy-donor CD34 ؉ cells to study latency and reactivation of HCMV in LCs. However, the increasing difficulty acquiring healthy-donor CD34؉ cells-particularly from seropositive donors due to the screening regimens used-led us to investigate the use of CD14 ؉ monocytes to generate LCs. We show here that CD14 ؉ monocytes cultured with transforming growth factor ␤ generate Langerin-positive DCs (MoLCs). Consistent with observations using CD34 ؉ derived LCs, only mature MoLCs were permissive for HCMV infection. The lytic infection of mature MoLCs is productive and results in a marked inhibition in the capacity of these cells to promote T cell proliferation. Pertinently, differentiation of experimentally latent monocytes to the MoLC phenotype promotes reactivation in a maturation and interleukin-6 (IL-6)-dependent manner. Intriguingly, however, IL-6-mediated effects were restricted to mature LCs, in contrast to observations with classical CD14؉ derived DCs. Consequently, elucidation of the molecular basis behind the differential response of the two DC subsets should further our understanding of the fundamental mechanisms important for reactivation. Human cytomegalovirus (HCMV) represents an opportunistic pathogen that is a major cause of disease in a number of immunocompromised patient populations (30,60). A significant contribution to morbidity in the clinical setting results from the reactivation of latent HCMV, particularly in seropositive bone marrow transplant recipients (40). Although the mechanisms that govern HCMV reactivation have not been fully elucidated, work from a number of laboratories has shown that CD34 ϩ hematopoietic cells, and the early granulocyte-macrophage progenitors derived from them, which are normally resident in the bone marrow are an important site for the carriage of HCMV latency in vivo (27,34,53) and that reactivation is intrinsically linked with the differentiation to a more mature myeloid cell phenotype (19,37,45,56,59,64).Langerhans' cells (LCs) are a unique population of dendritic cells (DCs) resident in the epidermis and a number of mucosal tissues (e.g., nasal, oral, vaginal, and corneal). They are derived from bone marrow progenitors (26) and exhibit a capacity for self-renewal (11, 36), as well as exhibiting prodigious longevity for a DC, with a half-life of up to 78 days documented (62) and, in one case, a donor's LCs were observed to persis...

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“…In agreement with a recent report, stainings with either anti-MHCII or anti-Langerin were identical (data not shown), as all MHCIIpositive cells in murine epidermis were shown to coexpress Langerin. 26 LN cell suspensions were generated by collagenase digestion as described previously 27 and immunostained using APC-conjugated anti-MHCII, PE-conjugated anti-PD-L1, clone MIH5 (both from eBioscience, San Diego, CA) and Alexa488-conjugated anti-Langerin (clone 923F3 from AbCys). …”

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Mast cells are crucial for early inflammation, migration of Langerhans cells, and CTL responses following topical application of TLR7 ligand in mice

Heib

Becker

Warger

et al. 2007

Blood

103

108

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Until recently, IgE-activated mast cells have been regarded merely as effector cells of adaptive immune responses, involved in allergic reactions and mucosal immunity to parasites. Herein, we report that murine dermal mast cells, activated by local administration of a cream containing the synthetic TLR7 ligand imiquimod, are essential to initiate an early inflammatory reaction. The mast-cell-derived cytokines TNF-␣ and IL-1␤ play an important role in this process. Furthermore, TLR7-activated mast cells are also able to promote the emigration of Langerhans cells, which partly depends on the expression of mast-cell-derived IL-1␤. We have previously shown that TLR7 ligation enhances transcutaneous immunization evoked by topical application of vaccine antigens to the skin, a procedure that directly targets skin-resident antigen-presenting cells.Consequently, we now demonstrate here that the capacity to mount a peptidespecific cytotoxic T-lymphocyte response following transcutaneous immunization using imiquimod as adjuvant is severely impaired in mast-cell-deficient mice. IntroductionIgE-dependent reactions including dysregulated allergic responses to environmental antigens and mucosal immunity to some extracellular parasites were believed to be the hallmarks of mast-cell immunology for several decades.We know by now that mast cells, predominantly localized at the interface between host and environment (ie, skin and mucosal surfaces), are in addition able to perceive a variety of allergens and invading pathogens. Based on 2 groundbreaking publications using murine models of acute bacterial infection, mast cells were found to be critical effectors of innate immunity. In these studies, mastcell-deficient mice were highly susceptible to induced septic peritonitis 1 and Klebsiella pneumoniae instillation. 2 In these settings, the unique ability of mast cells to secrete preformed TNF-␣ within minutes following IgE-independent stimulation enables the host to mount an early and protective neutrophil response to bacterial challenge. Evidence further accumulates that mast cells are implicated in host defense against a still increasing range of clinically relevant bacterial infections. [3][4][5] Furthermore, mast cells are also thought to participate in response to viruses, but this is less understood and deserves further research. 5,6 With respect to IgEindependent activation, both rodent and human mast cells were found to express a variety of Toll-like receptors, some of which are expressed only on certain mast cell subsets. [5][6][7] However, several murine in vivo models clearly showed the importance of TLRmediated mast cell activation, for example, for the clearance of enterobacterial infection by triggering TLR4 8 and for the recruitment of CD8 ϩ T cells into the peritoneal cavity following ligation of TLR3 with poly(I:C), which mimics viral dsRNA. 9 It is also increasingly being appreciated that mast cells are able to finely control the magnitude of the secretory response following activation. 10 Thus, IgE-independent ...

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Development of Intravital Intermittent Confocal Imaging System for Studying Langerhans Cell Turnover

Cited by 52 publications

References 30 publications

Roles for IL-1 and TNFα in dynamic behavioral responses of Langerhans cells to topical hapten application

Roles for IL-1 and TNFα in dynamic behavioral responses of Langerhans cells to topical hapten application

Efficient Human Cytomegalovirus Reactivation Is Maturation Dependent in the Langerhans Dendritic Cell Lineage and Can Be Studied using a CD14⁺Experimental Latency Model

Mast cells are crucial for early inflammation, migration of Langerhans cells, and CTL responses following topical application of TLR7 ligand in mice

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Development of Intravital Intermittent Confocal Imaging System for Studying Langerhans Cell Turnover

Cited by 52 publications

References 30 publications

Roles for IL-1 and TNFα in dynamic behavioral responses of Langerhans cells to topical hapten application

Roles for IL-1 and TNFα in dynamic behavioral responses of Langerhans cells to topical hapten application

Efficient Human Cytomegalovirus Reactivation Is Maturation Dependent in the Langerhans Dendritic Cell Lineage and Can Be Studied using a CD14+Experimental Latency Model

Mast cells are crucial for early inflammation, migration of Langerhans cells, and CTL responses following topical application of TLR7 ligand in mice

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Efficient Human Cytomegalovirus Reactivation Is Maturation Dependent in the Langerhans Dendritic Cell Lineage and Can Be Studied using a CD14⁺Experimental Latency Model