Single-Cell Survey of Human Lymphatics Unveils Marked Endothelial Cell Heterogeneity and Mechanisms of Homing for Neutrophils

Takeda, Akiko; Hollmén, Maija; Đermadi, Denis; Pan, Junliang; Brulois, Kevin; Kaukonen, Riina; Lönnberg, Tapio; Boström, Pia; Koskivuo, Ilkka; Irjala, Heikki; Miyasaka, Masayuki; Salmi, Marko; Butcher, Eugene C.; Jalkanen, Sirpa

doi:10.1016/j.immuni.2019.06.027

Cited by 163 publications

(284 citation statements)

References 56 publications

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“…We also find that individual vascular networks in LNs are composed of unique regions associated with distinct DC subsets, indicating that LN vasculature may be heterogeneous based on the local interacting cell partners. 29,30 Finally, we demonstrate the ability of CytoMAP to define microenvironments in complex disease associated tissues, including solid tumors and Mycobacterium tuberculosis (Mtb) infected murine lung granulomas. [31][32][33][34] Together, these data indicate that CytoMAP is a versatile and powerful tool for in-depth exploration of cellular positioning and tissue architecture.…”

Section: Introductionmentioning

confidence: 94%

“…These data suggest that the spatial segregation of DC subsets and their association with local blood vessels encompass discrete segments of the LN vascular branches that are preferentially associated with one or another DC population, indicating potential mechanisms for vascular heterogeneity. 29,30…”

Section: Spatially Organized Myeloid Cell Associations With Ln Blood mentioning

confidence: 99%

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CytoMAP: a spatial analysis toolbox reveals features of myeloid cell organization in lymphoid tissues

Stoltzfus

Filipek

Gern

et al. 2019

Preprint

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Recently developed approaches for highly-multiplexed 2-dimensional (2D) and 3D imaging have revealed complex patterns of cellular positioning and cell-cell interactions with important roles in both cellular and tissue level physiology. However, robust and accessible tools to quantitatively study cellular patterning and tissue architecture are currently lacking. Here, we developed a spatial analysis toolbox, Histo-Cytometric Multidimensional Analysis Pipeline (CytoMAP), which incorporates neural network based data clustering, positional correlation, dimensionality reduction, and 2D/3D region reconstruction to identify localized cellular networks and reveal fundamental features of tissue organization. We apply CytoMAP to study the microanatomy of innate immune subsets in murine lymph nodes (LNs) and reveal mutually exclusive segregation of migratory dendritic cells (DCs), regionalized compartmentalization of SIRPadermal DCs, as well as preferential association of resident DCs with select LN vasculature. These studies describe DC organization in LNs, and provide a comprehensive analytics toolbox for in-depth exploration of quantitative imaging datasets.

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Section: Introductionmentioning

confidence: 94%

Section: Spatially Organized Myeloid Cell Associations With Ln Blood mentioning

confidence: 99%

CytoMAP: a spatial analysis toolbox reveals features of myeloid cell organization in lymphoid tissues

Stoltzfus

Filipek

Gern

et al. 2019

Preprint

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“…Interestingly, cells within the LEC 2 cluster shared molecular signatures with LECs positioned at the ceiling of SCS, within the cortical sinuses, and lining lymphatic vessels, including selective expression of Emcn, Ackr4, Ackr2, Klf2, Fabp4 and Cav1 (Iftakhar et al, 2016;Takeda et al, 2019;Ulvmar et al, 2014). We also identified a subtype of dendritic-cell-like cells (Aire + APC), which likely represent the Aire-expressing ILC3-like cells that were described recently (Yamano et al, 2019).…”

Section: Scrna-seq Of Ln Cells Nominates Interacting Partners Of Ln-imentioning

confidence: 66%

“…The second blood endothelial cell cluster, termed BEC, includes a heterogeneous set of non-HEV endothelial cells, including arterial, capillary, and non-HEV venular cells (Figures S5H, S5I) (Vanlandewijck et al, 2018). Additionally, we identified two distinct populations of Prox1 + lymphatic endothelial cells (LECs), with LEC 1 likely representing a mixture of LECs lining the floor of SCS and 460 medullary sinuses based on Madcam1, Msr1, Bmp2, Vcam1, and CD274 (PD-L1) expression profiles (Cohen et al, 2014;Cordeiro et al, 2016;Takeda et al, 2019), and LEC 2 defined by unique expression of multiple extracellular matrix or structural proteins, including Fbln2, Aqp1, Fbln5, Tnc and Reln (Figures S5N, S5O).…”

Section: Scrna-seq Of Ln Cells Nominates Interacting Partners Of Ln-imentioning

confidence: 86%

“…Whether and to what extent the communication between sensory neurons and LEC 2 contributes to those processes will require a better understanding of the distribution and function of LEC 2 within LNs. Using a recently-published resource describing the transcriptional diversity and identity of LECs in human LNs, we further contextualized our 790 two murine LEC subsets (Takeda et al, 2019). Within the LEC 2 cluster, we found cells with elevated expression of Emcn, Ackr2, Ackr4, Cav1, and Fabp4, among other gene sets, which suggests high similarity with LECs found in the SCS ceiling, afferent and efferent lymphatic vessels, as well as the cortical sinuses.…”

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confidence: 85%

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Sensory Neurons Innervate Peripheral Lymph Nodes and Locally Regulate Gene Expression in Postsynaptic Endothelium, Stromal Cells, and Innate Leukocytes

Huang

Ziegler

Austin

et al. 2019

Preprint

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Immune responses within barrier tissues are regulated, in part, by nociceptors, specialized peripheral sensory neurons that detect noxious stimuli. Previous work has shown that nociceptor ablation not only alters local responses to immune challenge at peripheral sites, but also within draining lymph nodes (LNs). The mechanisms and significance of nociceptor-dependent modulation of LN function are unknown. Indeed, although sympathetic innervation of LNs is well documented, it has been unclear whether the LN parenchyma itself is innervated by sensory neurons. Here, using a combination of high-resolution imaging, retrograde viral tracing, single-cell 30 transcriptomics (scRNA-seq), and optogenetics, we identified and functionally tested a sensory neuro-immune circuit that is preferentially located in the outermost cortex of skin-draining LNs. Transcriptomic profiling revealed that there are at least four discrete subsets of sensory neurons that innervate LNs with a predominance of peptidergic nociceptors, and an innervation pattern that is distinct from that in the surrounding skin. To uncover potential LN-resident communication partners for LN-innervating sensory neurons, we employed scRNA-seq to generate a draft atlas of all murine LN cells and, based on receptor-ligand expression patterns, nominated candidate target populations among stromal and immune cells. Using selective optogenetic stimulation of LN-innervating sensory axons, we directly experimentally tested our inferred connections. Acute neuronal activation triggered rapid transcriptional changes preferentially within our top-ranked putative interacting partners, principally endothelium and other nodal stroma cells, as well as several innate leukocyte populations. Thus, LNs are monitored by a unique 40 population of sensory neurons that possesses immunomodulatory potential. 840 and Harvard. J.O

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Development of a Cancer Vaccine Using In Vivo Click‐Chemistry‐Mediated Active Lymph Node Accumulation for Improved Immunotherapy

et al. 2021

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antigens together with a strong adjuvant. Despite the promise of this approach, the clinical efficacy currently remains modest. One method to elicit a more robust and durable immune response against a particular cancer is by targeting tumor antigens and adjuvants to antigen presenting macrophages and dendritic cells (DCs). [2,3] These cells normally encounter antigens at the site of peripheral vaccination, then subsequently mature, enter the lymphatic vessels and migrate to lymph nodes (LNs) to prime effector lymphocytes. [4] Over the past decades, considerable efforts have been made to generate DC-based cancer vaccines by maturing isolated DCs with adjuvants (e.g., toll-like receptor (TLR), agonists, cytokines, and molecules that facilitate DC maturation) and equipping them with tumor antigens in vitro to improve the efficiency of these cells. [5,6] However, ex vivo cell culture is expensive and a limited amount of DCs migrate to the LNs for the activation of antigen-specific T cells. Other widely investigated strategies for allophycocyanin (APC) targeting in vivo are to fuse antigens with antibodies or ligands that target APC surface markers, such as TLRs, CD205, CD207, CD11c, and major histocompatibility complex (MHC) II, or to generate nanoformulations modified with such antibodies or ligands to deliver the antigens. [2,7] Such designs often target APCs in peripheral tissues, whereas the LN APCs are a more desirable target. LNs contain a high number of resident APCs, so delivery of antigens and adjuvants to the LN is likely to increase uptake by APCs, resulting in enhanced vaccine efficiency. [8,9] In addition, resident APCs in LNs are phenotypically immature and capable of internalizing antigens and particles. [10] Thus, LN-resident APCs may be ideal targets for immunotherapeutic drugs.Nanoformulations loaded with tumor-specific antigens and adjuvants have proven highly beneficial in boosting vaccine efficiency. [11][12][13] Those nanoformulations with diameters larger than 100 nm are primarily utilized for the delivery of antigens to peripheral APCs, which subsequently migrate to the lymph nodes to activate T cells. [14] Antigen peptide degradation, rapid MHC I turnover, and the disassociation of peptide antigen from MHC molecules may all occur during translocation to Due to their ability to elicit a potent immune reaction with low systemic toxicity, cancer vaccines represent a promising strategy for treating tumors. Considerable effort has been directed toward improving the in vivo efficacy of cancer vaccines, with direct lymph node (LN) targeting being the most promising approach. Here, a click-chemistry-based active LN accumulation system (ALAS) is developed by surface modification of lymphatic endothelial cells with an azide group, which provide targets for dibenzocyclooctyne (DBCO)-modified liposomes, to improve the delivery of encapsulated antigen and adjuvant to LNs. When loading with OVA 257-264 peptide and poly(I:C), the formulation elicits an enhanced CD8 + T cell response in vivo, resulting in ...

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Single-Cell Survey of Human Lymphatics Unveils Marked Endothelial Cell Heterogeneity and Mechanisms of Homing for Neutrophils

Cited by 163 publications

References 56 publications

CytoMAP: a spatial analysis toolbox reveals features of myeloid cell organization in lymphoid tissues

CytoMAP: a spatial analysis toolbox reveals features of myeloid cell organization in lymphoid tissues

Sensory Neurons Innervate Peripheral Lymph Nodes and Locally Regulate Gene Expression in Postsynaptic Endothelium, Stromal Cells, and Innate Leukocytes

Development of a Cancer Vaccine Using In Vivo Click‐Chemistry‐Mediated Active Lymph Node Accumulation for Improved Immunotherapy

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