Splenic red pulp macrophages are intrinsically superparamagnetic and contaminate magnetic cell isolates

Franken, Lars; Klein, Marika; Spasova, Marina; Elsukova, Anna; Wiedwald, Ulf; Welz, Meike; Knolle, Percy A.; Farle, Michael; Limmer, Andreas; Kurts, Christian

doi:10.1038/srep12940

Cited by 46 publications

(46 citation statements)

References 24 publications

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“…Here, we now demonstrate that the initially identified MR-expressing cells represent F4/80 + CD11b − CD68 + red pulp macrophages (RPMs) (28) (Fig. S7 B and C), confirming previous observations by others (29,30). However, because the role of RPMs in cross-priming is unclear, the physiological relevance of MR expression in these cells in terms of cross-presentation remains elusive.…”

Section: Resultssupporting

confidence: 70%

Mannose receptor induces T-cell tolerance via inhibition of CD45 and up-regulation of CTLA-4

Schuette

Embgenbroich

Ulas

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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The mannose receptor (MR) is an endocytic receptor involved in serum homeostasis and antigen presentation. Here, we identify the MR as a direct regulator of CD8 + T-cell activity. We demonstrate that MR expression on dendritic cells (DCs) impaired T-cell cytotoxicity in vitro and in vivo. This regulatory effect of the MR was mediated by a direct interaction with CD45 on the T cell, inhibiting its phosphatase activity, which resulted in up-regulation of cytotoxic T-lymphocyte-associated Protein 4 (CTLA-4) and the induction of T-cell tolerance. Inhibition of CD45 prevented expression of B-cell lymphoma 6 (Bcl-6), a transcriptional inhibitor that directly bound the CTLA-4 promoter and regulated its activity. These data demonstrate that endocytic receptors expressed on DCs contribute to the regulation of T-cell functionality.T he mannose receptor (MR) is an endocytic receptor belonging to the C-type lectin family and is expressed by distinct populations of dendritic cells (DCs), macrophages, and endothelial cells (1). It consists of an N-terminal cysteine-rich domain (CR), a fibronectin type II (FN II) domain, eight C-type lectinlike domains (CTLDs), a transmembrane region, and a short intracellular region. The CTLDs of the MR can bind to glycoconjugates terminated in mannose, fucose, or glucose. Despite the presence of eight CTLDs, only CTLD4 is responsible for carbohydrate binding. Additionally, the MR can bind to sulfated carbohydrates via its CR and to collagen via its FN II domain (1).In a previous study, we demonstrated that antigens internalized by the MR are processed specifically for cross-presentation (2). The correlation between the MR and cross-presentation offered the possibility that antigens targeted toward the MR could induce potent cytotoxic T-cell responses. Because the induction of a strong cytotoxic T-cell response against tumor-specific antigens is a crucial process in many tumor vaccination strategies, antigen targeting toward the MR seemed to be a promising approach. However, in in vivo tumor models, antigen targeting toward the MR did not result in a strong cytotoxic T-cell response but, instead, led to the induction of antigen-specific T-cell tolerance (3, 4). Therefore, we analyzed whether the MR has a regulatory effect on the induction of immune responses by means distinct from antigen uptake and presentation, and we investigated a direct influence of the MR on T-cell activation. We could demonstrate that the presence of the MR on the antigen-presenting cell (APC) directly impairs the cytotoxic activity of T cells in vitro and in vivo. We showed that this effect was due to a direct interaction of the MR with CD45 on the T-cell surface. This interaction inhibited CD45 activity and resulted in the up-regulation of the inhibitory molecule cytotoxic T-lymphocyte-associated Protein 4 (CTLA-4), which was responsible for the impaired cytotoxic activity of the T cells. ResultsThe Presence of the MR on DCs Reduces Cytotoxic Activity of CD8 + T Cells. To investigate whether the MR influences T-cell ...

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Section: Resultssupporting

confidence: 70%

Mannose receptor induces T-cell tolerance via inhibition of CD45 and up-regulation of CTLA-4

Schuette

Embgenbroich

Ulas

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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“…For example, resting splenic red pulp macrophages are much more prone to produce pro-inflammatory cytokines in response to exogenous stimuli as bone marrow macrophages (Wang et al, 2013). Although being more 'M1 polarized', RPM still express large amounts of the C-type lectin CD206, which is usually considered a marker for M2-polarized macrophages (Franken et al, 2015;Gordon, 2003). Because of the transcriptional diversity observed between the different macrophage subsets, it is of course not surprising that differences in the ability of these cells to react to exogenous inflammatory stimuli can be observed -it is clear that this is merely one aspect of the functional differences between these cells .…”

Section: Macrophages In Infections 477mentioning

confidence: 99%

“…The authors analysed the transcriptome of several tissue macrophage populations and tried to pinpoint the differences between macrophages and DC, as classical markers do not allow to reliably distinguish macrophages from DCs in most tissues . The integrin CD11c, for example, which is often employed as a DC marker by scientists working with mouse models, is also expressed by splenic red pulp macrophages, alveolar macrophages and Langerhans cells (Franken et al, 2015;Probst et al, 2005;Romani et al, 2003), while the wellknown macrophage marker F4/80, on the other hand, is also expressed at low levels by the most numerous splenic DC subset, the CD4 + CD11b + cDCs (Leenen et al, 1998;Franken et al, 2015). While CD64 (FcyRI) and MerTK were proposed as genes that represent a macrophage core signature, setting macrophages and DCs apart, it was initially hard for them to find genes that were expressed in all macrophage populations, but not in classical DCs.…”

Section: Macrophages In Infections 477mentioning

confidence: 99%

“…Immune surveillance (den Haan and Kraal, 2012) CD68, CD169 (Davies et al, 2013) Spleen Red pulp macrophages YS > HSC (Schulz et al, 2012) Erythrocyte recycling, iron metabolism, regulation of CD4 + T cell responses (Kohyama et al, 2009;Franken et al, 2015;Ganz, 2012;Kurotaki et al, 2011) F4/80 hi , CD206, VCAM-1, Spi-C (Franken et al, 2015) Urinary tract Urinary tract macrophages YS/HSC (Schiwon et al, 2014) Immune surveillance, regulation of neutrophil migration (Schiwon et al, 2014) F4/80, Ly6C (Schiwon et al, 2014) The origin is only stated if experimentally established. Many tissue macrophage populations show a heterogeneous origin.…”

Section: Unknownmentioning

confidence: 99%

“…To add a further level of complexity, macrophage functions do not necessarily have to be related to immune functions: red pulp macrophages of the spleen, for example, are responsible for the phagocytosis and degradation of aged or damaged erythrocytes and play a central role in the iron metabolism of the body (Davies et al, 2013;Kohyama et al, 2009). During this process, they accumulate large amounts of heme-degradation products and iron, which bestows upon them with superparamagnetic properties that can hamper certain cell-sorting techniques (Franken et al, 2015). Further examples of highly specialized macrophages are the microglia of the brain or the osteoclast in the bone.…”

Section: Introductionmentioning

confidence: 99%

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Macrophages: sentinels and regulators of the immune system

Franken¹,

Schiwon²,

Kurts³

2016

Cellular Microbiology

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146

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Summary The important role of macrophages in host defense against a variety of pathogens has long been recognized and has been documented and reviewed in numerous publications. Recently, it has become clear that tissue macrophages are not entirely derived from monocytes, as has been assumed for a long time, but rather show an ontogenetic dichotomy in most tissues: while part of the tissue macrophages are derived from monocytes, a major subset is prenatally seeded from the yolk sac. The latter subset shows a remarkable longevity and is maintained by self‐renewal in the adult animal. This paradigm shift poses interesting questions: are these two macrophage subsets functionally equivalent cells that are recruited into the tissue at different development stages, or are both macrophage subsets discrete cell types with distinct functions, which have to exist side by side? Is the functional specialization that can be observed in most macrophages due to their lineage or due to their anatomical niche? This review will give an overview about what we know of macrophage ontogeny and will discuss the influence of the macrophage lineage and location on their functional specialization.

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Characterization and Functional Analysis of Mouse Semi‐invariant Natural T Cells

et al. 2017

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Semi-invariant natural killer T (iNKT) cells are CD1d-restricted innate-like lymphocytes that recognize lipid agonists. Activated iNKT cells have immunoregulatory properties. Human and mouse iNKT cell functions elicited by different glycolipid agonists are highly conserved, making the mouse an excellent animal model for understanding iNKT cell biology in vivo. This unit describes basic methods for the characterization and quantification (see Basic Protocol 1) and functional analysis of mouse iNKT cells in vivo or in vitro. This unit also contains protocols that describe enrichment and purification of iNKT cells, generation of CD1d tetramer, and lipid antigen loading onto cell-bound and soluble CD1d for activation of NKT cell hybridomas. © 2017 by John Wiley & Sons, Inc.

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Splenic red pulp macrophages are intrinsically superparamagnetic and contaminate magnetic cell isolates

Cited by 46 publications

References 24 publications

Mannose receptor induces T-cell tolerance via inhibition of CD45 and up-regulation of CTLA-4

Mannose receptor induces T-cell tolerance via inhibition of CD45 and up-regulation of CTLA-4

Macrophages: sentinels and regulators of the immune system

Characterization and Functional Analysis of Mouse Semi‐invariant Natural T Cells

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