Mannose receptor and its putative ligands in normal murine lymphoid and nonlymphoid organs: In situ expression of mannose receptor by selected macrophages, endothelial cells, perivascular microglia, and mesangial cells, but not dendritic cells - PubMed (original) (raw)

Mannose receptor and its putative ligands in normal murine lymphoid and nonlymphoid organs: In situ expression of mannose receptor by selected macrophages, endothelial cells, perivascular microglia, and mesangial cells, but not dendritic cells

S A Linehan et al. J Exp Med. 1999.

Abstract

The mannose receptor (MR) has established roles in macrophage (Mphi) phagocytosis of microorganisms and endocytic clearance of host-derived glycoproteins, and has recently been implicated in antigen capture by dendritic cells (DCs) in vitro. MR is the founder member of a family of homologous proteins, and its recognition properties differ according to its tissue of origin. Given this heterogeneity and our recent discovery of a soluble form of MR in mouse serum, we studied the sites of synthesis of MR mRNA and expression of MR protein in normal mouse tissues. We demonstrate that synthesis and expression occur at identical sites, and that mature Mphi and endothelium are heterogeneous with respect to MR expression, additionally describing MR on perivascular microglia and glomerular mesangial cells. However, MR was not detected on DCs in situ, or on marginal zone or subcapsular sinus Mphi, both of which have MR-like binding activities. We also compared expression of MR to the binding of a recombinant probe containing the cysteine-rich domain of MR. We show that MR and its putative ligand(s) are expressed at nonoverlapping sites within lymphoid organs, consistent with a transfer function for soluble MR. Therefore, in addition to endocytic and phagocytic roles, MR may play an important role in antigen recognition and transport within lymphoid organs.

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Figures

Figure 1

Figure 1

Localization of MR and Sn mRNA in lymphoid organs by ISH. (A) MR was expressed within the medulla of lymph node (arrow), but is absent from the subcapsular sinus (arrowhead). (B) Sn expression was also seen in the medulla of the lymph node, and additionally on the subcapsular sinus. (C and D) Adjacent control sections hybridized with sense orientation probes of MR and Sn, respectively, had low levels of background. (E) MR is expressed by the red pulp of spleen. (F) In spleen, Sn was abundantly expressed by the marginal zone, but was not detected in red pulp above background levels. (G and H) Adjacent control spleen sections hybridized with sense orientation MR and Sn probes, respectively, had homogeneous levels of background throughout. (I) MR was expressed by discrete cells of the thymus. (J) Background levels of hybridization of sense orientation probe were low in thymus. Bars = 100 μm, shown in A (for A–D), E (for E–H), and I (for I and J).

Figure 2

Figure 2

Localization of MR by ICC and comparison with markers of DCs. (A) In lymph node, MR was expressed by Mφ of the medulla (m) and sinus lining Mφ, and by endothelium of the marginal sinus (ms), but not by cells of the T cell area (t). (B) In the absence of MR Ab, background staining was not detected. (C) DEC-205–expressing DCs were abundant in the T cell area in lymph node. (D) In spleen, MR was detected in Mφ and venous sinus endothelial cells of the red pulp (rp), but appeared to be absent from the marginal zone (mz) and white pulp areas (wp). Central arteriole staining was variable. (E) A control spleen section had no background labeling. (F) DCs expressing CD11c were prominent at the border of the marginal zone with the white pulp. (G) In thymus, MR immunostaining was prominent in Mφ throughout the cortex (c) and corticomedullary junction (cmj). Mφ of the medulla appeared to be negative for MR or to express very low levels (m). (H) In the absence of MR Ab, background staining in thymus was absent. (I) DEC-205 was detected on cortical epithelial cells and medullary interdigitating cells. (J) In Peyer's patch, MR expression was confined to lymphatic endothelium in the interfollicular areas (i), but was absent from follicles (f). (K) In contrast, FA.11 staining was detected in Mφ and DCs of the follicles and interfollicular areas, but was absent from endothelial cells. (L) In skin, MR was expressed by dermal Mφ, but epidermal Langerhans cell staining was not detected. (M) F4/80 was expressed by both dermal Mφ and epidermal Langerhans cells (arrow) of the skin. Bars = 50 μm, shown in A (for A–F), G (for G–I), J (for J and K), and L (for L and M).

Figure 3

Figure 3

Localization of MR with respect to markers of Mφ and endothelium by double ICC staining. MR was defined by a red product and the additional marker by blue. (A) There was some colocalization of MR with Sn, in the medullary Mφ of the lymph node (m), but Sn was also detected in MR− cells of the subcapsular sinus (arrow). (B) In lymph node, there was no colocalization between MR and CR-Fc, the latter staining only subcapsular sinus Mφ and a few cells in B cell follicles (arrowhead). (C) Scattered MR+ cells also express MHCII, and are located (D) in the paracortex adjacent to the B cell follicle defined by expression of IgM. (E) The endothelial cells that express MR in lymph node do not also express CD31, showing that only lymphatic endothelium and not high endothelial venules express MR. (F) These MR+ endothelial cells (arrow) do not coexpress macrosialin, although the intimately associated sinus lining Mφ express both markers (arrowhead). (G) In spleen, Sn was highly expressed by the marginal metallophilic Mφ and by marginal zone Mφ, and at low levels by red pulp Mφ which are strongly MR+. (H) As in lymph node, MR and CR-Fc defined two distinct populations in spleen. CR-Fc was localized in the marginal metallophilic Mφ and B cell areas of the white pulp, separated from the MR+ red pulp Mφ by the unlabeled outer marginal zone. (I) The boundary of the splenic marginal zone with the red pulp was defined by the marginal zone Mφ marker, ERTR-9. MR was absent from this population of Mφ, so its expression is restricted to the red pulp. (J) Spleen red pulp Mφ were stained with both anti-MR and FA.11, whereas the MR+ venous endothelium did not react with FA.11 (arrow). (K) In thymus, the majority of MR+ cells coexpressed the Mφ marker F4/80. (L) Staining of MR and CR-Fc in thymus revealed two distinct populations. CR-Fc was bound by large undefined cells of the medulla that may be part of the thymic epithelium, whereas MR appeared restricted to Mφ and possibly endothelium. Bars = 50 μm, shown in A (for A and B), C–F, G (for G and H), I (for I–K), and L.

Figure 4

Figure 4

Expression of MR in brain. (A) MR was detected in meningeal Mφ by ISH, here shown within an infolding of the meninges into the cerebellum. (B) By ISH, MR was detected in cells adjacent to blood vessels, the perivascular microglia. (C) ICC revealed expression of MR protein in perivascular microglia. (D) Expression of F4/ 80 by perivascular microglia confirmed that these cells were Mφ. Bars = 50 μm, shown in A (for A and B) and C (for C and D).

Figure 5

Figure 5

MR is expressed by renal glomerular mesangial cells. (A) Clusters of cells in the outer cortex of the kidney were detected by ISH using MR anti-sense probe. (B) By ICC, clusters of cells expressing MR were more easily determined to be within glomeruli. (C) A control section hybridized with MR sense probe showed no specific binding. (D) Similarly, a control section treated without MR Ab had low background staining, and none associated with glomeruli. (E) A single glomerulus stained for MR is shown at higher magnification, showing expression of MR on mesangial cells. Bars = 100 μm, shown in A (for A – D), and 50 μm, shown in E.

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