scholarly journals Lymphocyte recognition of high endothelium: antibodies to distinct epitopes of an 85-95-kD glycoprotein antigen differentially inhibit lymphocyte binding to lymph node, mucosal, or synovial endothelial cells.

1987 ◽  
Vol 105 (2) ◽  
pp. 983-990 ◽  
Author(s):  
S Jalkanen ◽  
R F Bargatze ◽  
J de los Toyos ◽  
E C Butcher
Keyword(s):  
Monoclonal Antibody ◽  
Lymph Node ◽  
Human Lymphocyte ◽  
Molecular Mechanisms ◽  
Polyclonal Antiserum ◽  
Surface Glycoprotein ◽  
Lymphoid Tissues ◽  
Specific Cell ◽  
High Endothelial Venules ◽  
Glycoprotein Antigen

The tissue-specific homing of lymphocytes is directed by specialized high endothelial venules (HEV). At least three functionally independent lymphocyte/HEV recognition systems exist, controlling the extravasation of circulating lymphocytes into peripheral lymph nodes, mucosal lymphoid tissues (Peyer's patches or appendix), and the synovium of inflamed joints. We report here that antibodies capable of inhibiting human lymphocyte binding to one or more HEV types recognize a common 85-95-kD lymphocyte surface glycoprotein antigen, defined by the non-blocking monoclonal antibody, Hermes-1. We demonstrate that MEL-14, a monoclonal antibody against putative lymph node "homing receptors" in the mouse, functionally inhibits human lymphocyte binding to lymph node HEV but not to mucosal or synovial HEV, and cross-reacts with the 85-95-kD Hermes-1 antigen. Furthermore, we show that Hermes-3, a novel antibody produced by immunization with Hermes-1 antigen isolated from a mucosal HEV-specific cell line, selectively blocks lymphocyte binding to mucosal HEV. Such tissue specificity of inhibition suggests that MEL-14 and Hermes-3 block the function of specific lymphocyte recognition elements for lymph node and mucosal HEV, respectively. Recognition of synovial HEV also involves the 85-95-kD Hermes-1 antigen, in that a polyclonal antiserum produced against the isolated antigen blocks all three classes of lymphocyte-HEV interaction. From these studies, it is likely that the Hermes-1-defined 85-95-kD glycoprotein class either comprises a family of related but functionally independent receptors for HEV, or associates both physically and functionally with such receptors. The findings imply that related molecular mechanisms are involved in several functionally independent cell-cell recognition events that direct lymphocyte traffic.

scholarly journals Flow cytometric analysis of the Hermes homing-associated antigen on human lymphocyte subsets

Blood ◽  
1989 ◽  
Vol 74 (2) ◽  
pp. 751-760 ◽  
Author(s):  
J de los Toyos ◽  
S Jalkanen ◽  
EC Butcher
Keyword(s):  
T Cells ◽  
T Cell ◽  
B Cells ◽  
Human Lymphocyte ◽  
High Surface ◽  
Interleukin 2 ◽  
Flow Cytometric Analysis ◽  
Surface Glycoprotein ◽  
Lymphoid Tissues ◽  
High Endothelial Venules

Abstract The homing of lymphocytes is controlled by interactions with high endothelial venules (HEV), specialized vessels that define sites of lymphocyte extravasation into lymph nodes and inflamed tissues. In humans, lymphocyte-HEV binding involves a lymphocyte surface glycoprotein (GP) of 85 to 95 kd (CD44, H-CAM), defined by monoclonal antibody (MoAb) Hermes-1. To define the expression of this homing- associated adhesion molecule during human lymphocyte development, we performed two-color immunofluorescence analyses of human bone marrow (BM), thymus, peripheral blood (PB), and tonsillar lymphocytes. The highest levels of Hermes-1 antigen are displayed by circulating B and T cells in the blood, which are uniformly positive and bear roughly twice the level of antigen present on mature lymphocytes within organized lymphoid tissues and BM. “Immature” (CD4+, CD8+) T cells in the thymus are Hermes-1lo to-, whereas thymocytes of mature phenotype (CD4+ or CD8+) are positive. The Hermes-1 antigen is present at high levels on the same population of thymocytes that bears high surface levels of CD3, a component of the T-cell antigen receptor complex, suggesting that levels of T-cell homing and antigen receptors characteristic of mature peripheral T cells appear coordinately during thymocyte maturation/selection. Essentially all T cells in the periphery are Hermes-1hi, including T blasts, and the homing-associated antigen is maintained at high levels on T cells stimulated in vitro by phytohemagglutinin (PHA) and on interleukin-2 (IL-2) maintained T-cell clones and lines. In contrast, although most resting IgD+ B cells are positive a significant fraction of B cells in tonsils are Hermes-1lo to- ; these cells are predominantly PNAhi, IgD-, and CD20hi, a phenotype characteristic of sessile, activated B cells in germinal centers. In all lymphocyte populations examined, there is a linear correlation in staining for Hermes-1 and for Hermes-3, an antibody that defines a distinct functionally important epitope on this molecule. The results demonstrate a precise regulation of this homing-associated antigen during lymphocyte differentiation.

scholarly journals Flow cytometric analysis of the Hermes homing-associated antigen on human lymphocyte subsets

Blood ◽  
1989 ◽  
Vol 74 (2) ◽  
pp. 751-760
Author(s):  
J de los Toyos ◽  
S Jalkanen ◽  
EC Butcher
Keyword(s):  
T Cells ◽  
T Cell ◽  
B Cells ◽  
Human Lymphocyte ◽  
High Surface ◽  
Interleukin 2 ◽  
Flow Cytometric Analysis ◽  
Surface Glycoprotein ◽  
Lymphoid Tissues ◽  
High Endothelial Venules

The homing of lymphocytes is controlled by interactions with high endothelial venules (HEV), specialized vessels that define sites of lymphocyte extravasation into lymph nodes and inflamed tissues. In humans, lymphocyte-HEV binding involves a lymphocyte surface glycoprotein (GP) of 85 to 95 kd (CD44, H-CAM), defined by monoclonal antibody (MoAb) Hermes-1. To define the expression of this homing- associated adhesion molecule during human lymphocyte development, we performed two-color immunofluorescence analyses of human bone marrow (BM), thymus, peripheral blood (PB), and tonsillar lymphocytes. The highest levels of Hermes-1 antigen are displayed by circulating B and T cells in the blood, which are uniformly positive and bear roughly twice the level of antigen present on mature lymphocytes within organized lymphoid tissues and BM. “Immature” (CD4+, CD8+) T cells in the thymus are Hermes-1lo to-, whereas thymocytes of mature phenotype (CD4+ or CD8+) are positive. The Hermes-1 antigen is present at high levels on the same population of thymocytes that bears high surface levels of CD3, a component of the T-cell antigen receptor complex, suggesting that levels of T-cell homing and antigen receptors characteristic of mature peripheral T cells appear coordinately during thymocyte maturation/selection. Essentially all T cells in the periphery are Hermes-1hi, including T blasts, and the homing-associated antigen is maintained at high levels on T cells stimulated in vitro by phytohemagglutinin (PHA) and on interleukin-2 (IL-2) maintained T-cell clones and lines. In contrast, although most resting IgD+ B cells are positive a significant fraction of B cells in tonsils are Hermes-1lo to- ; these cells are predominantly PNAhi, IgD-, and CD20hi, a phenotype characteristic of sessile, activated B cells in germinal centers. In all lymphocyte populations examined, there is a linear correlation in staining for Hermes-1 and for Hermes-3, an antibody that defines a distinct functionally important epitope on this molecule. The results demonstrate a precise regulation of this homing-associated antigen during lymphocyte differentiation.

scholarly journals Demonstration that a lectin-like receptor (gp90MEL) directly mediates adhesion of lymphocytes to high endothelial venules of lymph nodes.

1989 ◽  
Vol 109 (5) ◽  
pp. 2463-2469 ◽  
Author(s):  
J S Geoffroy ◽  
S D Rosen
Keyword(s):  
Lymph Node ◽  
Lymphoid Organ ◽  
Lymphoid Organs ◽  
Surface Glycoprotein ◽  
Lymphocyte Migration ◽  
Peripheral Lymph Node ◽  
Adhesive Interaction ◽  
High Endothelial Venules ◽  
Cell Surface Glycoprotein ◽  
Peripheral Lymph

Lymphocyte migration from the blood into most secondary lymphoid organs is initiated by a highly selective adhesive interaction with the endothelium of specialized blood vessels known as high endothelial venules (HEV). The propensity of lymphocytes to migrate to particular lymphoid organs is known as lymphocyte homing, and the receptors on lymphocytes that dictate interactions with HEV at particular anatomical sites are designated "homing receptors". Based upon antibody blockade experiments and cell-type distribution studies, a prominent candidate for the peripheral lymph node homing receptor in mouse is the approximately 90-kD cell surface glycoprotein (gp90MEL) recognized by the monoclonal antibody MEL-14. Previous work, including sequencing of a cDNA encoding for this molecule, supports the possibility that gp90MEL is a calcium-dependent lectin-like receptor. Here, we show that immunoaffinity-purified gp90MEL interacts in a sugar-inhibitable manner with sites on peripheral lymph node HEV and prevents attachment of lymphocytes. Lymphocyte attachment to HEV in Peyer's patches, a gut-associated lymphoid organ, is not affected by gp90MEL. The results demonstrate that gp90MEL, as a lectin-like receptor, directly bridges lymphocytes to the endothelium.

scholarly journals Anti-CD43 Inhibits Monocyte-Endothelial Adhesion in Inflammation and Atherogenesis

Blood ◽  
1997 ◽  
Vol 90 (9) ◽  
pp. 3587-3594 ◽  
Author(s):  
Leslie M. McEvoy ◽  
Mark A. Jutila ◽  
Philip S. Tsao ◽  
John P. Cooke ◽  
Eugene C. Butcher
Keyword(s):  
Lymph Node ◽  
Ex Vivo ◽  
Inhibitory Effect ◽  
Lymphoid Tissues ◽  
Monocyte Adhesion ◽  
Blood Monocytes ◽  
High Endothelial Venules ◽  
Aortic Endothelial Cells ◽  
Novel Target ◽  
And Migration

Abstract Recruitment of blood monocytes into tissues is a central event in the inflammatory response and in atherogenesis. The mechanisms leading to monocyte adhesion and migration through endothelium are not completely defined. We recently reported that MAb L11, against the leukocyte sialomucin CD43, blocks T-lymphocyte binding to lymph node and Peyer's patch high endothelial venules (HEV) and inhibits T-cell extravasation from the blood into organized secondary lymphoid tissues. We have now assessed the ability of L11 to inhibit monocyte-endothelial (EC) interactions and trafficking. L11 blocks binding of WEHI78/24 cells, a murine monocytoid cell line, to inflamed lymph node HEV and inhibits recruitment of monocytes and neutrophils to thioglycollate-inflamed peritoneum. Because monocyte adhesion to the endothelium and diapedesis in lesion-prone regions of the vasculature is among the earliest events in atherogenesis, leading to formation of lipid-laden foam cells, the ability of L11 to block monocyte recognition of aortic endothelial cells was assessed in a novel ex vivo assay of monocyte binding to intact rabbit aortic endothelium. Cholesterol feeding of rabbits induces enhanced aortic adhesiveness for monocytes and WEHI78/24 monocytoid cells, and this adhesion is inhibited by L11. The inhibitory effect of L11 is additive with that of a cocktail of anti–L-selectin and anti-α4 and β2 integrin monoclonal antibodies. Thus, CD43 represents a novel target for manipulation of monocyte recruitment in inflammation and atherogenesis.

scholarly journals Localization of ligands for L-selectin in mouse peripheral lymph node high endothelial cells by colloidal gold conjugates

Blood ◽  
1994 ◽  
Vol 84 (11) ◽  
pp. 3766-3775 ◽  
Author(s):  
A Kikuta ◽  
SD Rosen
Keyword(s):  
Endothelial Cells ◽  
Lymph Node ◽  
Colloidal Gold ◽  
Secretory Granules ◽  
Polyclonal Antiserum ◽  
Secretory Product ◽  
Electron Microscopic ◽  
High Endothelial Venules ◽  
Ultrastructural Studies ◽  
Peripheral Lymph

L-selectin, a Ca(2+)-dependent lectin-like receptor, mediates lymphocyte attachment to high endothelial venules (HEV) of peripheral lymph nodes (PLN) during the process of lymphocyte homing. Two endothelial-derived ligands for L-selectin, known as GlyCAM-1 (Sgp50) and CD34 (Sgp90), have been identified by affinity precipitation of lymph node extracts with a chimeric molecule that combines the extracellular domains of L-selectin with the human IgG1 Fc region (L- selectin-IgG) (J Cell Biol 110:2221, 1990). Here, using a histologic probe based on colloidal gold conjugated to L-selectin-IgG (LS-Ig), we performed morphologic mapping of the HEV ligands in PLN at both the light and electron microscopic levels. With a postembedding labeling method, intense LS-Ig-gold staining of PLN HEV was observed, while the HEV of Peyer's patches (PP) were negative. The specificity of LS-Ig- gold staining was established by pretreatment of sections with sialidase and coincubation of sections with EGTA, fucoidin, or L- selectin-IgG itself. In ultrastructural studies of high endothelial cells(HEC), gold particles were bound to the trans-Golgi network(TGN) and to peripheral vesicles in the cytoplasm. Gold labeling was also detected in a patchy distribution on the entire luminal vascular surface of HEC. Although the perivascular fibroreticular sheath of HEV was frequently labeled limited labeling was observed on the basolateral surfaces of the HEC. In most cases, the HEC membrane surrounding migrating lymphocytes was negative. These results show that L-selectin ligands pass through the Golgi apparatus during their biosynthesis, are stored in secretory granules, and are expressed on the vascular luminal surface of the HEC. A polyclonal antiserum to GlyCAM-1 intensely stained intracellular organelles in the biosynthetic pathway including cytoplasmic vesicles, but failed to stain the cell surface of HEC. Given its presence in serum as a soluble factor, GlyCAM-1 is likely to be a secretory product.

scholarly journals Sulfation-dependent recognition of high endothelial venules (HEV)-ligands by L-selectin and MECA 79, and adhesion-blocking monoclonal antibody.

1994 ◽  
Vol 180 (6) ◽  
pp. 2219-2226 ◽  
Author(s):  
S Hemmerich ◽  
E C Butcher ◽  
S D Rosen
Keyword(s):  
Monoclonal Antibody ◽  
Lymph Node ◽  
Lymph Nodes ◽  
Lymphoid Organs ◽  
Metabolic Inhibitor ◽  
Dependent Manner ◽  
Independent Molecule ◽  
High Endothelial Venules ◽  
Additional Ligand ◽  
Secondary Lymphoid Organs

L-selectin is a lectin-like receptor that mediates the attachment of lymphocytes to high endothelial venules (HEV) of lymph nodes during the process of lymphocyte recirculation. Two sulfated, mucin-like glycoproteins known as Sgp50/GlyCAM-1 and Sgp90/CD34 have previously been identified as HEV-associated ligands for L-selectin. These proteins were originally detected with an L-selectin/Ig chimera called LEC-IgG. GlyCAM-1 and CD34 are also recognized by an antiperipheral node addressin (PNAd) mAb called MECA 79, which blocks L-selectin-dependent adhesion and selectively stains lymph node HEV. The present study compares the requirements for the binding of MECA 79 and LEC-IgG to HEV-ligands. Whereas desialylation of GlyCAM-1 and CD34 drastically reduced binding to LEC-IgG, this treatment enhanced the binding of GlyCAM-1 to MECA 79. In contrast, the binding of both MECA 79 and LEC-IgG to GlyCAM-1 and CD34 was greatly decreased when the sulfation of these ligands was reduced with chlorate, a metabolic inhibitor of sulfation. Because MECA 79 stains HEV-like vessels at various sites of inflammation, recognition by L-selectin of ligands outside of secondary lymphoid organs may depend on sulfation. In addition to their reactivity with GlyCAM-1 and CD34, both MECA 79 and LEC-IgG recognize an independent molecule of approximately 200 kD in a sulfate-dependent manner. Thus, this molecule, which we designate Sgp200, is an additional ligand for L-selectin.

Fever-range hyperthermia dynamically regulates lymphocyte delivery to high endothelial venules

Blood ◽  
2001 ◽  
Vol 97 (9) ◽  
pp. 2727-2733 ◽  
Author(s):  
Sharon S. Evans ◽  
Wan-Chao Wang ◽  
Mark D. Bain ◽  
Randy Burd ◽  
Julie R. Ostberg ◽  
...  
Keyword(s):  
Lymph Node ◽  
Acute Infection ◽  
Fold Increase ◽  
Whole Body ◽  
Lymphoid Tissues ◽  
Hyperthermia Treatment ◽  
High Endothelial Venules ◽  
Adhesive Interactions

Abstract Fever is associated with increased survival during acute infection, although its mechanism of action is largely unknown. This study found evidence of an unexpectedly integrated mechanism by which fever-range temperatures stimulate lymphocyte homing to secondary lymphoid tissues by increasing L-selectin and α4β7 integrin–dependent adhesive interactions between circulating lymphocytes and specialized high endothelial venules (HEV). Exposure of splenic lymphocytes in vivo to fever-like whole-body hyperthermia (WBH; 39.8 ± 0.2°C for 6 hours) stimulated both L-selectin and α4β7 integrin–dependent adhesion of lymphocytes to HEV under shear conditions in lymph nodes and Peyer patches. The adhesiveness of HEV ligands for L-selectin and α4β7 integrin (ie, peripheral lymph node addressin and mucosal addressin cell adhesion molecule-1) also increased during WBH or febrile responses associated with lipopolysaccharide-induced or turpentine-induced inflammation. Similar increases in HEV adhesion occurred during hyperthermia treatment of lymph node and Peyer patch organ cultures in vitro, indicating that the local lymphoid tissue microenvironment is sufficient for the hyperthermia response. In contrast, WBH did not augment adhesion in squamous endothelium of nonlymphoid tissues. Analysis of homing of α4β7hi L-selectinlo murine TK1 cells and L-selectinhi α4β7 integrin-negative 300.19/L-selectin transfectant cells showed that fever-range temperatures caused a 3- to 4-fold increase in L-selectin and α4β7 integrin–dependent trafficking to secondary lymphoid tissues. Thus, enhanced lymphocyte delivery to HEV by febrile temperatures through bimodal regulation of lymphocyte and endothelial adhesion provides a novel mechanism to promote immune surveillance.

In vitro analysis of the homing properties of human lymphocytes: developmental regulation of functional receptors for high endothelial venules

Blood ◽  
1985 ◽  
Vol 66 (3) ◽  
pp. 577-582
Author(s):  
ST Jalkanen ◽  
EC Butcher
Keyword(s):  
Human Lymphocyte ◽  
In Vitro Model ◽  
Human Lymphocytes ◽  
Surrounding Tissue ◽  
Lymphoid Tissues ◽  
High Endothelial Venules ◽  
Circulating Lymphocytes ◽  
Vitro Model

Circulating lymphocytes leave the blood by binding to specialized high endothelial cells lining postcapillary venules in lymphoid organs or sites of chronic inflammations, migrating through the vessel wall into the surrounding tissue. The capacity of lymphocytes to recognize and bind to high endothelial venules (HEVs) is thus central to the overall process of lymphocyte traffic and recirculation. We show that viable human lymphocytes bind selectively to HEVs in frozen sections of normal human lymph nodes, thus defining a simple in vitro model for the study of human lymphocyte homing properties. Optimal conditions for the quantitative analysis of lymphocyte-HEV interaction are described. Furthermore, by using this assay, we demonstrate that the ability of human lymphocyte populations to bind to HEVs parallels their presumed migratory status in vivo. Thus, thymocytes and bone marrow cells, which are sessile in vivo, bind poorly to HEVs in comparison with mature circulating lymphocytes in peripheral blood or in peripheral lymphoid tissues. These results indicate that HEV-binding ability is a regulated property of mature lymphocytes and, as demonstrated previously in animal models, probably plays a fundamental role in controlling lymphocyte traffic in humans. The in vitro model of lymphocyte-HEV interaction thus provides a unique means to assay the migratory properties of normal and neoplastic human lymphocyte subsets, to analyze the role of lymphocyte traffic mechanisms in normal and pathologic inflammatory reactions, and to define some of the molecular mechanisms responsible for the control of lymphocyte migration and positioning in humans.

scholarly journals In vitro analysis of the homing properties of human lymphocytes: developmental regulation of functional receptors for high endothelial venules

Blood ◽  
1985 ◽  
Vol 66 (3) ◽  
pp. 577-582 ◽  
Author(s):  
ST Jalkanen ◽  
EC Butcher
Keyword(s):  
Human Lymphocyte ◽  
In Vitro Model ◽  
Human Lymphocytes ◽  
Surrounding Tissue ◽  
Lymphoid Tissues ◽  
High Endothelial Venules ◽  
Circulating Lymphocytes ◽  
Vitro Model

Abstract Circulating lymphocytes leave the blood by binding to specialized high endothelial cells lining postcapillary venules in lymphoid organs or sites of chronic inflammations, migrating through the vessel wall into the surrounding tissue. The capacity of lymphocytes to recognize and bind to high endothelial venules (HEVs) is thus central to the overall process of lymphocyte traffic and recirculation. We show that viable human lymphocytes bind selectively to HEVs in frozen sections of normal human lymph nodes, thus defining a simple in vitro model for the study of human lymphocyte homing properties. Optimal conditions for the quantitative analysis of lymphocyte-HEV interaction are described. Furthermore, by using this assay, we demonstrate that the ability of human lymphocyte populations to bind to HEVs parallels their presumed migratory status in vivo. Thus, thymocytes and bone marrow cells, which are sessile in vivo, bind poorly to HEVs in comparison with mature circulating lymphocytes in peripheral blood or in peripheral lymphoid tissues. These results indicate that HEV-binding ability is a regulated property of mature lymphocytes and, as demonstrated previously in animal models, probably plays a fundamental role in controlling lymphocyte traffic in humans. The in vitro model of lymphocyte-HEV interaction thus provides a unique means to assay the migratory properties of normal and neoplastic human lymphocyte subsets, to analyze the role of lymphocyte traffic mechanisms in normal and pathologic inflammatory reactions, and to define some of the molecular mechanisms responsible for the control of lymphocyte migration and positioning in humans.

scholarly journals The human peripheral lymph node vascular addressin is a ligand for LECAM-1, the peripheral lymph node homing receptor.

1991 ◽  
Vol 114 (2) ◽  
pp. 343-349 ◽  
Author(s):  
E L Berg ◽  
M K Robinson ◽  
R A Warnock ◽  
E C Butcher
Keyword(s):  
Lymph Node ◽  
Lymphoid Tissues ◽  
Dependent Manner ◽  
Peripheral Lymph Node ◽  
Lymphocyte Homing ◽  
High Endothelial Venules ◽  
Homing Receptor ◽  
Calcium Dependent ◽  
Lymph Node Homing ◽  
Peripheral Lymph

The trafficking of lymphocytes from the blood and into lymphoid organs is controlled by tissue-selective lymphocyte interactions with specialized endothelial cells lining post capillary venules, in particular the high endothelial venules (HEV) found in lymphoid tissues and sites of chronic inflammation. Lymphocyte interactions with HEV are mediated in part by lymphocyte homing receptors and tissue-specific HEV determinants, the vascular addressins. A peripheral lymph node addressin (PNAd) has been detected immunohistologically in mouse and man by monoclonal antibody MECA-79, which inhibits lymphocyte homing to lymph nodes and lymphocyte binding to lymph node and tonsillar HEV. The human MECA-79 antigen, PNAd, is molecularly distinct from the 65-kD mucosal vascular addressin. The most abundant iodinated species by SDS-PAGE is 105 kD. When affinity isolated and immobilized on glass slides, MECA-79 immunoisolated material binds human and mouse lymphocytes avidly in a calcium dependent manner. Binding is blocked by mAb MECA-79, by antibodies against mouse or human LECAM-1 (the peripheral lymph node homing receptor, the MEL-14 antigen, LAM-1), and by treatment of PNAd with neuraminidase. Expression of LECAM-1 cDNA confers PNAd binding ability on a transfected B cell line. We conclude that LECAM-1 mediates lymphocyte binding to PNAd, an interaction that involves the lectin activity of LECAM-1 and carbohydrate determinants on the addressin.

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