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LYVE-1, a new homologue of the CD44 glycoprotein, is a lymph-specific receptor for hyaluronan.

Banerji S, Ni J, Wang SX, Clasper S, Su J, Tammi R, Jones M, Jackson DG - J. Cell Biol. (1999)

Bottom Line: Like CD44, the LYVE-1 molecule binds both soluble and immobilized HA.However, unlike CD44, the LYVE-1 molecule colocalizes with HA on the luminal face of the lymph vessel wall and is completely absent from blood vessels.Hence, LYVE-1 is the first lymph-specific HA receptor to be characterized and is a uniquely powerful marker for lymph vessels themselves.

View Article: PubMed Central - PubMed

Affiliation: University of Oxford, Molecular Immunology Group, Nuffield Department of Medicine, John Radcliff Hospital, Headington, Oxford OX3 9DU, United Kingdom.

ABSTRACT
The extracellular matrix glycosaminoglycan hyaluronan (HA) is an abundant component of skin and mesenchymal tissues where it facilitates cell migration during wound healing, inflammation, and embryonic morphogenesis. Both during normal tissue homeostasis and particularly after tissue injury, HA is mobilized from these sites through lymphatic vessels to the lymph nodes where it is degraded before entering the circulation for rapid uptake by the liver. Currently, however, the identities of HA binding molecules which control this pathway are unknown. Here we describe the first such molecule, LYVE-1, which we have identified as a major receptor for HA on the lymph vessel wall. The deduced amino acid sequence of LYVE-1 predicts a 322-residue type I integral membrane polypeptide 41% similar to the CD44 HA receptor with a 212-residue extracellular domain containing a single Link module the prototypic HA binding domain of the Link protein superfamily. Like CD44, the LYVE-1 molecule binds both soluble and immobilized HA. However, unlike CD44, the LYVE-1 molecule colocalizes with HA on the luminal face of the lymph vessel wall and is completely absent from blood vessels. Hence, LYVE-1 is the first lymph-specific HA receptor to be characterized and is a uniquely powerful marker for lymph vessels themselves.

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Immunofluorescent  staining of LYVE-1 in lymphatic  vessels and colocalization with  HA. Sections of human small intestine were double stained for  LYVE-1, CD44, the vascular endothelial molecules CD34 and  vWF, and for HA, by indirect  immunofluorescence microscopy, using Texas red or fluorescein-conjugated secondary antibodies or bHABC as described  in Materials and Methods. The  antibody combinations were as  follows: A, LYVE-1 (red) and  CD34 (green); B, LYVE-1 (red)  and vWF (green); C, LYVE-1  (red) and CD44 (green); and D  and E, LYVE-1 (red) and  bHABC (green). Arrows depict  small blood vessels containing  erythrocytes (pale yellow/ green), which are LYVE-1-ve.  Lymph vessel endothelium positive for both LYVE-1 and HA  in D and E is stained yellow/orange. Bar, 50 μm.
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Figure 9: Immunofluorescent staining of LYVE-1 in lymphatic vessels and colocalization with HA. Sections of human small intestine were double stained for LYVE-1, CD44, the vascular endothelial molecules CD34 and vWF, and for HA, by indirect immunofluorescence microscopy, using Texas red or fluorescein-conjugated secondary antibodies or bHABC as described in Materials and Methods. The antibody combinations were as follows: A, LYVE-1 (red) and CD34 (green); B, LYVE-1 (red) and vWF (green); C, LYVE-1 (red) and CD44 (green); and D and E, LYVE-1 (red) and bHABC (green). Arrows depict small blood vessels containing erythrocytes (pale yellow/ green), which are LYVE-1-ve. Lymph vessel endothelium positive for both LYVE-1 and HA in D and E is stained yellow/orange. Bar, 50 μm.

Mentions: Immunoperoxidase staining of human tissues with the polyclonal LYVE-1 serum revealed an unusual and highly restricted expression pattern, as shown in Fig. 8 and summarized in Table I. Intriguingly the expression of LYVE-1 was mostly confined to endothelial cells lining vessels of the lymphatic system. The only other sites of prominent LYVE-1 expression were sinusoidal endothelial cells within the spleen and placental syncytiotrophoblasts (Table I and Fig. 8 k). In contrast with the expression pattern of CD44, no LYVE-1 was detected on lymphocytes or other hematopoietic cells within lymphoid organs such as the tonsil and thymus (Table I and Fig. 8 l). Localization of LYVE-1 within the lymphatics was most strikingly visible among the draining lymphatic vessels within the submucosae, underlying the intestinal crypts in colon and small intestine (Fig. 8, a–d). Many small lymph vessels, including some containing large numbers of lymphocytes, showed intense endothelial staining. High levels of LYVE-1 expression were also apparent on the lacteal vessels draining individual intestinal villi, on subdermal lymphatic vessels within the skin, and on lymphatic vessels within peripheral nodes (Fig. 8, e–j). A similar expression pattern was found in other tissues displaying prominent lymphatics, particularly the appendix and stomach (see Table I). In contrast, no LYVE-1 was detected on blood vessels within any of the tissues tested. Interestingly, LYVE-1 protein expression was detected on the surface of cultured HUVEC (Fig. 8 m), confirming the detection of LYVE-1 mRNA by RT-PCR (see above). This is most likely to be the result of culture induced de-differentiation as no vascular staining was observed in tissue sections of umbilical vein (not shown). The selectivity for lymphatic expression is underlined by a comparison of the two vessel types in sections of salivary gland (Fig. 8 g), where blood vessels that are clearly distinguishable by their erythrocyte content (visible through weak nonspecific staining with rabbit IgG) are negative for LYVE-1, while the adjacent empty lymph vessels stain intensely. This distinction was confirmed by double immunofluorescence staining of small intestine sections (Fig. 9) with antibodies to LYVE-1 and to the vascular endothelial markers CD34 (sgp90) and vWF (Factor VIII–related molecule). LYVE-1 staining was mutually exclusive with both of the other markers, and was confined to irregularly shaped vessels within the submucosae that were devoid of erythrocytes. Furthermore, double immunofluorescence staining revealed little or no CD44 expression on lymphatic vessels, indicating that LYVE-1 is likely to be the major receptor for HA on lymphatic endothelial cells (Fig. 9).


LYVE-1, a new homologue of the CD44 glycoprotein, is a lymph-specific receptor for hyaluronan.

Banerji S, Ni J, Wang SX, Clasper S, Su J, Tammi R, Jones M, Jackson DG - J. Cell Biol. (1999)

Immunofluorescent  staining of LYVE-1 in lymphatic  vessels and colocalization with  HA. Sections of human small intestine were double stained for  LYVE-1, CD44, the vascular endothelial molecules CD34 and  vWF, and for HA, by indirect  immunofluorescence microscopy, using Texas red or fluorescein-conjugated secondary antibodies or bHABC as described  in Materials and Methods. The  antibody combinations were as  follows: A, LYVE-1 (red) and  CD34 (green); B, LYVE-1 (red)  and vWF (green); C, LYVE-1  (red) and CD44 (green); and D  and E, LYVE-1 (red) and  bHABC (green). Arrows depict  small blood vessels containing  erythrocytes (pale yellow/ green), which are LYVE-1-ve.  Lymph vessel endothelium positive for both LYVE-1 and HA  in D and E is stained yellow/orange. Bar, 50 μm.
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Related In: Results  -  Collection

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Figure 9: Immunofluorescent staining of LYVE-1 in lymphatic vessels and colocalization with HA. Sections of human small intestine were double stained for LYVE-1, CD44, the vascular endothelial molecules CD34 and vWF, and for HA, by indirect immunofluorescence microscopy, using Texas red or fluorescein-conjugated secondary antibodies or bHABC as described in Materials and Methods. The antibody combinations were as follows: A, LYVE-1 (red) and CD34 (green); B, LYVE-1 (red) and vWF (green); C, LYVE-1 (red) and CD44 (green); and D and E, LYVE-1 (red) and bHABC (green). Arrows depict small blood vessels containing erythrocytes (pale yellow/ green), which are LYVE-1-ve. Lymph vessel endothelium positive for both LYVE-1 and HA in D and E is stained yellow/orange. Bar, 50 μm.
Mentions: Immunoperoxidase staining of human tissues with the polyclonal LYVE-1 serum revealed an unusual and highly restricted expression pattern, as shown in Fig. 8 and summarized in Table I. Intriguingly the expression of LYVE-1 was mostly confined to endothelial cells lining vessels of the lymphatic system. The only other sites of prominent LYVE-1 expression were sinusoidal endothelial cells within the spleen and placental syncytiotrophoblasts (Table I and Fig. 8 k). In contrast with the expression pattern of CD44, no LYVE-1 was detected on lymphocytes or other hematopoietic cells within lymphoid organs such as the tonsil and thymus (Table I and Fig. 8 l). Localization of LYVE-1 within the lymphatics was most strikingly visible among the draining lymphatic vessels within the submucosae, underlying the intestinal crypts in colon and small intestine (Fig. 8, a–d). Many small lymph vessels, including some containing large numbers of lymphocytes, showed intense endothelial staining. High levels of LYVE-1 expression were also apparent on the lacteal vessels draining individual intestinal villi, on subdermal lymphatic vessels within the skin, and on lymphatic vessels within peripheral nodes (Fig. 8, e–j). A similar expression pattern was found in other tissues displaying prominent lymphatics, particularly the appendix and stomach (see Table I). In contrast, no LYVE-1 was detected on blood vessels within any of the tissues tested. Interestingly, LYVE-1 protein expression was detected on the surface of cultured HUVEC (Fig. 8 m), confirming the detection of LYVE-1 mRNA by RT-PCR (see above). This is most likely to be the result of culture induced de-differentiation as no vascular staining was observed in tissue sections of umbilical vein (not shown). The selectivity for lymphatic expression is underlined by a comparison of the two vessel types in sections of salivary gland (Fig. 8 g), where blood vessels that are clearly distinguishable by their erythrocyte content (visible through weak nonspecific staining with rabbit IgG) are negative for LYVE-1, while the adjacent empty lymph vessels stain intensely. This distinction was confirmed by double immunofluorescence staining of small intestine sections (Fig. 9) with antibodies to LYVE-1 and to the vascular endothelial markers CD34 (sgp90) and vWF (Factor VIII–related molecule). LYVE-1 staining was mutually exclusive with both of the other markers, and was confined to irregularly shaped vessels within the submucosae that were devoid of erythrocytes. Furthermore, double immunofluorescence staining revealed little or no CD44 expression on lymphatic vessels, indicating that LYVE-1 is likely to be the major receptor for HA on lymphatic endothelial cells (Fig. 9).

Bottom Line: Like CD44, the LYVE-1 molecule binds both soluble and immobilized HA.However, unlike CD44, the LYVE-1 molecule colocalizes with HA on the luminal face of the lymph vessel wall and is completely absent from blood vessels.Hence, LYVE-1 is the first lymph-specific HA receptor to be characterized and is a uniquely powerful marker for lymph vessels themselves.

View Article: PubMed Central - PubMed

Affiliation: University of Oxford, Molecular Immunology Group, Nuffield Department of Medicine, John Radcliff Hospital, Headington, Oxford OX3 9DU, United Kingdom.

ABSTRACT
The extracellular matrix glycosaminoglycan hyaluronan (HA) is an abundant component of skin and mesenchymal tissues where it facilitates cell migration during wound healing, inflammation, and embryonic morphogenesis. Both during normal tissue homeostasis and particularly after tissue injury, HA is mobilized from these sites through lymphatic vessels to the lymph nodes where it is degraded before entering the circulation for rapid uptake by the liver. Currently, however, the identities of HA binding molecules which control this pathway are unknown. Here we describe the first such molecule, LYVE-1, which we have identified as a major receptor for HA on the lymph vessel wall. The deduced amino acid sequence of LYVE-1 predicts a 322-residue type I integral membrane polypeptide 41% similar to the CD44 HA receptor with a 212-residue extracellular domain containing a single Link module the prototypic HA binding domain of the Link protein superfamily. Like CD44, the LYVE-1 molecule binds both soluble and immobilized HA. However, unlike CD44, the LYVE-1 molecule colocalizes with HA on the luminal face of the lymph vessel wall and is completely absent from blood vessels. Hence, LYVE-1 is the first lymph-specific HA receptor to be characterized and is a uniquely powerful marker for lymph vessels themselves.

Show MeSH
Related in: MedlinePlus