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Molecular analysis of the interaction of LCMV with its cellular receptor [alpha]-dystroglycan.

Kunz S, Sevilla N, McGavern DB, Campbell KP, Oldstone MB - J. Cell Biol. (2001)

Bottom Line: In the present study, we characterized the binding of LCMV to alpha-DG and addressed the role of alpha-DG-associated host-derived proteins in virus infection.We found that the COOH-terminal region of alpha-DG's first globular domain and the NH2-terminal region of the mucin-related structures of alpha-DG together form the binding site for LCMV.This competition of the virus with ECM molecules for receptor binding likely explains the recently found correlation between the affinity of LCMV binding to alpha-DG, tissue tropism, and pathological potential.

View Article: PubMed Central - PubMed

Affiliation: The Scripps Research Institute, Division of Virology, Department of Neuropharmacology, La Jolla, CA 92037, USA.

ABSTRACT
alpha-Dystroglycan (DG) has been identified as the cellular receptor for lymphocytic choriomeningitis virus (LCMV) and Lassa fever virus (LFV). This subunit of DG is a highly versatile cell surface molecule that provides a molecular link between the extracellular matrix (ECM) and a beta-DG transmembrane component, which interacts with the actin-based cytoskeleton. In addition, DG exhibits a complex pattern of interaction with a wide variety of ECM and cellular proteins. In the present study, we characterized the binding of LCMV to alpha-DG and addressed the role of alpha-DG-associated host-derived proteins in virus infection. We found that the COOH-terminal region of alpha-DG's first globular domain and the NH2-terminal region of the mucin-related structures of alpha-DG together form the binding site for LCMV. The virus-alpha-DG binding unlike ECM alpha-DG interactions was not dependent on divalent cations. Despite such differences in binding, LCMV and laminin-1 use, in part, an overlapping binding site on alpha-DG, and the ability of an LCMV isolate to compete with laminin-1 for receptor binding is determined by its binding affinity to alpha-DG. This competition of the virus with ECM molecules for receptor binding likely explains the recently found correlation between the affinity of LCMV binding to alpha-DG, tissue tropism, and pathological potential. LCMV strains and variants with high binding affinity to alpha-DG but not low affinity binders are able to infect CD11c+ dendritic cells, which express alpha-DG at their surface. Infection followed by dysfunction of these antigen-presenting cells contributes to immunosuppression and persistent viral infection in vivo.

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Expression pattern of laminin and CD11c and anatomic localization of viral nucleic acids of LCMV ARM53b and clone-13 in the spleen. Detection of laminin and CD11c in whole spleen sections. (a) Laminin was detected with a rabbit anti–mouse laminin-1 antibody and a FITC-labeled secondary antibody (green). CD11c was detected with a hamster anti-CD11c antibody and a rhodamine-X–conjugated secondary antibody (red). Overlapping fluorescence appears in yellow, since a rhodamine-X/FITC double filter device was used. Localization of laminin in the marginal zone of the white pulp. (b) Laminin was stained with rabbit anti–laminin-1 antibody using an HRP-conjugated secondary antibody and DAB as a chromophore. Sections were counter stained with hematoxylin and eosin. Laminin staining of a single white pulp area. (c and d) Laminin was detected with a rabbit anti–laminin-1 antibody and a Texas red–conjugated secondary antibody (c). Secondary antibody is shown only in d. Detection of viral nucleic acids of LCMV clone-13 (e) and ARM53b (f) within the spleen 3 d after infection. Spleen sections from mice infected 3 d before with 2 × 106 pfu (i.v.) clone-13 (e), ARM53b (f), or mock infection (g) were examined by in situ hybridization using a digoxigenin-labeled riboprobe specific for LCMV-NP. MZ, marginal zone; RP, red pulp; WP, white pulp. Bars: (a) 200 μm; (b–g) 50 μm.
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fig8: Expression pattern of laminin and CD11c and anatomic localization of viral nucleic acids of LCMV ARM53b and clone-13 in the spleen. Detection of laminin and CD11c in whole spleen sections. (a) Laminin was detected with a rabbit anti–mouse laminin-1 antibody and a FITC-labeled secondary antibody (green). CD11c was detected with a hamster anti-CD11c antibody and a rhodamine-X–conjugated secondary antibody (red). Overlapping fluorescence appears in yellow, since a rhodamine-X/FITC double filter device was used. Localization of laminin in the marginal zone of the white pulp. (b) Laminin was stained with rabbit anti–laminin-1 antibody using an HRP-conjugated secondary antibody and DAB as a chromophore. Sections were counter stained with hematoxylin and eosin. Laminin staining of a single white pulp area. (c and d) Laminin was detected with a rabbit anti–laminin-1 antibody and a Texas red–conjugated secondary antibody (c). Secondary antibody is shown only in d. Detection of viral nucleic acids of LCMV clone-13 (e) and ARM53b (f) within the spleen 3 d after infection. Spleen sections from mice infected 3 d before with 2 × 106 pfu (i.v.) clone-13 (e), ARM53b (f), or mock infection (g) were examined by in situ hybridization using a digoxigenin-labeled riboprobe specific for LCMV-NP. MZ, marginal zone; RP, red pulp; WP, white pulp. Bars: (a) 200 μm; (b–g) 50 μm.

Mentions: Since earlier studies revealed a strikingly different tropism between LCMV ARM53b and clone-13 in spleen (Borrow et al., 1995; Sevilla et al., 2000), we compared the expression of laminin with the expression of the DC marker CD11c and the infection pattern of ARM53b and clone-13 in this tissue. Immunohistochemical analysis revealed intense laminin expression in the marginal zone at the periphery of the white pulp and surrounding blood vessels (Fig. 8, a, b, and c) with comparatively minimal staining in the surrounding red pulp. CD11c+ cell clusters were detected mainly in these laminin-rich areas as illustrated by the extensive overlap between laminin and CD11c staining (Fig. 8 a). Concurrently, the anatomic distribution of ARM53b and clone-13 was examined 3 d after infection with 2 × 106 plaque-forming units (pfu) of virus (i.v.) using a digoxigenin-labeled probe to LCMV-NP and in situ hybridization (Fig. 8, e and f) and compared with the localization of laminin (Fig. 8, b and c). Consistent with previous studies (Borrow et al., 1995; Fig. 4), LCMV clone-13 localized predominantly to cells in the marginal zone and white pulp (Fig. 8 e), whereas ARM53b localized primarily to cells in the red pulp (Fig. 8 f). Infection of cells in the marginal zone of the white pulp with clone-13 colocalized with the expression of laminin and CD11c. In contrast, there was a disassociation between infection with ARM53b, which was limited to the surrounding red pulp with the expression of laminin.


Molecular analysis of the interaction of LCMV with its cellular receptor [alpha]-dystroglycan.

Kunz S, Sevilla N, McGavern DB, Campbell KP, Oldstone MB - J. Cell Biol. (2001)

Expression pattern of laminin and CD11c and anatomic localization of viral nucleic acids of LCMV ARM53b and clone-13 in the spleen. Detection of laminin and CD11c in whole spleen sections. (a) Laminin was detected with a rabbit anti–mouse laminin-1 antibody and a FITC-labeled secondary antibody (green). CD11c was detected with a hamster anti-CD11c antibody and a rhodamine-X–conjugated secondary antibody (red). Overlapping fluorescence appears in yellow, since a rhodamine-X/FITC double filter device was used. Localization of laminin in the marginal zone of the white pulp. (b) Laminin was stained with rabbit anti–laminin-1 antibody using an HRP-conjugated secondary antibody and DAB as a chromophore. Sections were counter stained with hematoxylin and eosin. Laminin staining of a single white pulp area. (c and d) Laminin was detected with a rabbit anti–laminin-1 antibody and a Texas red–conjugated secondary antibody (c). Secondary antibody is shown only in d. Detection of viral nucleic acids of LCMV clone-13 (e) and ARM53b (f) within the spleen 3 d after infection. Spleen sections from mice infected 3 d before with 2 × 106 pfu (i.v.) clone-13 (e), ARM53b (f), or mock infection (g) were examined by in situ hybridization using a digoxigenin-labeled riboprobe specific for LCMV-NP. MZ, marginal zone; RP, red pulp; WP, white pulp. Bars: (a) 200 μm; (b–g) 50 μm.
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fig8: Expression pattern of laminin and CD11c and anatomic localization of viral nucleic acids of LCMV ARM53b and clone-13 in the spleen. Detection of laminin and CD11c in whole spleen sections. (a) Laminin was detected with a rabbit anti–mouse laminin-1 antibody and a FITC-labeled secondary antibody (green). CD11c was detected with a hamster anti-CD11c antibody and a rhodamine-X–conjugated secondary antibody (red). Overlapping fluorescence appears in yellow, since a rhodamine-X/FITC double filter device was used. Localization of laminin in the marginal zone of the white pulp. (b) Laminin was stained with rabbit anti–laminin-1 antibody using an HRP-conjugated secondary antibody and DAB as a chromophore. Sections were counter stained with hematoxylin and eosin. Laminin staining of a single white pulp area. (c and d) Laminin was detected with a rabbit anti–laminin-1 antibody and a Texas red–conjugated secondary antibody (c). Secondary antibody is shown only in d. Detection of viral nucleic acids of LCMV clone-13 (e) and ARM53b (f) within the spleen 3 d after infection. Spleen sections from mice infected 3 d before with 2 × 106 pfu (i.v.) clone-13 (e), ARM53b (f), or mock infection (g) were examined by in situ hybridization using a digoxigenin-labeled riboprobe specific for LCMV-NP. MZ, marginal zone; RP, red pulp; WP, white pulp. Bars: (a) 200 μm; (b–g) 50 μm.
Mentions: Since earlier studies revealed a strikingly different tropism between LCMV ARM53b and clone-13 in spleen (Borrow et al., 1995; Sevilla et al., 2000), we compared the expression of laminin with the expression of the DC marker CD11c and the infection pattern of ARM53b and clone-13 in this tissue. Immunohistochemical analysis revealed intense laminin expression in the marginal zone at the periphery of the white pulp and surrounding blood vessels (Fig. 8, a, b, and c) with comparatively minimal staining in the surrounding red pulp. CD11c+ cell clusters were detected mainly in these laminin-rich areas as illustrated by the extensive overlap between laminin and CD11c staining (Fig. 8 a). Concurrently, the anatomic distribution of ARM53b and clone-13 was examined 3 d after infection with 2 × 106 plaque-forming units (pfu) of virus (i.v.) using a digoxigenin-labeled probe to LCMV-NP and in situ hybridization (Fig. 8, e and f) and compared with the localization of laminin (Fig. 8, b and c). Consistent with previous studies (Borrow et al., 1995; Fig. 4), LCMV clone-13 localized predominantly to cells in the marginal zone and white pulp (Fig. 8 e), whereas ARM53b localized primarily to cells in the red pulp (Fig. 8 f). Infection of cells in the marginal zone of the white pulp with clone-13 colocalized with the expression of laminin and CD11c. In contrast, there was a disassociation between infection with ARM53b, which was limited to the surrounding red pulp with the expression of laminin.

Bottom Line: In the present study, we characterized the binding of LCMV to alpha-DG and addressed the role of alpha-DG-associated host-derived proteins in virus infection.We found that the COOH-terminal region of alpha-DG's first globular domain and the NH2-terminal region of the mucin-related structures of alpha-DG together form the binding site for LCMV.This competition of the virus with ECM molecules for receptor binding likely explains the recently found correlation between the affinity of LCMV binding to alpha-DG, tissue tropism, and pathological potential.

View Article: PubMed Central - PubMed

Affiliation: The Scripps Research Institute, Division of Virology, Department of Neuropharmacology, La Jolla, CA 92037, USA.

ABSTRACT
alpha-Dystroglycan (DG) has been identified as the cellular receptor for lymphocytic choriomeningitis virus (LCMV) and Lassa fever virus (LFV). This subunit of DG is a highly versatile cell surface molecule that provides a molecular link between the extracellular matrix (ECM) and a beta-DG transmembrane component, which interacts with the actin-based cytoskeleton. In addition, DG exhibits a complex pattern of interaction with a wide variety of ECM and cellular proteins. In the present study, we characterized the binding of LCMV to alpha-DG and addressed the role of alpha-DG-associated host-derived proteins in virus infection. We found that the COOH-terminal region of alpha-DG's first globular domain and the NH2-terminal region of the mucin-related structures of alpha-DG together form the binding site for LCMV. The virus-alpha-DG binding unlike ECM alpha-DG interactions was not dependent on divalent cations. Despite such differences in binding, LCMV and laminin-1 use, in part, an overlapping binding site on alpha-DG, and the ability of an LCMV isolate to compete with laminin-1 for receptor binding is determined by its binding affinity to alpha-DG. This competition of the virus with ECM molecules for receptor binding likely explains the recently found correlation between the affinity of LCMV binding to alpha-DG, tissue tropism, and pathological potential. LCMV strains and variants with high binding affinity to alpha-DG but not low affinity binders are able to infect CD11c+ dendritic cells, which express alpha-DG at their surface. Infection followed by dysfunction of these antigen-presenting cells contributes to immunosuppression and persistent viral infection in vivo.

Show MeSH
Related in: MedlinePlus