An Ultrastructural Investigation of Cytomegalovirus Replication in Murine Hepatocytes

Papadimitriou, John; Shellam, Geoffrey; Robertson, Terry

doi:10.1099/0022-1317-65-11-1979

Cited by 18 publications

(25 citation statements)

References 28 publications

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“…1C) (49). Second, MCMV directly infects hepatocytes, whereas LCMV (the Armstrong strain in particular) primarily causes infection of Kuppfer cells and the periportal areas (12,39). Therefore, different chemokine requirements may exist for cell infiltration and virus control in these two viral systems.…”

Section: Discussionmentioning

confidence: 99%

“…7 days earlier were incubated in vitro with or without LCMV-derived peptide (gp [33][34][35][36][37][38][39][40][41] ). At the indicated time points, monensin was added to the cultures, and CD8 ϩ cells were analyzed for cytokine production by intracellular staining 1 h later; cells from uninfected mice were included as a control.…”

Section: Fig 8 Elimination Of Cd8mentioning

confidence: 99%

“…We have recently taken the rational approach and attempted to unravel redundancy within the chemokine system by gener-b /gp [33][34][35][36][37][38][39][40][41] or H-2D b /np [396][397][398][399][400][401][402][403][404] dextramers kindly provided by Dako (Glostrup, Denmark).…”

mentioning

confidence: 99%

“…Surface staining of cells for flow cytometry was performed according to standard laboratory procedure (2). For the evaluation of cytokine production by LCMV-specific CD8 ϩ T cells, splenocytes were incubated in vitro at 37°C in 5% CO 2 with or without gp [33][34][35][36][37][38][39][40][41] peptide (0.1 g/ml) in the presence of monensin (3 M; Sigma Chemicals Co., St. Louis, MO), and murine recombinant interleukin-2 (10 U/well; R&D Systems Europe Ltd., Abingdon, United Kingdom). After incubation, cells were surface stained, washed, and permeabilized using 0.5% saponin.…”

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confidence: 99%

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CCR5 and CXCR3 Are Dispensable for Liver Infiltration, but CCR5 Protects against Virus-Induced T-Cell-Mediated Hepatic Steatosis

Holst

Ørskov

Qvortrup

et al. 2007

J Virol

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CCR5 and CXCR3 are important molecules in regulating the migration of activated lymphocytes. Thus, the majority of tissue-infiltrating T cells found in the context of autoimmune conditions and viral infections express CCR5 and CXCR3, and the principal chemokine ligands are expressed within inflamed tissues. Accordingly, intervention studies have pointed to nonredundant roles of these receptors in models of allograft rejection, viral infection, and autoimmunity. In spite of this, considerable controversy exists, with many studies failing to support a role for CCR5 or CXCR3 in disease pathogenesis. One possible explanation is that different chemokine receptors may take over in the absence of any individual receptor, thus rendering individual receptors redundant. We have attempted to address this issue by analyzing CCR5 ؊/؊ , CXCR3 ؊/؊ , and CCR5/CXCR3 ؊/؊ mice with regard to virus-induced liver inflammation, generation and recruitment of effector cells, virus control, and immunopathology. Our results indicate that CCR5 and CXCR3 are largely dispensable for tissue infiltration and virus control. In contrast, the T-cell response is accelerated in CCR5 ؊/؊ and CCR5/CXCR3 ؊/؊ mice and the absence of CCR5 is associated with the induction of CD8 ؉ T-cell-mediated immunopathology consisting of marked hepatic microvesicular steatosis.Chemokines play a key role in virus-induced inflammation and virus clearance, acting as regulators of leukocyte trafficking (18,49). Among the chemokine receptors, CCR5 has drawn considerable attention as the main coreceptor for human immunodeficiency virus and CXCR3 is a promising target for antiinflammatory therapeutics (41, 52). Notably, both receptors have been associated with type 1 cytokine T-cell responses and the majority of tissue-infiltrating T cells in several human inflammatory diseases are found to coexpress these receptors (7,42,48). During viral infection, CCR5 and CXCR3 are expressed on NK cells and activated CD4 ϩ and CD8 ϩ T cells and the ligands are generally expressed in virus-infected tissues (30,38,50). Further support for the important role of CCR5 and CXCR3 comes from the existence of virally encoded chemokine receptor antagonists targeting these receptors (23,27). In this context, it is not surprising that CXCR3 or its ligands have been experimentally demonstrated to be important for immune-mediated virus control in the central nervous system (CNS), the lungs, the ovaries, and the islet of Langerhans in the pancreas (9,15,20,26,51). In contrast, despite the suggestive expression patterns, several studies have indicated that CCR5 or CCR5 ligands are less important in antiviral immunity (30, 37). The positive findings suggest a role for CCR5 in macrophage and CD4 ϩ T-cell recruitment, but the receptor appears not to be required for CD8 ϩ T-cell migration except in young mice, in which the immune system has not yet reached full maturity (3, 17). The reason for this difference between the two receptors could lie in a difference in biological redundancy.Thus, several CCR5 ...

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Section: Discussionmentioning

confidence: 99%

Section: Fig 8 Elimination Of Cd8mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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CCR5 and CXCR3 Are Dispensable for Liver Infiltration, but CCR5 Protects against Virus-Induced T-Cell-Mediated Hepatic Steatosis

Holst

Ørskov

Qvortrup

et al. 2007

J Virol

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“…These structures were also observed in confocal images corresponding to the midsections of nuclei, suggesting that they might correspond to the extensive invaginations of the inner nuclear membrane (INM) described in ultrastructural studies and in immunofluorescence staining analyses of human-and mouse-CMV-infected cells at late times p.i. (6,7,14,24,35,50,62,67). To determine if this was the case, images were acquired at three different focal planes of a single nucleus, corresponding to the top (Fig.…”

Section: Fig 3 Rascal Intracellular Localization Mock-infected (A mentioning

confidence: 99%

RASCAL Is a New Human Cytomegalovirus-Encoded Protein That Localizes to the Nuclear Lamina and in Cytoplasmic Vesicles at Late Times Postinfection

Miller

Furlong

Pennell

et al. 2010

J Virol

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The products of numerous open reading frames (ORFs) present in the genome of human cytomegalovirus (CMV) have not been characterized. Here, we describe the identification of a new CMV protein localizing to the nuclear envelope and in cytoplasmic vesicles at late times postinfection. Based on this distinctive localization pattern, we called this new protein nuclear rim-associated cytomegaloviral protein, or RASCAL. Two RASCAL isoforms exist, a short version of 97 amino acids encoded by the majority of CMV strains and a longer version of 176 amino acids encoded by the Towne, Toledo, HAN20, and HAN38 strains. Both isoforms colocalize with lamin B in deep intranuclear invaginations of the inner nuclear membrane (INM) and in novel cytoplasmic vesicular structures possibly derived from the nuclear envelope. INM infoldings have been previously described as sites of nucleocapsid egress, which is mediated by the localized disruption of the nuclear lamina, promoted by the activities of viral and cellular kinases recruited by the lamina-associated proteins UL50 and UL53. RASCAL accumulation at the nuclear membrane required the presence of UL50 but not of UL53. RASCAL and UL50 also appeared to specifically interact, suggesting that RASCAL is a new component of the nuclear egress complex (NEC) and possibly involved in mediating nucleocapsid egress from the nucleus. Finally, the presence of RASCAL within cytoplasmic vesicles raises the intriguing possibility that this protein might participate in additional steps of virion maturation occurring after capsid release from the nucleus.

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Morphological and Functional Changes During Cytomegalovirus Replication in Murine Macrophages

Bruggen

Price

Robertson

et al. 1989

Journal of Leukocyte Biology

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Structural and functional changes were studied in murine peritoneal macrophages infected with murine cytomegalovirus by using centrifugal enhancement to achieve a high-level (> 90%) pulsed infection. During 3 d of culture the infected cells became enlarged and rounded with smooth surface contours. Transmission electron microscopy demonstrated various stages of viral maturation in the nucleus and cytoplasm. Intracellular organization was generally retained, apart from the development of large, irregular, intracytoplasmic vacuoles, in which enveloped virions and cell debris accumulated. The infected macrophages lost most surface markers tested (F4/80, Mac-1, FcR, and the receptor for gluteraldehyde-fixed sheep red blood cells), but H-2 expression was increased. Moreover, ingestion of colloidal gold or horseradish peroxidase was depressed, and the levels of acid phosphatase activity, lymphocytostatic activity, and interleukin 1 production were also decreased. The latter may explain the observed loss of accessory cell function.

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An Ultrastructural Investigation of Cytomegalovirus Replication in Murine Hepatocytes

Cited by 18 publications

References 28 publications

CCR5 and CXCR3 Are Dispensable for Liver Infiltration, but CCR5 Protects against Virus-Induced T-Cell-Mediated Hepatic Steatosis

CCR5 and CXCR3 Are Dispensable for Liver Infiltration, but CCR5 Protects against Virus-Induced T-Cell-Mediated Hepatic Steatosis

RASCAL Is a New Human Cytomegalovirus-Encoded Protein That Localizes to the Nuclear Lamina and in Cytoplasmic Vesicles at Late Times Postinfection

Morphological and Functional Changes During Cytomegalovirus Replication in Murine Macrophages

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