Susceptibility of Immature and Mature Langerhans Cell-Type Dendritic Cells to Infection and Immunomodulation by Human Cytomegalovirus

Hertel, Laura; Lacaille, Vashti; Strobl, Herbert; Mellins, Elizabeth; Mocarski, Edward S.

doi:10.1128/jvi.77.13.7563-7574.2003

Cited by 118 publications

(164 citation statements)

References 78 publications

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“…Therefore, there has been increasing interest in studying HCMV infection of endothelial cells (EC) and monocyte-derived cells, which are assumed to contribute greatly to the haematogenous spread of the virus Hertel et al, 2003;Jarvis & Nelson, 2002;Sinzger & Jahn, 1996). This has been facilitated by the introduction of EC-propagated HCMV strains that have preserved their natural broad cell tropism in cell culture.…”

Section: Introductionmentioning

confidence: 99%

Cloning and sequencing of a highly productive, endotheliotropic virus strain derived from human cytomegalovirus TB40/E

Sinzger

Hahn

Digel

et al. 2008

Journal of General Virology

358

355

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Human cytomegalovirus (HCMV) strain TB40/E, replicates efficiently, exhibits a broad cell tropism and is widely used for infection of endothelial cells and monocyte-derived cells yet has not been available in a phenotypically homogeneous form compatible with genetic analysis. To overcome this problem, we cloned the TB40/E strain into a bacterial artificial chromosome (BAC) vector. Both highly endotheliotropic and poorly endotheliotropic virus clones, representing three distinct restriction fragment patterns, were reconstituted after transfection of BAC clones derived from previously plaque-purified strain TB40/E. For one of the highly endotheliotropic clones, TB40-BAC4, we provide the genome sequence. Two BACs with identical restriction fragment patterns but different cell tropism were further analysed in the UL128-UL131A gene region. Sequence analysis revealed one coding-relevant adenine insertion at position 332 of UL128 in the BAC of the poorly endotheliotropic virus, which caused a frameshift in the C-terminal part of the coding sequence. Removal of this insertion by markerless mutagenesis restored the highly endotheliotropic phenotype, indicating that the loss of endothelial cell tropism was caused by this insertion. In conclusion, HCMV strain TB40/E, which combines the high endothelial cell tropism of a clinical isolate with the high titre growth of a cell culture adapted strain, is now available as a BAC clone suitable for genetic engineering. The results also suggest BAC cloning as a suitable method for selection of genetically defined virus clones.

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Section: Introductionmentioning

confidence: 99%

Cloning and sequencing of a highly productive, endotheliotropic virus strain derived from human cytomegalovirus TB40/E

Sinzger

Hahn

Digel

et al. 2008

Journal of General Virology

358

355

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“…DC are specialized antigen-presenting cells which, in common with several other cell types, such as endothelial cells, can be experimentally infected with HCMV in vitro (18,19). There are at least two types of myeloid lineage-derived DC: the Langerhans DC that are located in the epithelium (and could, therefore, be one of the first cell types to encounter HCMV after infection) and the interstitial DC that reside in deeper-lying tissues (reviewed in ref.…”

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confidence: 99%

Latency, chromatin remodeling, and reactivation of human cytomegalovirus in the dendritic cells of healthy carriers

Reeves

MacAry

Lehner

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

320

412

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Human cytomegalovirus (HCMV) persists as a subclinical, lifelong infection in the normal human host, but reactivation from latency in immunocompromised subjects results in serious disease. Latency and reactivation are defining characteristics of the herpesviruses and are key to understanding their biology; however, the precise cellular sites in which HCMV is carried and the mechanisms regulating its latency and reactivation during natural infection remain poorly understood. Here we present evidence, based entirely on direct analysis of material isolated from healthy virus carriers, to show that myeloid dendritic cell (DC) progenitors are sites of HCMV latency and that their ex vivo differentiation to a mature DC phenotype is linked with reactivation of infectious virus resulting from differentiation-dependent chromatin remodeling of the viral major immediate-early promoter. Thus, myeloid DC progenitors are a site of HCMV latency during natural persistence, and there is a critical linkage between their differentiation to DC and transcriptional reactivation of latent virus, which is likely to play an important role in the pathogenesis of HCMV infection. P rimary infection of healthy individuals with human cytomegalovirus (HCMV) is often asymptomatic and results in lifelong persistence in the host, a characteristic of all herpes viruses. However, primary infection and reactivation of latent HCMV causes serious disease in immunosuppressed transplant recipients and in advanced HIV infection (1-3). Transfusionmediated HCMV disease can be prevented by leukocyte depletion (4) of blood, but infectious virus cannot be detected in the blood of healthy carriers, suggesting that HCMV is transmitted as latent virus in the peripheral blood leukocyte population. Accumulating evidence has shown that HCMV is carried latently in mononuclear cells of the myeloid lineage during lifelong latency in naturally infected individuals (5-8). Differentiation of monocytes to macrophages in vitro is reported to induce immediate-early (IE) lytic gene expression from latent virus (9), and groups have intermittently reported that infectious virus can be recovered after differentiation of monocytes to macrophages through explant culture (10) and, more recently, by allogeneic T cell stimulation (11), suggesting that reactivation of latent virus is associated with both the differentiation and activation state of myeloid cells.The major IE genes of HCMV, driven by the viral major IE promoter͞enhancer (MIEP), are the two most abundantly transcribed genes at IE times of virus lytic infection (12), and their proteins play a critical role in control of viral early and late gene expression (for review, see ref. 12). Consistent with the differentiation-dependent induction of IE gene expression observed above, there is a clear correlation between the differentiation state of the cell and the regulation of HCMV IE gene expression in vitro. Thus, in transfection assays, the MIEP is transcriptionally repressed in undifferentiated but transcriptionally active ...

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“…Human CMV strains AD169varATCC, RV798, TB40/E and Toledo were kind gifts from E. S. Mocarski and were originally obtained from the American Type Culture Collection (ATCC), T. Jones (Wyeth Research, Pearl River, NY, USA), S. Plotkin (Aventis Pasteur, Swiftwater, PA, USA) and C. Sinzger (University of Tübingen, Tübingen, Germany), respectively. Propagation, purification and titre determination of all strains were performed as described previously (Hertel et al, 2003). Tumour necrosis factor (TNF)-a was purchased from PeproTech and was resuspended in water.…”

Section: Methodsmentioning

confidence: 99%

“…Sinzger (University of Tübingen, Tübingen, Germany), respectively. Propagation, purification and titre determination of all strains were performed as described previously (Hertel et al, 2003). Tumour necrosis factor (TNF)-a was purchased from PeproTech and was resuspended in water.…”

mentioning

confidence: 99%

Human cytomegalovirus US9 protein contains an N-terminal signal sequence and a C-terminal mitochondrial localization domain, and does not alter cellular sensitivity to apoptosis

Mandic

Miller

Coulter

et al. 2009

Journal of General Virology

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The human cytomegalovirus (CMV) US2-US11 genomic region contains a cluster of genes whose products interfere with antigen presentation by the major histocompatibility complex (MHC) proteins. Although included in this cluster, the US9 gene encodes a glycoprotein that does not affect MHC activity and whose function is still largely uncharacterized. An in silico analysis of the US9 amino-acid sequence uncovered the presence of an N-terminal signal sequence (SS) and a C-terminal transmembrane domain containing the specific hallmarks of known mitochondrial localization sequences (MLS). Expression of full-length US9 and of US9 deletion mutants fused to GFP revealed that the N-terminal SS mediates US9 targeting to the endoplasmic reticulum (ER) and that the C-terminal MLS is both necessary and sufficient to direct US9 to mitochondria in the absence of a functional SS. This dual localization suggested a possible role for US9 in protection from apoptosis triggered by ER-to-mitochondria signalling. Fibroblasts infected with the US2-US11 deletion mutant virus RV798 or with the parental strain AD169varATCC were equally susceptible to death triggered by exposure to tumour necrosis factor (TNF)-a, tunicamycin, thapsigargin, brefeldin A, lonidamine and carbonyl cyanide m-chloro phenyl hydrazone, but were 1.6-fold more sensitive to apoptosis induced by hygromycin B. Expression of US9 in human embryonic kidney 293T cells or in fibroblasts, however, did not protect cells from hygromycin B-mediated death. Together, these results classify US9 as the first CMV-encoded protein to contain an N-terminal SS and a C-terminal MLS, and suggest a completely novel role for this protein during infection. INTRODUCTIONHuman cytomegalovirus (CMV) is a very prevalent betaherpesvirus that poses serious medical concerns in immunocompromised individuals . The ability to evade or modulate host immune responses, coupled with the capacity to establish latency after primary infection, is at the basis of the tremendous success of CMV as a human pathogen.The US2-US11 genomic region has been shown to be dispensable for viral replication in primary human fibroblasts (HF) (Jones & Muzithras, 1992;Kollert-Jons et al., 1991), and to encode five endoplasmic reticulum (ER)-resident glycoproteins, US2, US3, US6, US10 and US11, that disrupt antigen presentation by the major histocompatibility complex (MHC) class I and class II proteins (Lin et al., 2007). Despite sharing some very limited homology with US6, US10 and US11, US9 does not bind to or reduce MHC cell surface levels (Hegde et al., 2002(Hegde et al., , 2006Pereira et al., 1995). US9 was originally described as a member of the US6 family of proteins, which includes the products of the US6-US11 genes. These proteins contain markedly hydrophobic sequences at both the N-and C-termini, and were predicted to insert into cellular membranes (Weston & Barrell, 1986). US9 intracellular localization was initially examined in nonpolarized Madin-Darby canine kidney cells constitutively expressing a C-terminal-tagged ...

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Susceptibility of Immature and Mature Langerhans Cell-Type Dendritic Cells to Infection and Immunomodulation by Human Cytomegalovirus

Abstract: Human cytomegalovirus (CMV) infection initiates in mucosal epithelia

Cited by 118 publications

References 78 publications

Cloning and sequencing of a highly productive, endotheliotropic virus strain derived from human cytomegalovirus TB40/E

Cloning and sequencing of a highly productive, endotheliotropic virus strain derived from human cytomegalovirus TB40/E

Latency, chromatin remodeling, and reactivation of human cytomegalovirus in the dendritic cells of healthy carriers

Human cytomegalovirus US9 protein contains an N-terminal signal sequence and a C-terminal mitochondrial localization domain, and does not alter cellular sensitivity to apoptosis

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