Human Cytomegalovirus Replicates Abortively in Polymorphonuclear Leukocytes after Transfer from Infected Endothelial Cells via Transient Microfusion Events

Gerna, Giuseppe; Percivalle, Elena; Baldanti, Fausto; Sozzani, Silvano; Lanzarini, P.; Genini, Emilia; Lilleri, Daniele; Revello, Maria Grazia

doi:10.1128/jvi.74.12.5629-5638.2000

Cited by 148 publications

(181 citation statements)

References 38 publications

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“…Recent reports have demonstrated that ICAM-1-mediated cell interactions are essential for microfusion events taking place between polymorphonuclear leukocytes (PMNLs) and infected endothelial cells (Gerna et al, 2000(Gerna et al, , 2002. Virus is delivered via these fusion events from the endothelium to transiently adherent PMNLs, which are implicated in the subsequent haematogenous spreading of HCMV.…”

Section: Discussionmentioning

confidence: 99%

“…Transmission could be prevented by inhibition of the contact between the two cell populations and by the use of monoclonal antibodies to CD18 and ICAM-1. Therefore, ICAM-1 upregulation by HCMV is a crucial determinant of viral dissemination and is even proposed as a surrogate marker of pathogenicity as well as attenuation of HCMV strains (Gerna et al, 2000;Knight et al, 2000).…”

Section: Discussionmentioning

confidence: 99%

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Synergistic induction of intercellular adhesion molecule-1 by the human cytomegalovirus transactivators IE2p86 and pp71 is mediated via an Sp1-binding site

Kronschnabl¹,

Stamminger²

2003

Journal of General Virology

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Human cytomegalovirus (HCMV) infection of transplant recipients is frequently associated with allograft vasculopathy and rejection. One potential mechanism is vascular injury from HCMVtriggered, immunologically mediated processes. HCMV infection has been shown to increase the expression of intercellular adhesion molecule-1 (ICAM-1). The objective of this study was to determine the molecular basis of HCMV-enhanced ICAM-1 gene expression. Transient transfection experiments identified the IE2p86 protein as a potent activator of the ICAM-1 promoter. The tegument protein pp71 showed a strong synergistic effect on IE2p86-mediated ICAM-1 promoter activation. Mutagenesis experiments defined a DNA element from 2110 to 242 relative to the transcription start site as responsive for IE2p86. Further point mutations within this DNA element identified an Sp1-binding site that was essential for strong synergistic activation by IE2p86 and pp71. To confirm the activation of ICAM-1 gene expression, human fibroblasts (HFF) as well as endothelial cells (HUVEC) were infected with recombinant IE2p86-and pp71-expressing baculoviruses, respectively. In FACS analysis, a synergistic induction of ICAM-1 was detectable when cells were co-infected with the two recombinant baculoviruses. These findings clearly demonstrate that IE2p86 and pp71 are crucial regulatory factors for HCMV-induced ICAM-1 upregulation. INTRODUCTIONHuman cytomegalovirus (HCMV), a b-herpesvirus, is a ubiquitous human pathogen with a prevalence of infection in the adult population of 50-90 %. Although HCMV rarely causes severe disease on primary infection in immunocompetent people, it represents a highly pathogenic agent for immunosuppressed patients such as transplant recipients, as well as newborns, and establishes lifelong persistence in the infected host (Alford & Britt, 1990). So far, the exact site of latency is still uncertain; monocytes, haematopoietic precursor cells and endothelial cells have been discussed as potential reservoirs for latent HCMV (Fish et al., 1995;Sindre et al., 1996).Recent evidence suggests that HCMV not only causes acute infection in transplanted patients but may also be associated with extensive transplant vasculopathy, ultimately leading to chronic allograft rejection (Grattan et al., 1989;Skowronski et al., 1993;Koskinen et al., 1993;Lautenschlager et al., 1997a;Schnitzler et al., 1997;Borchers et al., 1999). One of the molecular mechanisms underlying the acceleration of vascular disease processes by HCMV infection may be enhancement of transplant immunogenicity. Several studies indicate that this involves the HCMV-induced upregulation of cellular adhesion molecules, which augments the adherance and infiltration of inflammatory cells that are capable of promoting vascular disease (Einsele et al., 1994;Lemstrom et al., 1995;Steinhoff et al., 1995;Koskinen et al., 1996;Yilmaz et al., 1996; Martelius et al., 1998;Waldman et al., 1998;Lautenschlager et al., 1999;The et al., 2001). For instance, cell culture experiments have demonstrated that...

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Synergistic induction of intercellular adhesion molecule-1 by the human cytomegalovirus transactivators IE2p86 and pp71 is mediated via an Sp1-binding site

Kronschnabl¹,

Stamminger²

2003

Journal of General Virology

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“…Therefore, EC-tropism seems to be determined by the ability to spread from cell to cell rather than to infect EC. In this respect, cell-to-cell spread during propagation of the infection in cell culture, and transfer of virus and virus products from infected EC to uninfected leukocytes, appear to be similar events, both requiring microfusion of the cell membranes of the infected and the contiguous uninfected cell and thus, enabling virus transmission (Gerna et al, 2000). This process may be mediated by an-as-yet unidentified virus gene product directly or indirectly intervening in the cell fusion process.…”

mentioning

confidence: 99%

“…During propagation, a large DNA fragment within a region referred to as ULb9 was lost in strain AD169 and to a slightly lesser extent, in strain Towne (Cha et al, 1996), while another AD169 viral variant, the UK variant, has shown to lack an additional 929 bp in genes UL42 and UL43 (Dargan et al, 1997;Mocarski et al, 1997). Recently, it has been reported that during propagation all laboratory strains have lost some basic biological properties consistently shared by recent clinical HCMV isolates, such as the ability to grow in endothelial cells (EC-tropism) and to be transferred to peripheral blood leukocytes (leukocytetropism or leukotropism), either polymorphonuclear leukocytes (PMNL) or monocytes (Revello et al, 1998;Gerna et al, 2000Gerna et al, , 2002a. It has also been demonstrated that clinical isolates recovered from different body sites lost both properties following a high number of passages in fibroblasts (Revello et al, 2001).…”

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confidence: 99%

Rescue of human cytomegalovirus strain AD169 tropism for both leukocytes and human endothelial cells

Gerna

Percivalle

Sarasini

et al. 2003

Journal of General Virology

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Endothelial cell-tropism-and leukocyte-(polymorphonuclear-and monocyte-) tropism (leukotropism) are two important biological properties shared by all recent clinical isolates of human cytomegalovirus (HCMV). These properties are lost during extensive propagation of HCMV isolates in human fibroblasts, as shown by reference laboratory-adapted strains AD169 and Towne. Here we show that strain AD169 may reacquire both properties in vitro, endothelial (both venous and arterial) cell-tropism preceding leukotropism (predominantly involving monocytes). Restriction fragment length polymorphism analysis and sequencing performed on the original virus inoculum from human fibroblasts and serial passages on endothelial cells confirmed virus identity. Thus, fundamental biological properties may be lost and reacquired in vitro according to the cell culture system employed. The lack of a 15 kb DNA fragment in the strain AD169 genome does not prevent the rescue of these biological functions, thus indicating that they are likely to be encoded by viral genes located elsewhere.

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“…4 This is due to the biologic characteristics of pp65, which is synthesized in excess by HCMV-infected endothelial cells before transfer to leukocytes. 5,6 (3) In the cohort examined in our study, 2 altered viral clearance was observed in 10 of 44 of the patients treated. Three patients (1 in the DNAemia and 2 in the antigenemia arm) had an initial paradoxical rise in antigenemia, which was dissociated from a prompt DNAemia decrease.…”

mentioning

confidence: 99%

Response: Human cytomegalovirus DNAemia and preemptive treatment of HCMV infection in children receiving hematopoietic stem-cell transplantation

Gerna¹,

Lilleri²,

Locatelli³

2008

Blood

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Human Cytomegalovirus Replicates Abortively in Polymorphonuclear Leukocytes after Transfer from Infected Endothelial Cells via Transient Microfusion Events

Cited by 148 publications

References 38 publications

Synergistic induction of intercellular adhesion molecule-1 by the human cytomegalovirus transactivators IE2p86 and pp71 is mediated via an Sp1-binding site

Synergistic induction of intercellular adhesion molecule-1 by the human cytomegalovirus transactivators IE2p86 and pp71 is mediated via an Sp1-binding site

Rescue of human cytomegalovirus strain AD169 tropism for both leukocytes and human endothelial cells

Response: Human cytomegalovirus DNAemia and preemptive treatment of HCMV infection in children receiving hematopoietic stem-cell transplantation

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