Ultrastructural study of human herpesvirus-7 replication in tissue culture

Klußmann, Jens Peter; Krueger, Eugene W.; Sloots, Theo P.; Berneman, Zwi; Arnold, Georg J.; Krueger, G. R. F.

doi:10.1007/s004280050051

Cited by 23 publications

(10 citation statements)

References 33 publications

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Section: Discussionmentioning

confidence: 99%

“…Electron microscopical examinations of infected cells in both Crassostrea gigas and Ostrea edulis suggest that empty capsids accept the virus DNA molecule and that DNA is packaged into a toroidal structure, thereby forming a mature capsid. Empty capsids are also a frequent finding, which may reflect a discrepancy between the production of virus DNA and structural proteins (Klussmann et al 1997).After replication of the virus, the assembled nucleocapsids of herpesviruses acquire envelopes from the inner lamella of the nuclear, cytoplasmic or cell membranes. Two mechanisms have been described for the envelopment process of herpesvirus capsids (Roizman 1982).…”

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confidence: 99%

“…The rarity of virions in the perinuclear space may reflect either a rapid passage of particles from the nucleus to cytoplasm or low-grade virus replication. The second mechanism involves the apparent prolifer-180 9a 9b 10a 10b ation of the nuclear membranes that react with and thereby envelop mature nucleocapsids (Dargan 1986, Nii 1992, Klussmann et al 1997. Convoluted endoplasmic reticulum and perinuclear membranes are prominent in the cytoplasm of virus-infected cells in C. gigas and O. edulis spat.…”

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confidence: 99%

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Concomitant herpes-like virus infections in hatchery-reared larvae and nursery-cultured spat Crassostrea gigas and Ostrea edulis

Renault

Deuff²,

Chollet³

et al. 2000

Dis. Aquat. Org.

104

101

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Concomitant sporadic high mortalities were reported in France in May 1994 among batches of hatchery-reared larval Pacific oysters Crassostrea gigas and European flat oysters Ostrea edulis in 2 hatcheries, and in June and July 1994 among batches of cultured spat of both species in a shellfish nursery. Histological observation showed the presence of cellular abnormalities in moribund animals. Transmission electron microscopy revealed the presence of herpes-like virus particles in infected larvae and spat of both oyster species. This is the first description of a herpes-like virus infection in larval O. edulis. Viruses observed in diseased larvae and spat of both species are similar with respect to ultrastructure and morphogenesis. They were detected simultaneously in C. gigas and O. edulis larvae and spat, indicating possible interspecific transmission. Moreover, these viruses are associated with high mortality rates in both oyster species. An electron microscopic examination revealed hemocytes with condensed chromatin and extensive perinuclear fragmentation of chromatin. These data suggest that herpes-like viruses infecting oysters may induce apoptosis in oyster hemocytes. KEY WORDS: Herpes-like virus · European flat oyster · Ostrea edulis · Pacific oyster · Crassostrea gigas · Oyster mortality · Concomitant infection · Virus replication · Apoptosis Resale or republication not permitted without written consent of the publisherDis Aquat Org 42: [173][174][175][176][177][178][179][180][181][182][183] 2000 viral particles reported by Hine et al. (1992) in New Zealand and the virus-like particles described by Comps & Cochennec (1993) among French cultured European flat oyster spat.We describe for the first time a herpes-like virus infecting European flat oyster larvae and concomitant herpes-like virus infections associated with high mortalities among hatchery-reared larvae and nurserycultured spat of both Crassostrea gigas and Ostrea edulis during the summer of 1994 in France. We performed an ultrastructural comparative study between both virus-like particles found respectively in Pacific oysters and European flat oysters and compared them to viruses belonging to the Herpesviridae family. MATERIALS AND METHODS Specimens.Larval Ostrea edulis and Crassostrea gigas, 6 to 10 d old, 80 to 150 µm in size, were collected in May 1994 from 2 hatcheries located in Bourgneuf Bay (Vendée, France). Nursery-cultured spat of C. gigas and O. edulis, 3 to 5 mo old, were also collected from Bourgneuf Bay in June and July 1994.Light microscopy. A total of 197 European flat oyster spat and 196 Pacific oyster spat were collected for light microscopical examination. Moribund cultured spat of both species, 3 to 5 mo old, were removed from the shell and, sagittally sectioned; then half of each specimen was fixed in Davidson's fixative for light microscopic examination and the other half in Carson's fixative for transmission electron microscopic analysis. After 48 h fixation in Davidson's fluid, samples were dehydrated using an ...

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Concomitant herpes-like virus infections in hatchery-reared larvae and nursery-cultured spat Crassostrea gigas and Ostrea edulis

Renault

Deuff²,

Chollet³

et al. 2000

Dis. Aquat. Org.

104

101

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“…This demonstrates that infective HHV-7 particles were present in the supernatant of HHV-7-infected macrophage cultures at a given time point (7 days) but decreased thereafter. HHV-7 is a T-lymphotropic virus which has been shown to replicate in CD4 ϩ T lymphocytes and in the SupT1 cell line in vitro (3,11). In vivo studies have shown that HHV-7 antigen can be found in a number of tissues, such as lung, skin, and mammary gland (7), and the salivary gland is proposed to be a major site for virus replication (10).…”

Section: Fig 2 Pcr and Rt-pcr Amplification Of Hhv-7mentioning

confidence: 99%

“…ϩ T cells (6,11). So far, only CD4 ϩ T lymphocytes (isolated from PBMC and cord blood mononuclear cells) and SupT1 cells, immature T-cells derived from a non-Hodgkin's lymphoma, have proved sensitive to HHV-7 infection (2,12).…”

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Productive Infection of Primary Macrophages with Human Herpesvirus 7

Zhang

Bolle

Aquaro

et al. 2001

J Virol

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Here we demonstrate replication of human herpesvirus 7 (HHV-7), a T-lymphotropic virus, in macrophages. Productive replication was lost after 2 weeks, but HHV-7 DNA was detected up to 1 month after infection. Thus, macrophages become infected by HHV-7 and might play an important role as a viral reservoir, as has been demonstrated for human immunodeficiency virus type 1.Human herpesvirus 7 (HHV-7) was first isolated in 1990 from peripheral blood mononuclear cells (PBMC), under conditions required to promote T-cell activation, from a healthy individual and later on also from a patient with chronic fatigue syndrome (3, 5). HHV-7 utilizes CD4 as its main receptor to enter the target cell and has also been shown to interfere with human immunodeficiency virus type 1 infection in CD4 ϩ T cells (6, 11). So far, only CD4 ϩ T lymphocytes (isolated from PBMC and cord blood mononuclear cells) and SupT1 cells, immature T-cells derived from a non-Hodgkin's lymphoma, have proved sensitive to HHV-7 infection (2, 12). Lack of productive HHV-7 infection in CD4 ϩ macrophages was observed by quantitative PCR and immunofluorescence assays to detect HHV-7 antigen in vitro (4). It has also been demonstrated that HHV-7 antigen is expressed in macrophages/ monocytes of AIDS-associated Kaposi's sarcoma samples (8). Another, more recent study has shown that in lymph nodes of AIDS patients HHV-7 is mainly present in macrophages (9). This suggests that macrophages can be infected with HHV-7 and may be a site of latency and reactivation of HHV-7 in vivo.In the present study, we evaluated HHV-7 infection of macrophages in vitro. Our results suggest that HHV-7 can infect macrophages and that this infection may persist in these cells for a long time. Macrophages were isolated by density gradient centrifugation from PBMC from healthy blood donors and further enriched, as described in detail elsewhere (1). More than 99% of the cells were CD14 ϩ CD4 ϩ CD3 Ϫ , and no HHV-7 DNA was detected by PCR in these cell cultures. The stock of HHV-7 (strain KHR) was grown in CD8-depleted PBMC, as described previously (13). The CD4 ϩ T-cell line SupT1, the CD4 Ϫ cell line HSB-2, the CD4 ϩ cell line MT-2, and the monocytic CD4ϩ THP-1 cell line were purchased from the American Type Culture Collection (Rockville, Md.).To study the HHV-7 infection in macrophages, the cells were inoculated with virus stock for 4 h and then extensively washed. The SupT1, HSB-2, MT-2, and THP-1 cell lines were also used to determine HHV-7 infection. The mock-infected and HHV-7-infected macrophage cultures were evaluated regularly by light microscopy. PCR, reverse transcription (RT)-PCR, and electron microscopy were also used to detect HHV-7 DNA, mRNA, and HHV-7 particles.DNA was isolated from mock-infected and HHV-7-infected macrophages at different time points after HHV-7 infection. Briefly, the cells were washed extensively with phosphate-buffered saline and collected, and total DNA of the cell pellet was isolated using QIAamp DNA blood Mini Kit (Qiagen, Hilden, Germany) following ...

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