Respiratory Syncytial Virus Infection of Human Mononuclear Leukocytes in Vitro and in Vivo

Domurat, Frank M.; Roberts, Norbert J.; Walsh, Edward E.; Dagan, Ron

doi:10.1093/infdis/152.5.895

Cited by 92 publications

(45 citation statements)

References 20 publications

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“…Recently, RS virus has been shown to replicate in vitro in human peripheral blood mononuclear leukocytes (Krilov et al, 1987). In addition, RS virus antigen has been found in circulating mononuclear leukocytes of infants with RS virus infections (Dumorat et al, 1985). The present report describes the infection of the U937 human macrophage cell line by RS virus and the occurrence of virus-enhancing antibodies in human sera.…”

mentioning

confidence: 74%

In vitro Enhancement of Respiratory Syncytial Virus Infection of U937 Cells by Human Sera

Gimenez

Keir

Cash

1989

Journal of General Virology

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SUMMARYHuman sera containing respiratory syncytial (RS) virus-specific antibodies enhance RS virus infection of the U937 macrophage cell line. There was an increase in the number of cells expressing virus antigen when U937 cells were infected with RS virus in the presence of human serum compared to cells infected in the absence of human serum. Human sera enhanced virus yield, as measured by the cell-released infectious virus, by an average of 50-fold compared to virus infection in the absence of human serum. The comparison of the enhancing activities of paired acute and convalescent human sera showed that the titre of enhancing antibody increased in parallel with the titre of RS virus-specific antibody measured by complement fixation and virus neutralization. An RS virus-specific neutralizing monoclonal antibody directed to the virus F protein enhanced virus infection of U937 cells. A non-neutralizing monoclonal antibody directed to the virus nucleoprotein did not enhance virus infection. The possible role of enhancing antibodies in vivo is discussed. INTRODUCTIONRespiratory syncytial (RS) virus is a major cause of acute respiratory infections in young children. The use of a formalin-inactivated RS virus vaccine predisposed children to severe illness when subsequently infected with RS virus (Kim et al., 1969). Following immunization, children developed lower neutralizing antibody titres than those of a comparable age who had natural RS virus infections and had not been immunized (Murphy et al., 1986). Chin et al. (1969) found higher virus shedding and virus isolation rates in vaccinees compared to unvaccinated children when both groups were naturally infected with RS virus. These data led to the speculation that low antibody titres contributed to the severity of the RS virus infections. A possible explanation for these data is antibody-dependent enhancement of virus infection, as proposed by Porterfield (1982) and Halstead (1982), although no experimental data were presented at that time.The in vitro enhancement of virus infection of macrophage cell lines by virus-specific antibodies at sub-neutralizing concentrations has been reported for several viruses (reviewed by Halstead, 1982). The initial stages of virus enhancement require the formation of a virusantibody complex which then attaches, via the Fc portion of the antibody, to the macrophage Fc receptor; this facilitates the entry of the virus as compared to virus infection in the absence of antibody (Gollins & Porterfield, 1984). The sites of replication of RS virus in the respiratory tract are poorly understood and the role of antibody-dependent enhancement in RS virus pathogenesis remains to be determined. Recently, RS virus has been shown to replicate in vitro in human peripheral blood mononuclear leukocytes (Krilov et al., 1987). In addition, RS virus antigen has been found in circulating mononuclear leukocytes of infants with RS virus infections (Dumorat et al., 1985). The present report describes the infection of the U937 human macrophage cell line by...

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confidence: 74%

In vitro Enhancement of Respiratory Syncytial Virus Infection of U937 Cells by Human Sera

Gimenez

Keir

Cash

1989

Journal of General Virology

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“…Detection of BRSV was based on data published on the related HRSV Studies on HRSV showed that in vitro the virus propagates in blood monocytes and alveolar macrophages (Domurat et al 1985;Midulla et al 1989). Viral mRNA was isolated from circulating mononuclear cells of children developing signs of respiratory infection (O'Donnell et al 1989) and HRSV genome was demonstrated by PCR in blood samples (Rohwedder et al 1998).…”

Section: Discussionmentioning

confidence: 99%

Detection of Bovine Respiratory Syncytial Virus in Cell Cultures by nRT-PCR and Use of the Method for Virus Identification in Clinical Samples

Valentová¹,

Kovařčík²,

Pšikal³

2003

Acta Vet. Brno

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One-step RT-PCR and nested PCR were used to detect bovine respiratory syncytial virus (BRSV) in infected cell cultures. Specific 984 bp and 383 bp products, selected from gene encoding the F protein using suitable primers, were amplified and, if appropriate, reamplified. The method was further used for examination of nasal swabs and blood samples collected from animals showing signs of a respiratory disease. Lactating cows and 6-8-month-old bulls from two herds were investigated.The infection by BRSV was confirmed by a) demonstration of specific fragments of BRSV genome in all animals showing signs of an acute disease, and b) indirectly by serological methods in convalescent animals.Viral RNA was detected from nasal swabs and leukocytes of affected animals. The positive PCR product obtained by nested RT-PCR from RNA isolated from leukocytes was sequenced and nucleotide and amino acid homology were 97-100% and 94.5-100% respectively, when compared with sequences from the GenBank.Considering the difficulties associated with the demonstration of BRSV in cell cultures, the amplification techniques provide an effective tool for the identification of the causative agent of bovine respiratory diseases. Nested PCR; BRSV, F protein; diagnosticsBovine respiratory syncytial virus is a pneumovirus of the family Paramyxoviridae causing an acute respiratory disease in cattle. BRSV mostly attacks calves, in which the infection is manifested by pyrexia, anorexia, increased respiratory rate, and dyspnoea. Infections in lactating cows, accompanied by considerable decrease in milk yield by up to 60%, have also been described (Elvander 1996;Pritchard et al. 1997). Morbidity is high and death rate can reach up to 20% (Kovafiãík 1997). BRSV is regarded as the major causative agent of the bovine respiratory syndrome in West European states and the USA. The agent was also demonstrated in the local cattle herds (Pospí‰il et al. 1978; Kovafiãík 1999), but its prevalence in the Czech Republic has not yet been studied.The diagnosis is based on virus demonstration in cell cultures and serological methods for the demonstration of viral antigen and seroconversion. The methods used most frequently for antigen demonstration include the immunofluorescence test and the enzymatic immunosorbent assay (EIA) (Kimman et al. 1986;Krilov et al. 1988;Valarcher et al. 1999). Virus isolation is complicated by a high sensitivity of the agent and the fact that several passages are necessary before CPE develops. Tests on two blood samples collected at three-week intervals are necessary for the demonstration of seroconversion. Moreover, the interpretation of results is impeded by interaction with maternal antibodies present in blood sera of calves. Therefore, the serological methods are used mostly for the demonstration of past infections.

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“…Studies in vitro have shown that Sendal virus inhibits the ability of peripheral blood monocytes to enhance lymphocyte transformation to phytohaemaggtutinin and leads to significant interferon ~ (HulFN-~) production by monocytes, whereas RSV did not affect either of these monocyte functions (Chonmaitree et al, 1981 ;Morahan et al, 1985). Despite this lack of effect on monocytes, RSV has recently been shown to be capable of infecting human peripheral blood monocytes in vitro and RSV antigen has been found in circulating monocytes of infants with RSV disease (Domurat et al,, 1985).…”

Section: Discussionmentioning

confidence: 99%

Respiratory Virus Infection of Peripheral Blood Monocytes: Correlation with Ageing of Cells and Interferon Production in vitro

Krilov

Hendry

Godfrey

et al. 1987

Journal of General Virology

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SUMMARYThe ability of respiratory syncytial virus (RSV) and parainfluenza virus type 3 (PIV3) to replicate in peripheral blood monocytes cultured in vitro for 1, 2, 4 or 7 days prior to infection was investigated. Inoculation of 1-day old monocytes produced at least tenfold less new virus than infection of the older, more macrophage-like cells for both viruses. PIV3 induced extensive syncytium formation, whereas RSV caused a cytopathic effect manifest by increased rounding of the cells with minimal syncytium formation. Supernatants of infected monocytes were assayed for human interferon-c~ (HuIFN-c0 in an attempt to explain the restricted viral replication in the youngest monocytes. In PIV3-infected cells, HuIFN-c~ production was inversely correlated with new virus formation. Monocytes infected after 1 day in culture produced 800 IU/ml of HuIFN-c~; the older cells produced 100 to 200 IU/ml. In contrast, monocytes infected on day l with RSV produced minimal amounts (1.5 IU/ml) of HulFN-c~. Increasing amounts of HulFN-c~ were detected in cells infected with RSV after 2, 4 or 7 days in culture, reaching a maximum of 400 IU/ml on day 7. Further investigation of the apparent restriction of replication in young monocyte cultures may be helpful in understanding the pathogenesis of these respiratory infections.

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Respiratory Syncytial Virus Infection of Human Mononuclear Leukocytes in Vitro and in Vivo

Cited by 92 publications

References 20 publications

In vitro Enhancement of Respiratory Syncytial Virus Infection of U937 Cells by Human Sera

In vitro Enhancement of Respiratory Syncytial Virus Infection of U937 Cells by Human Sera

Detection of Bovine Respiratory Syncytial Virus in Cell Cultures by nRT-PCR and Use of the Method for Virus Identification in Clinical Samples

Respiratory Virus Infection of Peripheral Blood Monocytes: Correlation with Ageing of Cells and Interferon Production in vitro

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