Utilization of Vero Cells for Primary and Chronic BK Virus Infection

Acott, Philip D.; O’Regan, Patrick A.; Lee, S.H.; Crocker, John F. S.

doi:10.1016/j.transproceed.2006.10.163

Cited by 10 publications

(25 citation statements)

References 6 publications

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“…Infections of Vero cells were then performed. Vero cells were chosen for the infection since they are able to sustain productive replication of BKV (Acott et al, 2006) and have been widely used for basic research purposes, providing important data regarding virus binding to its receptor, the infectivity profile of different BKV strains and the effect of specific drugs on viral replication (Acott et al, 2006; Sinibaldi et al, 1987, 1990). Cells infected with the same starting number of genomic copies for each viral strain were allowed to grow for 59 days with occasional subculturing every 6 to 8 days.…”

Section: Resultsmentioning

confidence: 99%

“…Moreover, the onset of rearrangements within the NCCR may also depend on the cell line used for the in vitro propagation of the virus. Acott et al showed that changes within the NCCR of several BKV isolates propagated in Vero cells started to appear at around 50 days P.I., suggesting that genomic modifications may become frequent only after a prolonged infection course (Acott et al, 2006). …”

Section: Resultsmentioning

confidence: 99%

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Polymorphisms of the BK virus subtypes and their influence on viral in vitro growth efficiency

Tremolada

Delbue

Larocca³

et al. 2010

Virus Research

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SUMMARYThe major capsid protein, VP1, of the human Polyomavirus BK (BKV) is structurally divided into five outer loops, referred to as BC, DE, EF, GH, and HI. The BC loop includes a short region, named the BKV subtyping region, spanning nucleotides 1744-1812 and characterized by non-synonymous nucleotide polymorphisms that have been used to classify different strains of BKV into four subtypes. The aim of this study was to determine if the nucleotide changes clustered within the BKV subtyping region may influence the in vitro growth efficiency of the virus. We therefore infected the African Green Monkey kidney cell line Vero with four different viral strains (named BKV I, II, III, and IV) that contained the nucleotide sequences of the BKV subtypes within the same genomic background. Infected cells were followed for 59 days and viral replication was assessed at different time points by Quantitative Real Time PCR (Q-PCR). BKV I, II, and IV were successfully propagated over time in Vero cells, whereas BKV III viral loads progressively decreased during the infection course, demonstrating that the non-synonymous nucleotide polymorphisms of subtype III confer a strong disadvantage for viral replication. Since subtype III differs from all the other subtypes at position 68 of the VP1, where Leu is replaced by Gln, we created viral strains bearing Gln at this position together with the polymorphisms of subtypes I, II, IV and tested their growth in Vero cells. Our results demonstrate that this amino acid substitution does not lower the replication efficiency of subtypes I, II, and IV. In conclusion, this study provides further insights to the importance of the BC loop of BKV in the virus life cycle. In addition, given the effect of the amino acid substitutions of the four BKV subtypes on infectious spread of the virus, our results suggest the need to investigate their potential association with BKV related complications.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Polymorphisms of the BK virus subtypes and their influence on viral in vitro growth efficiency

Tremolada

Delbue

Larocca³

et al. 2010

Virus Research

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“…All previous BKPyV anti-viral experiments have been performed in human or monkey kidney cells or lung fibroblasts (12, 35, 36). Based on our observation that BKPyV can also infect salivary gland cells in vitro and is associated with pathology in vivo in humans (28, 29, 45, 46), three drugs with demonstrated anti BKPyV activity in kidney cells, CPRO, CDV and LEF were assessed for BKPyV inhibition in salivary gland cells in vitro.…”

Section: Discussionmentioning

confidence: 99%

“…Assays to test anti-BKPyV drugs have previously been performed in kidney or lung cells (12, 35, 36). Noting the importance of BKPyV in HIV-SGD and the detection of efficient BKPyV replication in salivary gland cells in vitro (28, 29), it is imperative that an in vitro drug screen against BKPyV replication in human salivary gland cells be tested to identify effective viral targets for drug treatment for salivary gland derived BKPyV.…”

Section: Introductionmentioning

confidence: 99%

Effect of Leflunomide, Cidofovir and Ciprofloxacin on replication of BKPyV in a salivary gland in vitro culture system

2015

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BK polyomavirus (BKPyV) is a known kidney tropic virus that has been detected at high levels in HIV-associated salivary gland disease (HIV-SGD), one of the most important AIDS associated oral lesions. BKPyV has been detected in HIV-SGD patient saliva and replicates in salivary gland cells in vitro. BKPyV antivirals are currently in wide use to guard against BKPyV mediated organ rejection in kidney transplant recipients. The goal of this study was to investigate the inhibitory effects of three such antiviral agents, ciprofloxacin, cidofovir, and leflunomide in BKPyV infected salivary gland cells. Human salivary gland cells, and Vero cells, were infected with BKPyV, treated with antiviral drugs and assessed for BKPyV gene expression and viral replication for up to 5 days post infection. The kinetics of BKPyV replication were different in salivary gland cells compared to kidney cells. Ciprofloxacin and cidofovir had minimal effect on metabolic activity and host cell DNA replication, however, cell toxicity was detected at the protein level with leflunomide treatment. Ciprofloxacin decreased BKV T Ag and VP1 mRNA expression by at least 50% in both cell types, and decreased T Ag protein expression at days 3 and 4 post infection. A 2.5 – 4 log decrease in intracellular DNA replication and a 2 – 3 log decrease in progeny release were detected with ciprofloxacin treatment. Cidofovir and leflunomide also inhibited BKPyV gene expression and DNA replication. The three drugs diminished progeny release by 30–90% and 2 – 6 fold decreases in infectious virus were detected post drug treatment by fluorescence focus assay. Additionally, three clinical BKPyV isolates were assessed for their responses to these agents in vitro. Cidofovir and leflunomide, but not ciprofloxacin treatment resulted in statistically significant inhibition of BKPyV progeny release from salivary gland cells infected with HIV-SGD BKPyV isolates. All three drugs decreased progeny release from cells infected with a transplant derived viral isolate. In conclusion, treatment of human salivary gland cells with each of the three drugs produced modest decreases in BKPyV genome replication. These data highlight the need for continued studies to discover more effective and less toxic drugs that inhibit BKPyV replication in salivary gland cells.

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“…On the one hand, they result in longer transplant retention, at the same time, however, they lead to the activation of opportunistic pathogens in the recipient. One of them is the BK virus that carries a risk of developing nephropathy and a risk of graft loss [5,6]. Hence, there is the need to seek an effective antiviral therapy that will prevent the development of viraemia/ viruria, thus avoiding subsequent complications.…”

Section: Introductionmentioning

confidence: 99%

Antiviral activity of ciprofloxacin on BK virus in an in vitro culture – pilot study

Pasternak¹,

Rajtar²,

Polz‐Dacewicz³

2017

J Pre Clin Clin Res.

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Introduction. Reactivation of BKV infection caused by decrease of immunity associated with organ transplantation can lead to nephropathy and organ loss. Antiviral drugs such as ganciclovir or foscarnet did not display any activity against BK virus. A source of hope can be found in chemotherapeutics usually used in bacterial infections that inhibit the replication of viral genetic material in cells. One of the aforementioned chemotherapeutics is ciprofloxacin. The aim of the study is to evaluate the antiviral activity of ciprofloxacin against BK virus in vitro culture. Materials and method. Initially, the effects of ciprofloxacin on Vero cell viability were assessed with MTT. After determining the 3 largest non-cytotoxic concentrations of antibiotics, their effect on viral replication in cells was determined. 24-h cells culture in DMEM medium supplemented with 10% serum after 2h incubation with a 6.3 x 10 5 copies/ml infection virus were used to evaluate the effects of different concentrations of ciprofloxacin on the number of BKV copies. Quantification of BKV replication in the culture was performed by real-time PCR. Results. By means of the MTT test, the 3 largest non-cytotoxic ciprofloxacin concentrations were determined as follows: 31.25; 62.5; 125 μg/ml. Cells treated with ciprofloxacin at a concentration of 125 μg/ml display the greatest decrease in BKV replication that equaly 1.8 -3 log/5.8 -9.2 Ct, compared to the control of the virus without drugs. A decline in the number of copies of the virus was observed during the lower concentration of the antibiotic (62.5, 31.25 µg/ml) by 1.3 -2 log/4 -6.4 Ct and 0 -0.8 log/0 -2, 4 Ct, respectively. Conclusions. Ciprofloxacin exhibits antiviral activity against the BK virus by inhibiting its replication in the in vitro culture in Vero cell line.

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Utilization of Vero Cells for Primary and Chronic BK Virus Infection

Cited by 10 publications

References 6 publications

Polymorphisms of the BK virus subtypes and their influence on viral in vitro growth efficiency

Polymorphisms of the BK virus subtypes and their influence on viral in vitro growth efficiency

Effect of Leflunomide, Cidofovir and Ciprofloxacin on replication of BKPyV in a salivary gland in vitro culture system

Antiviral activity of ciprofloxacin on BK virus in an in vitro culture – pilot study

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