A rapid, simple, and accurate plaque assay for human respiratory syncytial virus (HRSV)

Kim, Kyung Sook; Kim, Ah Ra; Piao, Ying; Lee, Ju Hie; Quan, Fu‐Shi

doi:10.1016/j.jim.2017.03.020

Cited by 8 publications

(6 citation statements)

References 15 publications

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“…Amongst the several ways of viral inhibition analysis, the plaque assay, immunofluorescence microscopy, and qPCR were considered as standard tools [ 13 , 21 , 49 ]. The anti-RSV effect was therefore confirmed in the plaque assay ( Figure 5 ), immunofluorescence imaging ( Figure 6 ), and qRT-PCR.…”

Section: Discussionmentioning

confidence: 99%

Anti-RSV Peptide-Loaded Liposomes for the Inhibition of Respiratory Syncytial Virus

et al. 2018

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Although respiratory syncytial virus (RSV) is one of the leading causes of acute respiratory tract infection in infants and adults, effective treatment options remain limited. To circumvent this issue, there is a novel approach, namely, the development of multifunctional liposomes for the delivery of anti RSV-peptides. While most of the peptides that are used for loading with the particulate delivery systems are the penetrating peptides, an alternative approach is the development of liposome-peptide systems, which are loaded with an RSV fusion peptide (RF-482), which has been designed to inhibit the RSV fusion and block infection. The results of this work have revealed that the liposomes themselves can serve as potential RSV inhibitors, whilst the anti-RSV-peptide with liposomes can significantly increase the RSV inhibition when compared with the anti-RSV peptide alone.

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Section: Discussionmentioning

confidence: 99%

Anti-RSV Peptide-Loaded Liposomes for the Inhibition of Respiratory Syncytial Virus

et al. 2018

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“…The plaque reduction neutralization assay was performed with some modifications ( 19 ). Briefly, HEp-2 cells were seeded onto a 96-well plate at a density of 3.5 × 10 5 cells/mL in DMEM containing antibiotic-antimycotic solution (1%) and 10% FBS and grown in a CO 2 incubator until the cells reached >95% confluency.…”

Section: Methodsmentioning

confidence: 99%

“…The plaque reduction neutralization assay was performed with some modifications (19 The percentage functionality loss of palivizumab at a digestion point was determined with respect to the unincubated sample in the ex vivo study, whereas it was determined with respect to feed in the in vivo study. Percentage functionality was determined for each of the three experimental replicates separately and these values were used for statistical analyses.…”

Section: Determination Of Neutralization Titer Of Samples Against Rsvmentioning

confidence: 99%

Partial Degradation of Recombinant Antibody Functional Activity During Infant Gastrointestinal Digestion: Implications for Oral Antibody Supplementation

et al. 2020

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Oral administration of engineered immunoglobulins has the potential to prevent enteric pathogen-induced diarrhea in infants. To prevent infection, these antibodies need to survive functionally intact in the proteolytic environment of the gastrointestinal tract. This research examined both ex vivo and in vivo the functional survival across infant digestion of palivizumab, a model FDA-approved recombinant antibody against respiratory syncytial virus (RSV) F protein. Palivizumab-fortified feed (formula or human milk), infant gastric, and intestinal samples were incubated to simulate in vivo digestion (ex vivo digestion). Palivizumab-fortified human milk was also fed to infants, followed by collection of gastric and intestinal samples (in vivo digestion). Palivizumab was purified from the samples of digestate using protein G spin columns followed by filtration through molecular weight cutoff membranes (30 kDa). Palivizumab functional survival across ex vivo and in vivo digestion was determined via an anti-idiotype ELISA and an RSV plaque reduction neutralization test. Palivizumab concentration and RSV neutralization capacity both decreased when incubated in intestinal samples (ex vivo study). The concentration and neutralization activity of orally-supplemented palivizumab also decreased across infant digestion (in vivo study). These results indicate that if recombinant IgGs were selected for oral supplementation to prevent enteric infections, appropriate dosing would need to account for degradation occurring in the digestive system. Other antibody formats, structural changes, or encapsulation could enhance survival in the infant gastrointestinal tract.

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“…HEp-2 cells were seeded in a 12-well cell culture plate (SPL Life Sciences, Korea) and cultured until confluent. All procedures were followed as previously described (Kim et al ., 2017). Serially diluted RSV stocks were used to infect the cells and overlaid with 1% noble agar and incubated at 37 °C, 5% CO 2 .…”

Section: Methodsmentioning

confidence: 99%

The resistance againstTrichinella spiralisinfection induced by primary infection with respiratory syncytial virus

et al. 2018

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Human infections withTrichinella spiralisand respiratory syncytial virus (RSV) are common, asT. spiralisinfections are re-emerging in various parts of the world and RSV infections remain a threat for infants. Yet, studies investigating the relationship pertaining to the two are severely lacking. In particular, immune response inductionviaRSV andT. spiralisremain largely elusive. Here, we investigated the resistance againstT. spiralisinfection induced upon primary infection with RSV. RSV, notorious for causing severe inflammatory reaction in the lungs, were intranasally infected, followed with aT. spiralisinfection in mice. Our results revealed that primary RSV infection in mice significantly raisedT. spiralis-specific and total IgE, IgG and its subclass antibody responses uponT. spiralischallenge infection (RSV-Ts). Blood eosinophil levels were decreased in RSV-Ts, accompanied with significant increase in both Th1 and Th2 cytokines. Antibodies generated against RSV in RSV-infected mice were found to react withT. spiralisexcretory/secretory antigen, showing several bands determined through immunoblotting. RSV-Ts also had a marked reduction ofT. spiralisworm burden in diaphragm. These results indicate that immune responses induced by RSV infection contribute to resistance against subsequentT. spiralisinfection.

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A rapid, simple, and accurate plaque assay for human respiratory syncytial virus (HRSV)

Cited by 8 publications

References 15 publications

Anti-RSV Peptide-Loaded Liposomes for the Inhibition of Respiratory Syncytial Virus

Anti-RSV Peptide-Loaded Liposomes for the Inhibition of Respiratory Syncytial Virus

Partial Degradation of Recombinant Antibody Functional Activity During Infant Gastrointestinal Digestion: Implications for Oral Antibody Supplementation

The resistance againstTrichinella spiralisinfection induced by primary infection with respiratory syncytial virus

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