Mutations acquired during cell culture isolation may affect antigenic characterisation of influenza A(H3N2) clade 3C.2a viruses

As elsewhere, few (< 15%) sentinel influenza A(H3N2) clade 3C.2a viruses that dominated in Canada during the 2014/15 season could be antigenically characterised by haemagglutination inhibition (HI) assay. Clade 3C.2a viruses that could be HI-characterised had acquired genetic mutations during in vitro cell culture isolation that modified the potential glycosylation motif found in original patient specimens and the consensus sequence of circulating viruses at amino acid positions 158–160 of the haemagglutinin protein. Caution is warranted in extrapolating antigenic relatedness based on limited HI findings for clade 3C.2a viruses that continue to circulate globally.

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Evaluation of Diallyl Sulfide Analogs for Cytotoxicity, Inhibition of CYP2E1, and Metabolite Profiling Using In Vitro Cell Culture, Human Liver Microsomes and LCMS/MS

10.26226/morressier.57d6b2b8d462b8028d88ee12 ◽

2016 ◽

Author(s):

Mohammad Rahman

Keyword(s):

Cell Culture ◽

Metabolite Profiling ◽

Human Liver ◽

Liver Microsomes ◽

Human Liver Microsomes ◽

Diallyl Sulfide ◽

In Vitro Cell Culture

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Comparison of pCR Rate After Neadjuvant Chemotherapy Guided by Result of in Vitro Cell Culture Drug Sensitivity or Physician's Choice

Case Medical Research ◽

10.31525/ct1-nct04130750 ◽

2019 ◽

Author(s):

Keyword(s):

Cell Culture ◽

Drug Sensitivity ◽

In Vitro Cell Culture

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An In Vitro Cell Culture Model for Pyoverdine-Mediated Virulence

Pathogens ◽

10.3390/pathogens10010009 ◽

2020 ◽

Vol 10 (1) ◽

pp. 9

Author(s):

Donghoon Kang ◽

Natalia V. Kirienko

Keyword(s):

Cell Culture ◽

Virulence Factors ◽

Model Organisms ◽

Cell Culture Model ◽

Chronic Infections ◽

In Vitro Cell Culture ◽

Mitochondrial Homeostasis ◽

Culture Model ◽

Wide Range

Pseudomonas aeruginosa is a multidrug-resistant, opportunistic pathogen that utilizes a wide-range of virulence factors to cause acute, life-threatening infections in immunocompromised patients, especially those in intensive care units. It also causes debilitating chronic infections that shorten lives and worsen the quality of life for cystic fibrosis patients. One of the key virulence factors in P. aeruginosa is the siderophore pyoverdine, which provides the pathogen with iron during infection, regulates the production of secreted toxins, and disrupts host iron and mitochondrial homeostasis. These roles have been characterized in model organisms such as Caenorhabditis elegans and mice. However, an intermediary system, using cell culture to investigate the activity of this siderophore has been absent. In this report, we describe such a system, using murine macrophages treated with pyoverdine. We demonstrate that pyoverdine-rich filtrates from P. aeruginosa exhibit substantial cytotoxicity, and that the inhibition of pyoverdine production (genetic or chemical) is sufficient to mitigate virulence. Furthermore, consistent with previous observations made in C. elegans, pyoverdine translocates into cells and disrupts host mitochondrial homeostasis. Most importantly, we observe a strong correlation between pyoverdine production and virulence in P. aeruginosa clinical isolates, confirming pyoverdine’s value as a promising target for therapeutic intervention. This in vitro cell culture model will allow rapid validation of pyoverdine antivirulents in a simple but physiologically relevant manner.

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A microstrip resonator based sensor for GHz characterization of in vitro cell culture

2018 12th International Conference on Sensing Technology (ICST) ◽

10.1109/icsenst.2018.8603622 ◽

2018 ◽

Cited By ~ 1

Author(s):

F. Kolbl ◽

N. Boulboul ◽

M. Commereuc ◽

E. Bourdel

Keyword(s):

Cell Culture ◽

Microstrip Resonator ◽

In Vitro Cell Culture

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Characterization of an enhanced antigenic change in the pandemic 2009 H1N1 influenza virus haemagglutinin

Journal of General Virology ◽

10.1099/vir.0.061598-0 ◽

2014 ◽

Vol 95 (5) ◽

pp. 1033-1042 ◽

Cited By ~ 5

Author(s):

Blanca García-Barreno ◽

Teresa Delgado ◽

Sonia Benito ◽

Inmaculada Casas ◽

Francisco Pozo ◽

...

Keyword(s):

Influenza Virus ◽

Amino Acid ◽

Influenza A ◽

Point Mutations ◽

Antigenic Site ◽

Single Amino Acid ◽

Antigenic Structure ◽

Human Antibody ◽

2009 H1n1

Murine hybridomas producing neutralizing mAbs specific to the pandemic influenza virus A/California/07/2009 haemagglutinin (HA) were isolated. These antibodies recognized at least two different but overlapping new epitopes that were conserved in the HA of most Spanish pandemic isolates. However, one of these isolates (A/Extremadura/RR6530/2010) lacked reactivity with the mAbs and carried two unique mutations in the HA head (S88Y and K136N) that were required simultaneously to eliminate reactivity with the murine antibodies. This unusual requirement directly illustrates the phenomenon of enhanced antigenic change proposed previously for the accumulation of simultaneous amino acid substitutions at antigenic sites of the influenza A virus HA during virus evolution (Shih et al., Proc Natl Acad Sci USA, 104 , 6283–6288, 2007). The changes found in the A/Extremadura/RR6530/2010 HA were not found in escape mutants selected in vitro with one of the mAbs, which contained instead nearby single amino acid changes in the HA head. Thus, either single or double point mutations may similarly alter epitopes of the new antigenic site identified in this work in the 2009 H1N1 pandemic virus HA. Moreover, this site is relevant for the human antibody response, as shown by competition of mAbs and human post-infection sera for virus binding. The results are discussed in the context of the HA antigenic structure and challenges posed for identification of sequence changes with possible antigenic impact during virus surveillance.

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