Human bocavirus 1 infection of CACO-2 cell line cultures

Ghietto, Lucía María; D'Angelo, Ana Paola Toigo; Viale, Franco Agustin; Adamo, María Pilar

doi:10.1016/j.virol.2017.07.034

Cited by 9 publications

(8 citation statements)

References 43 publications

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“…The finding that T84 cells, an immortalized intestinal cell line, are readily transduced with all five BoV vectors is notable, as Caco-2 cells, another intestinal cell line, do not support productive infection 50 . Moreover, HAEs or CuFi-8 cells, the only other two cell culture systems that are currently available for ex vivo BoV research, are considerably more difficult to obtain, culture, and polarize.…”

Section: Resultsmentioning

confidence: 99%

Novel Chimeric Gene Therapy Vectors Based on Adeno-Associated Virus and Four Different Mammalian Bocaviruses

Fakhiri

Schneider

Puschhof

et al. 2019

Molecular Therapy - Methods & Clinical Development

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Parvoviruses are highly attractive templates for the engineering of safe, efficient, and specific gene therapy vectors, as best exemplified by adeno-associated virus (AAV). Another candidate that currently garners increasing attention is human bocavirus 1 (HBoV1). Notably, HBoV1 capsids can cross-package recombinant (r)AAV2 genomes, yielding rAAV2/HBoV1 chimeras that specifically transduce polarized human airway epithelia (pHAEs). Here, we largely expanded the repertoire of rAAV/BoV chimeras, by assembling packaging plasmids encoding the capsid genes of four additional primate bocaviruses, HBoV2–4 and GBoV (Gorilla BoV). Capsid protein expression and efficient rAAV cross-packaging were validated by immunoblotting and qPCR, respectively. Interestingly, not only HBoV1 but also HBoV4 and GBoV transduced pHAEs as well as primary human lung organoids. Flow cytometry analysis of pHAEs revealed distinct cellular specificities between the BoV isolates, with HBoV1 targeting ciliated, club, and KRT5+ basal cells, whereas HBoV4 showed a preference for KRT5+ basal cells. Surprisingly, primary human hepatocytes, skeletal muscle cells, and T cells were also highly amenable to rAAV/BoV transduction. Finally, we adapted our pipeline for AAV capsid gene shuffling to all five BoV isolates. Collectively, our chimeric rAAV/BoV vectors and bocaviral capsid library represent valuable new resources to dissect BoV biology and to breed unique gene therapy vectors.

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Section: Resultsmentioning

confidence: 99%

Novel Chimeric Gene Therapy Vectors Based on Adeno-Associated Virus and Four Different Mammalian Bocaviruses

Fakhiri

Schneider

Puschhof

et al. 2019

Molecular Therapy - Methods & Clinical Development

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“…Nucleic acids were extracted from 200 µl of RS and serum samples using guanidinium buffer and silica, as described previously [21,22] . Extracts were stored at −20 °C for later use in subsequent end-point PCR assays for detection of HBoV1, targeting a 354 bp sequence of the NP1 gene as performed previously [3,22,23] . PCR products were visualized in 8.5 % polyacrylamide gels stained with silver solution (0.11 M AgNO 3 ) [24] .…”

Section: Nucleic Acid Extraction and Hbov1 Detectionmentioning

confidence: 99%

Diagnosis and clinical significance of Human bocavirus 1 in children hospitalized for lower acute respiratory infection: molecular detection in respiratory secretions and serum

Salbetti

Boggio

Abbiatti

et al. 2022

Journal of Medical Microbiology

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Introduction. Human bocavirus 1 (HBoV1) infection occurs with viral genome presence in respiratory secretions (RS) and serum, and therefore both samples can be used for diagnosis. Gap statement. The diagnostic sensitivity of HBoV1 DNA detection in serum and the duration of DNAaemia in severe clinical cases have not been elucidated. Aim. To determine HBoV1 DNA in serum and RS of paediatric patients hospitalized for lower acute respiratory infection (LARI) and to analyse the clinical–epidemiological features of positive cases. Methodology. This was a prospective, transverse study. Physicians selected the clinical situations and obtained paired clinical samples (RS and serum) that were tested by PCR/qPCR for HBoV1. Positive cases were analysed considering time of specimen collection, co-detection, clinical manifestations and viral load; statistical significant level was set at α=0.05. Results. HBoV1 was detected in 98 of 402 cases included (24 %); 18/98 (18 %) patients had the virus detectable in serum and 91/98 (93 %) in RS (P<0.001). Positivity rates were not significantly different in patients with RS and serum collected within or beyond 24 h of admission. Single HBoV1 infection was identified in 39/98 patients (40 %), three patients had HBoV1 in both clinical samples (3/39, 8 %) and 32 (32/39, 82 %) only in RS, 22 of them (69 %) with both clinical samples within 24 h of admission. Cough (P=0.001) and rhinitis (P=0.003) were significantly frequent among them and most patients were diagnosed with bronchiolitis (22/39, 56 %) and pneumonia (9/39, 23 %), which was more frequent compared to cases with co-infection (P=0.04). No significant differences were identified among patients with high, medium or low viral load of HBoV1 regarding rate of positivity in both clinical samples, the time of collection of RS and serum, co-detection, first episode of LARI, clinical manifestations, comorbidity or requirement for assisted ventilation. Intensive care unit (ICU) patients had a significantly higher frequency of detection (P<0.001) and co-detection (P=0.001) compared to patients on standard care. Conclusions. HBoV1 is prevalent among infant patients hospitalized for LARI and including it in the standard testing can add to the aetiological diagnosis in these cases, especially for patients admitted to the ICU. HBoV1 detection in serum did not contribute significantly to the diagnosis as compared to detection in respiratory secretions.

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“…Recently, Ghietto and coworkers proposed that CaCo‐2 cells, a cell line derived from a colorectal tumor, could be used to study parts of the HBoV replication cycle, as these cells could be infected with HBoV and shed the virus 7. Without doubts, this is a major progress, as CaCo‐2 cells are easily available and do not require extensive and expensive cell culturing efforts.…”

mentioning

confidence: 99%