Biology subjects

van den Doel, P. B.

Publications and source records attributed to van den Doel, P. B..

4 recordsLinked to original sources

bioRxiv2021

Human airway cells prevent SARS-CoV-2 multibasic cleavage site cell culture adaptation

Virus propagation methods generally use transformed cell lines to grow viruses from clinical specimens, which may force viruses to rapidly adapt to cell culture conditions, a process facilitated by high viral mutation rates. Upon propagation in VeroE6 cells, SARS-CoV-2 may mutate or delete the multibasic cleavage site (MBCS) in the spike protein that facilitates serine protease-mediated entry into human airway cells. We report that propagating SARS-CoV-2 on the human airway cell line Calu-3 - that expresses serine proteases - prevents MBCS mutations. Similar results were obtained using a human airway organoid-based culture system for SARS-CoV-2 propagation. Thus, in-depth knowledge on the biology of a virus can be used to establish methods to prevent cell culture adaptation.

microbiology↗

bioRxiv2020

Elephant endotheliotropic herpesvirus is omnipresent in elephants in European zoos and an Asian elephant range country

Elephant endotheliotropic herpesviruses (EEHVs) are a group of evolutionary divergent herpesviruses that may cause acute, often lethal, hemorrhagic disease (EEHV-HD) in young elephants. Although EEHV was first discovered over 20 years ago, its prevalence in different elephant populations is still largely unknown, partially due to the lack of readily available, sensitive serological assays. In order to improve diagnostic tools for the detection of EEHV infections and to obtain insight in its spread among elephants, we developed novel ELISAs focusing on EEHV1A gB and gH/gL as antigens. Performance of the ELISAs was assessed using sera taken from 41 European zoo elephants and 69 semi-captive elephants from Laos, one of the Asian elephant range countries. Sera from all (sub)adult animals tested ([≥]5 years of age) showed high reactivity with both gB and gH/gL, whereas reactivity towards the antigens was generally lower for sera of juvenile animals (1 > 5 years). Only one (juvenile) animal, which was sampled directly after succumbing to EEHV-HD, was found to be seronegative for EEHV. The two other EEHV-HD cases tested showed low antibody levels, suggesting that all three cases died upon a primary EEHV infection. Direct comparison with another EEHV-specific ELISA previously used in two large serosurveys, showed that EEHV prevalence was underestimated before, likely due to aberrant folding of the antigen used. In conclusion, our study suggests that essentially all (semi-)captive (sub)adult elephants in European zoos and in Laos carry EEHV, and that young elephants with low antibody levels are at risk of dying from EEHV-HD. ImportanceOver the last 30 years, nearly 20% of all Asian elephants born in Western zoos succumbed to acute hemorrhagic disease caused by elephant endotheliotropic herpesvirus (EEHV-HD). Yet, the prevalence of EEHV in captive and wild elephant populations is still largely unknown, mainly due to the lack of readily available, sensitive serological assays. For this study two highly sensitive EEHV-specific ELISAs were developed. Using these assays, it was shown that nearly all elephants tested were seropositive for EEHV, with highest antibody levels detected in (sub)adult elephants. In contrast, antibody levels in EEHV-HD cases were very low or non-detectable. Lack of antibodies may thus be a risk factor for developing severe disease. As the novel ELISAs are low-tech in nature, these assays may easily be disseminated to local laboratories in zoos and elephant range countries in order to determine EEHV serostatus of individual animals or complete herds and (wild) populations.

microbiology↗

bioRxiv2020

The SARS-CoV-2 multibasic cleavage site facilitates early serine protease-mediated entry into organoid-derived human airway cells

After the SARS-CoV outbreak in 2003, a second zoonotic coronavirus named SARS-CoV-2, emerged late 2019 in China and rapidly caused the COVID-19 pandemic leading to a public health crisis of an unprecedented scale. Despite the fact that SARS-CoV-2 uses the same receptor as SARS-CoV, transmission and pathogenesis of both viruses seem to be quite distinct. A remarkable feature of the SARS-CoV-2 spike is the presence of a multibasic cleavage site, which is absent in the SARS-CoV spike. The viral spike protein not only attaches to the entry receptor, but also mediates fusion after cleavage by host proteases. Here, we report that the SARS-CoV-2 spike multibasic cleavage site increases infectivity on differentiated organoid-derived human airway cells. Compared with SARS-CoV, SARS-CoV-2 entered faster into the lung cell line Calu-3, and more frequently formed syncytial cells in differentiated organoid-derived human airway cells. Moreover, the multibasic cleavage site increased entry speed and plasma membrane serine protease usage relative to endosomal entry using cathepsins. Blocking serine protease activity using the clinically approved drug camostat mesylate effectively inhibited SARS-CoV-2 entry and replication in differentiated organoid-derived human airway cells. Our findings provide novel information on how SARS-CoV-2 enters relevant airway cells and highlight serine proteases as an attractive antiviral target. Significance StatementHighly pathogenic coronaviruses have spilled from animals to humans three times in the past two decades. Late 2019, SARS-CoV-2 emerged in China and was declared a pandemic by March 2020. The other two highly pathogenic coronaviruses, SARS-CoV and MERS-CoV, emerged in 2002 and 2012, respectively, but did not attain sustained human-to-human transmission. Given the high diversity of coronaviruses in animals, urbanization and increased air travel, future coronavirus pandemics are likely to occur intermittently. Identifying which factors determine pandemic potential and pathogenicity are therefore of key importance to global health. Additionally, there is an urgent need to rapidly translate fundamental knowledge to the clinic, a process that is expedited through the use of relevant cell culture systems.

microbiology↗

bioRxiv2020

Susceptibility of rabbits to SARS-CoV-2

Transmission of severe acute respiratory coronavirus-2 (SARS-CoV-2) between livestock and humans is a potential public health concern. We demonstrate the susceptibility of rabbits to SARS-CoV-2, which excrete infectious virus from the nose and throat upon experimental inoculation. Therefore, investigations on the presence of SARS-CoV-2 in farmed rabbits should be considered.

microbiology↗