Abstract
BackgroundThere are over 100 known human adenovirus (HAdV) types, which are able to cause a broad variety of different self-limiting but also lethal diseases especially in immunocompromised patients. Only limited information about the pathogenesis and biology of the majority of these virus types is available. In the present study, we performed a systematic screen for coxsackievirus and adenovirus receptor (CAR)-usage of a large spectrum of HAdV types.MethodsTo study receptor usage we utilized a recombinant HAdV library containing HAdV genomes tagged with a luciferase and GFP encoding transgene. We infected CHO-CAR cells stably expressing the CAR receptor and control cells lacking the CAR receptor with tagged viruses (HAdV3, 14, 16, 50, 10, 24, 27, 37 and 69) and measured luciferase expression levels 26 and for some viruses (AdV10, − 24 and − 27) 52 h post-infection. As positive control, we applied human adenovirus type 5 (HAdV5) known to use the CAR receptor for cell entry. For viruses replication studies on genome level we applied digital PCR.ResultsInfection of CHO-CAR and CHO-K1 cells at various virus particle numbers per cell (vpc) revealed that HAdV10, 24, and 27 showed similar or decreased luciferase expression levels in the presence of CAR. In contrast, HAdV3, 14, 16, 50, 37 and 69 resulted in increased luciferase expression levels in our initial screening experiments. CAR usage of HAdV3, 14, 50, and 69 was not studied before, and therefore we experimentally confirmed CAR usage for these HAdV as novel viruses utilizing CAR as a receptor. To rule out that replication of HAdV in transduced CHO cells is responsible for increased transduction rates we performed replication assays on virus genome level, which revealed that there is no HAdV replication.ConclusionIn the present study, we screened a HAdV library and identified novel human HAdV using the CAR receptor. To our knowledge, this is the first description of CAR usage for HAdV 3, 14, 50, and 69.
Highlights
In the clinic, human adenoviruses (HAdV) gained increasing importance
Note that for human adenovirus type 5 (HAdV5) it is well established that this virus utilizes coxsackievirus and adenovirus receptor (CAR) for cell entry, and it was applied as a positive control in the present study
We explored Chinese hamster ovarian (CHO)-CAR cells stably expressing the human coxsackievirus and adenovirus receptor and CHO-K1 CAR-negative control cells, which were cultured in DMEM medium (PAN-Biotech GmbH, Aidenbach, Germany) with 10% FCS (PAN-Biotech GmbH, Aidenbach, Germany) and 1% Penicillin/Streptomycin (PAN-Biotech GmbH, Aidenbach, Germany) using 5% CO2 at 37 °C
Summary
Human adenoviruses (HAdV) gained increasing importance. They cause different clinical symptoms with a wide range of diseases such as conjunctivitis, gastroenteritis, pneumonia, and myocarditis. Note that for HAdV5 it is well established that this virus utilizes CAR for cell entry, and it was applied as a positive control in the present study. After infection with respective total virus particle numbers per cell (vpc), all luciferase measurements were performed 26 h post-infection (Fig. 1a).
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