Identification of novel human adenovirus candidates using the coxsackievirus and adenovirus receptor for cell entry

Abstract

Background There 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. Methods To 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. Results Infection 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. Conclusion In 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.

Adenovirus Core Proteins: Structure and Function

“…Adenoviruses (AdVs) are non-enveloped icosahedral viruses with a double-stranded DNA genome [ 1 ]. Since its discovery in 1953 [ 2 , 3 ], more than 120 species-specific adenoviral serotypes have been identified in humans, mammals, birds, fish and reptiles [ 4 , 5 ]. Though human adenoviruses are not generally associated with causing severe disease in immunocompetent humans, they may cause severe infections in immunocompromised people [ 6 , 7 , 8 ].…”

Variation in the affinity of three representative avian adenoviruses for the cellular coxsackievirus and adenovirus receptor

“…CAR is a transmembrane protein that mediates cell–cell adhesion and contains two glycosylated extracellular Ig-like domains, D1 and D2, followed by a transmembrane domain [ 6 ] and an intracellular domain (ICD) [ 7 ]. CAR is a high-affinity primary receptor and coreceptor for human adenovirus (HAdV) groups A and C–F, as well as canine and gorilla adenovirus [ 8 – 10 ].…”

Properties of Adenovirus Vectors with Increased Affinity to DSG2 and the Potential Benefits of Oncolytic Approaches and Gene Therapy

“…Based on the distinguishing features of JO4 viruses in cancer cell lines with different DSG2 levels, we hypothesized that not only DSG2 binding but also interaction with other molecules or the lack thereof played a crucial role in the life cycle of JO4 viruses. In addition to DSG2, CD46 has proven to be a minor receptor for Ad3 infection, and CAR was also hypothesized as such [33,34,47]. To decode the mechanism behind the unexpected weakening effect of the JO4 mutation on the virus, we further explored these viruses in stable CD46-and CAR-expressing CHO cell lines.…”