Unlike most adenovirus (Ad) serotypes, the species B Ads do not use the coxsackie-adenovirus receptor as an attachment receptor. The species B attachment receptor(s) has not yet been identified and is also poorly characterized. Species B Ads can be further divided into species B1 and B2 Ads, and these display different organ tropisms, suggesting a difference in receptor usage. We have studied the receptor interactions of the species B1 serotypes 3p and 7p and the species B2 serotypes 11p and 35 and characterized the properties of the species B receptor(s). Reciprocal blocking experiments using unlabeled Ad11p or Ad3p virions to block the binding to A549 cells of (35)S-labeled 3p, 7p, 11p, and 35 showed that only Ad11p virions efficiently blocked the binding of all the species B Ads studied (> or =70%). Thus, there is apparently a common species B Ad receptor (sBAR). However, Ad3p virions only partially (< or =30%) blocked the binding of Ad11p and Ad35 to A549 cells. Binding experiments after trypsin treatment of the cells confirmed that the species B2 serotypes address at least two different receptors on A549 and J82 cells, since sBAR is trypsin sensitive but the species B2 Ad receptor (sB2AR) is not. Both receptors are proteins or glycoproteins, since binding of all species B serotypes was abolished after proteinase K or subtilisin treatment of A549 or J82 cells. Furthermore, binding of the species B serotypes to sBAR was abolished with EDTA and restored with Ca(2+), whereas the binding of Ad11p and Ad35 to SB2AR was independent of divalent cations.

Adenovirus 7 (Ad7) is the adenovirus species that most frequently has been associated with severe illness. Seven distinct genome types of adenovirus 7, Ad7p, Ad7a, Ad7b, Ad7c, Ad7d, Ad7e, and Ad7f, can be identified by using restriction endonucleases BamHI and SmaI. We analyzed the distribution of the different Ad7 genome types among 314 isolates from patients and healthy shedders. The Ad7b and Ad7c genome types accounted for 90% of the isolates from patients and appeared to be mutually exclusive. A shift from Ad7c to Ad7b genome types occurred in 1969 in Europe and in 1975 in Australia. During the last decade, Ad7b genome types predominated in Australia, Europe, and North America. Ad7c was detected in South Africa, Ad7d was detected in China, Ad7e was detected in Brazil, and Ad7f was detected in Australia. The Ad7p and Ad7a genome types dominated among isolates obtained from healthy shedders and appeared scattered through the years and the geographical areas. The prevalence of Ad7 infections is high in Japan as judged by the herd immunity. However, the low percentage (2%) of Adi isolates among all adenovirus isolates chiefly from patients, coupled with 30 to 50% antibody prevalence, argues for a high proportion of inapparent infections and, hence, Ad7 strain(s) of low pathogenicity.

Comparative analysis of the genome organization of human adenovirus 11, a member of the human adenovirus species B, and the commonly used human adenovirus 5 vector, a member of species C Adenovirus type 11 (Ad11), a member of the human adenovirus species B (HAdV-B), has a tropism for the urinary tract. The genome of Ad11 was found to comprise 34 794 bp and is 1141 bp shorter than the Ad5 genome of species HAdV-C. The G+C content of the Ad11 genome is 48?9 %, whereas that of Ad5 is 55?2 %. Ad11 and Ad5 share 57 % nucleotide identity and possess the same four early regions, but the E3 region of Ad11 could not be divided into E3A and E3B. The late genes of Ad11 and Ad5 are organized into six and five regions, respectively. Thirty-eight putative ORFs were identified in the Ad11 genome. The ORFs in the late regions, the E2B region and IVa2 show high amino acid identity between Ad11 and Ad5, whereas the ORFs in E1, E2A, E3 and E4, protein IX and the fibre protein show low amino acid identity. The highest and lowest identities were noted in the pre-terminal protein and fibre proteins: 85 % and 24?6 %, respectively. The E3 20?3K and 20?6K ORFs and the L6 agnoprotein were present in the Ad11 genome only, whereas the E3 11?6K cell death protein was identified only in Ad5. All ORFs but the E3 10?3K and L4 pVIII protein vary not only in composition but also in size. Ad11 may have a higher vector capacity than Ad5, since it has a shorter genome and a shorter fibre. Furthermore, in the E3 region, two additional ORFs can be deleted to give extra capacity for foreign DNA. INTRODUCTIONHuman adenoviruses constitute a large group within the family Adenoviridae, with 51 serotypes identified so far. These serotypes are divided into six species, designated A-F, based on DNA homology (Green et al., 1979; Wadell et al., 1980;Wadell, 1984). The nine human adenovirus members of species B have been classified further into two subspecies, B:1 and B:2. Adenovirus 3 (Ad3), Ad7, Ad16, Ad21 and Ad50 belong to subspecies B:1 and cause respiratory infection, whereas Ad11, Ad34 and Ad35 of subspecies B:2 are associated with persistent infections of the urinary tract and kidney. Ad11 was first isolated from a stool sample of a patient with poliomyelitis and has subsequently been isolated from the urine of patients with haemorrhagic cystitis, as well as from pregnant women. Ad11 can also be found in healthy children and adults. Ad11, Ad34 and Ad35 are strongly over-represented among isolates from immunosuppressed patients and bone marrow transplant recipients.Ad11 targets an unknown cellular receptor, which is more widely distributed than the usual coxsackie-adenovirus receptor (CAR) and is highly expressed on the surface of cells from various organs. Furthermore, relatively few persons have been exposed to Ad11 and, consequently, seroprevalence is low (D'Ambrosio et al., 1982). Previously, we demonstrated that Ad11 binds strongly to and replicates in kidney cells (HEK 293 cells), an endothelial cell line, and committed haematopoietic cell lines. ...

Adenovirus infections are widespread in society and are occasionally associated with severe, but rarely with life-threatening, disease in otherwise healthy individuals. In contrast, adenovirus infections present a real threat to immunocompromised individuals and can result in disseminated and fatal disease. The number of patients undergoing immunosuppressive therapy for solid organ or hematopoietic stem cell transplantation is steadily increasing, as is the number of AIDS patients, and this makes the problem of adenovirus infections even more urgent to solve. There is no formally approved treatment of adenovirus infections today, and existing antiviral agents evaluated for their antiadenoviral effect give inconsistent results. We have developed a whole cell-based assay for high-throughput screening of potential antiadenoviral compounds. The assay is unique in that it is based on a replication-competent adenovirus type 11p green fluorescent protein (GFP)-expressing vector (RCAd11pGFP). This allows measurement of fluorescence changes as a direct result of RCAd11pGFP genome expression. Using this assay, we have screened 9,800 commercially available small organic compounds. Initially, we observed approximately 400 compounds that inhibited adenovirus expression in vitro by >80%, but only 24 were later confirmed as dose-dependent inhibitors of adenovirus. One compound in particular, 2-{[2-(benzoylamino)benzoyl]amino}-benzoic acid, turned out to be a potent inhibitor of adenovirus replication.

Adenoviruses of six subgenera, namely, adenovirus 31 (Ad31) (subgenus A), Ad3, Ad7, Ad11p, Ad11a, and Ad35 (subgenus B), Ad5v and Ad5p (subgenus C), Ad37 (subgenus D), Ad4 (subgenus E), and Ad41 (subgenus F), were studied. The relative binding properties of different adenoviruses to 293 (human kidney embryonic cells) and A549 (human lung carcinoma cells) cells were compared by flow cytometry. All analyzed adenoviruses bound to cells in a dose-dependent manner. The binding capacity showed that Ad11p, Ad35 (subgenus B:2) with kidney tropism, and Ad4 (subgenus E), which can cause adenopharyngoconjunctivitis, bound strongly to both A549 and 293 cells. The other members of subgenus B and Ad37 of subgenus D manifested an intermediate binding capacity. The analyzed adenoviruses of subgenera A, C, and F manifested a low affinity. Adenoviruses of subgenera B:2 and E manifested high binding affinity to preparations of cell membranes from the epithelial cell lines. Reciprocal competition experiments using Ad11p and Ad4 demonstrated that the two viruses did not block each other. Antibodies against alphavbeta3 and alphavbeta5 reduced the binding of Ad5v virions and slightly impaired the binding of Ad4 but did not affect Ad11p binding to the A549 cell surface. Recombinant fiber proteins of Ad11p and Ad35 reciprocally blocked the binding of both viruses to the epithelial cells but they could not block Ad4. The hexon protein expression of Ad11p and Ad4 was 100 times more efficient than that of the Ad5 vector (pFG140), whereas the infectivity of Ad11p and Ad4 was 40- to 200-fold that of the commonly used Ad5v vector. Taken together, our findings demonstrate that Ad11p and Ad4 bind different receptor molecules and that the fibers of these two viruses provide the predominant high degree of binding, which obviously is a requirement for subsequent internalization and efficacious expression.