Umeå University's logo

umu.sePublikasjoner
Endre søk
RefereraExporteraLink to record
Permanent link

Direct link
Referera
Referensformat
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Annet format
Fler format
Språk
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Annet språk
Fler språk
Utmatningsformat
  • html
  • text
  • asciidoc
  • rtf
Early host cell interactions and antivirals against ocular adenoviruses
Umeå universitet, Medicinska fakulteten, Institutionen för klinisk mikrobiologi, Virologi. (Niklas Arnberg)ORCID-id: 0000-0002-4440-3181
2015 (engelsk)Doktoravhandling, med artikler (Annet vitenskapelig)Alternativ tittel
Tidiga värd cells interaktioner och antiviraler mot okulära adenovirus (svensk)
Abstract [en]

Viruses are common causative agents of ocular infection among humans. Epidemic keratoconjuntivitis (EKC) is a severe and contagious ocular disease with reported outbreaks worldwide. It is estimated that this disease affects 20-40 million individuals every year, which leads to huge socioeconomic costs for the affected countries. EKC is characterized by keratitis and conjunctivitis but is also associated with pain, edema, lacrimation, and decreased vision that can prolong for months after the infection and in rare cases years. This disease is caused by human adenoviruses (HAdVs), which belong to the family of Adenoviridae. Currently, there is no available treatment against EKC.

EKC is mainly caused by HAdV-8, HAdV-19, HAdV-37, HAdV-53, HAdV-54, and HAdV-56, which belong to species D HAdVs. HAdV-8, HAdV-19 and HAdV-37 have previously been shown to use sialic acid (SA)-containing glycans as cellular receptors to bind to and infect human corneal epithelial (HCE) cells. To characterize the receptor in more detail, we performed a glycan array, which included SA-containing glycans. A branched hexasaccharide terminating with SA in each arm was identified as a candidate receptor. This glycan corresponds to the glycan motif found on a ganglioside, GD1a. By performing a series of biological and biochemical experiments we confirmed the function of the GD1a glycan as a cellular receptor for EKC-causing HAdVs. However, the glycan used as a receptor was linked to plasma membrane protein(s) through O-glycosidic bonds, rather than to a lipid (as in the ganglioside). X-ray crystallography analysis showed that the two terminal SA:s interacted with two of the three previously identified SA-binding sites on the knob domain of the HAdV-37 capsid protein known as the fiber.

Based on the structural features of the GD1a:HAdV-37 knob interaction, we assumed that a three-armed molecule with each arm terminating with SA would be an efficient inhibitor. Such molecules were designed, synthesized and found to efficiently prevent HAdV-37 binding to and infection of corneal cells. These results indicate that trisialic acids-containing compounds may be used for treatment of EKC.

After binding to its primary receptor, most HAdVs have been shown to interact with αVβ3 and αVβ5 integrins to enter human cells. This interaction occurs through the RGD (arginine-alanine-aspartic acid) motif in the capsid protein known as the penton base. However, it was not clear if corneal epithelial cells express αVβ3 and αVβ5 integrins. Thus, to better understand additional early steps of infection by EKC-causing HAdVs, we performed binding and infection competition experiments using human corneal epithelial cells and siRNA, integrin specific antibodies, peptides and RGD-containing ligands indicating that α3, αV, β1 affected HAdV-37 infection of but not binding to HCE cells. We could also see that HAdV-37 co-localize with α3 and αV at after entry into HCE cells. In situ histochemistry confirmed that the expression of α3 and αV in human corneal tissue. Overall, our results suggest that αV and α3 integrins are important for HAdV-37 infection of corneal cells.

Altogether, these results provide further insight into the biology of HAdVs and open up for development of novel antiviral drugs.

sted, utgiver, år, opplag, sider
Umeå: Umeå University , 2015. , s. 90
Serie
Umeå University medical dissertations, ISSN 0346-6612 ; 1697
Emneord [en]
Adenovirus, Virus host interactions, Antivirals, Sialic acid, Integrins, Epidemic keratoconjuntivitis
HSV kategori
Identifikatorer
URN: urn:nbn:se:umu:diva-99907ISBN: 978-91-7601-211-6 (tryckt)OAI: oai:DiVA.org:umu-99907DiVA, id: diva2:788590
Disputas
2015-03-13, Hörsal Betula, 6M, Norrlands universitetssjukhus, Umeå, 09:00 (engelsk)
Opponent
Veileder
Tilgjengelig fra: 2015-02-20 Laget: 2015-02-16 Sist oppdatert: 2018-06-07bibliografisk kontrollert
Delarbeid
1. The GD1a glycan is a cellular receptor for adenoviruses causing epidemic keratoconjunctivitis (Letter)
Åpne denne publikasjonen i ny fane eller vindu >>The GD1a glycan is a cellular receptor for adenoviruses causing epidemic keratoconjunctivitis (Letter)
Vise andre…
2011 (engelsk)Inngår i: Nature Medicine, ISSN 1078-8956, E-ISSN 1546-170X, Vol. 17, nr 1, s. 105-109Artikkel i tidsskrift (Fagfellevurdert) Published
Abstract [en]

Adenovirus type 37 (Ad37) is a leading cause of epidemic keratoconjunctivitis (EKC), a severe and highly contagious ocular disease. Whereas most other adenoviruses infect cells by engaging CD46 or the coxsackie and adenovirus receptor (CAR), Ad37 binds previously unknown sialic acid-containing cell surface molecules. By glycan array screening, we show here that the receptor-recognizing knob domain of the Ad37 fiber protein specifically binds a branched hexasaccharide that is present in the GD1a ganglioside and that features two terminal sialic acids. Soluble GD1a glycan and GD1a-binding antibodies efficiently prevented Ad37 virions from binding and infecting corneal cells. Unexpectedly, the receptor is constituted by one or more glycoproteins containing the GD1a glycan motif rather than the ganglioside itself, as shown by binding, infection and flow cytometry experiments. Molecular modeling, nuclear magnetic resonance and X-ray crystallography reveal that the two terminal sialic acids dock into two of three previously established sialic acid-binding sites in the trimeric Ad37 knob. Surface plasmon resonance analysis shows that the knob-GD1a glycan interaction has high affinity. Our findings therefore form a basis for the design and development of sialic acid-containing antiviral drugs for topical treatment of EKC.

HSV kategori
Identifikatorer
urn:nbn:se:umu:diva-38769 (URN)10.1038/nm.2267 (DOI)21151139 (PubMedID)2-s2.0-78651242668 (Scopus ID)
Tilgjengelig fra: 2011-01-03 Laget: 2010-12-28 Sist oppdatert: 2023-04-24bibliografisk kontrollert
2. A Potent Trivalent Sialic Acid Inhibitor of Adenovirus Type 37 Infection of Human Corneal Cells
Åpne denne publikasjonen i ny fane eller vindu >>A Potent Trivalent Sialic Acid Inhibitor of Adenovirus Type 37 Infection of Human Corneal Cells
Vise andre…
2011 (engelsk)Inngår i: Angewandte Chemie International Edition, ISSN 1433-7851, E-ISSN 1521-3773, Vol. 50, nr 29, s. 6519-6521Artikkel i tidsskrift (Fagfellevurdert) Published
sted, utgiver, år, opplag, sider
Wiley, 2011
Emneord
adenoviruses, antiviral agents, crystal-structure elucidation, sialic acids, surface plasmon resonance
HSV kategori
Identifikatorer
urn:nbn:se:umu:diva-45043 (URN)10.1002/anie.201101559 (DOI)21648036 (PubMedID)2-s2.0-79959995269 (Scopus ID)
Tilgjengelig fra: 2011-06-22 Laget: 2011-06-20 Sist oppdatert: 2023-03-24bibliografisk kontrollert
3. Triazole linker-based trivalent sialic acid inhibitors of adenovirus type 37 infection of human corneal epithelial cells
Åpne denne publikasjonen i ny fane eller vindu >>Triazole linker-based trivalent sialic acid inhibitors of adenovirus type 37 infection of human corneal epithelial cells
Vise andre…
2015 (engelsk)Inngår i: Organic and biomolecular chemistry, ISSN 1477-0520, E-ISSN 1477-0539, Vol. 13, nr 35, s. 9194-9205Artikkel i tidsskrift (Fagfellevurdert) Published
Abstract [en]

Adenovirus type 37 (Ad37) is one of the principal agents responsible for epidemic keratoconjunctivitis (EKC), a severe ocular infection that remains without any available treatment. Recently, a trivalent sialic acid derivative (ME0322, Angew. Chem. Int. Ed., 2011, 50, 6519) was shown to function as a highly potent inhibitor of Ad37, efficiently preventing the attachment of the virion to the host cells and subsequent infection. Here, new trivalent sialic acid derivatives were designed, synthesized and their inhibitory properties against Ad37 infection of the human corneal epithelial cells were investigated. In comparison to ME0322, the best compound (17a) was found to be over three orders of magnitude more potent in a cell-attachment assay (IC50 = 1.4 nM) and about 140 times more potent in a cell-infection assay (IC50 = 2.9nM). X-ray crystallographic analysis demonstrated a trivalent binding mode of all compounds to the Ad37 fiber knob. For the most potent compound ophthalmic toxicity in rabbits was investigated and it was concluded that repeated eye administration did not cause any adverse effects.

HSV kategori
Identifikatorer
urn:nbn:se:umu:diva-100014 (URN)10.1039/C5OB01025J (DOI)000360115100007 ()2-s2.0-84940403803 (Scopus ID)
Tilgjengelig fra: 2015-02-18 Laget: 2015-02-18 Sist oppdatert: 2023-03-24bibliografisk kontrollert
4. Involvement of corneal integrins during infection of human adenovirus type 37
Åpne denne publikasjonen i ny fane eller vindu >>Involvement of corneal integrins during infection of human adenovirus type 37
Vise andre…
(engelsk)Manuskript (preprint) (Annet vitenskapelig)
HSV kategori
Identifikatorer
urn:nbn:se:umu:diva-100015 (URN)
Tilgjengelig fra: 2015-02-18 Laget: 2015-02-18 Sist oppdatert: 2018-06-07bibliografisk kontrollert

Open Access i DiVA

fulltext(7209 kB)600 nedlastinger
Filinformasjon
Fil FULLTEXT02.pdfFilstørrelse 7209 kBChecksum SHA-512
f6654bea7541921233fc6f79a4bf48650a62fc6731b64484df257fb365dcbca2bc2c3789bf144be1b148c07e29590b430235d011e3125ea313c16ad1581b7a0f
Type fulltextMimetype application/pdf
spikblad(179 kB)76 nedlastinger
Filinformasjon
Fil SPIKBLAD01.pdfFilstørrelse 179 kBChecksum SHA-512
9a73544113423b700f73bc39787206620cbc34e071a6e52e7170b7ad51bc965a3409eab82633322b85cb92a3015ad67548573fc5858d0e6d129ce636d2625faf
Type spikbladMimetype application/pdf

Person

Storm, Rickard

Søk i DiVA

Av forfatter/redaktør
Storm, Rickard
Av organisasjonen

Søk utenfor DiVA

GoogleGoogle Scholar
Totalt: 600 nedlastinger
Antall nedlastinger er summen av alle nedlastinger av alle fulltekster. Det kan for eksempel være tidligere versjoner som er ikke lenger tilgjengelige

isbn
urn-nbn

Altmetric

isbn
urn-nbn
Totalt: 900 treff
RefereraExporteraLink to record
Permanent link

Direct link
Referera
Referensformat
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Annet format
Fler format
Språk
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Annet språk
Fler språk
Utmatningsformat
  • html
  • text
  • asciidoc
  • rtf