Monoclonal Antibodies against the Adeno-Associated Virus Type 2 (AAV-2) Capsid: Epitope Mapping and Identification of Capsid Domains Involved in AAV-2–Cell Interaction and Neutralization of AAV-2 Infection

Wobus, Christiane E.; Hügle-Dörr, Barbara; Girod, Anne; Petersen, Gabriele; Hallek, Michael; Kleinschmidt, Jürgen A.

doi:10.1128/jvi.74.19.9281-9293.2000

Cited by 251 publications

(338 citation statements)

References 51 publications

Supporting

Mentioning

322

Contrasting

Order By: Relevance

“…The surface expo- sure of these residues was also predicted based on a model by Arbetman et al (13). The surface-localized residues occupy analogous regions to those mapped as antigenic footprints on other AAV capsids, for example, AAV2 and AAV8 (97,100,101,114), and thus likely dictate the antigenic differences reported between the two viruses when their transduction efficiencies were compared in the presence of intravenous immunoglobulin (13). Internal capsid volumes are comparable between AAV2, AAV4, and AAV5 and do not support an increased packaging capacity for AAV5 compared to other AAVs.…”

Section: Fig 2 Aav Capsid Surfaces (A Cmentioning

confidence: 99%

“…The AAV capsid surface features that are unique to AAV5-shorter 3-fold protrusions, extended VR-VII, and smaller HI loop-contain amino acid residues or are proximate to capsid regions reported to play essential roles in the AAV life cycle. For example, residues within VR-IV, VR-V, and VR-VIII which make up the 3-fold protrusions have been reported to control glycan receptor attachment (VR-V and VR-VIII for AAV2 and VR-V for AAV9) (90)(91)(92)(93)122), transduction (VR-IV, VR-V, and VR-VIII in AAV2, VR-VIII in AAV8, and VR-IV, VR-V, and VR-VIII in AAV9) (94)(95)(96)(97)(98)(99) and antigenic phenotypes (VR-IV, VR-V, and VR-VIII in AAV2 and VR-VIII in AAV8 (97,100,101). These regions have similar roles in AAV5.…”

Section: Fig 2 Aav Capsid Surfaces (A Cmentioning

confidence: 99%

See 1 more Smart Citation

Structural Insights into Adeno-Associated Virus Serotype 5

Govindasamy

DiMattia

Gurda

et al. 2013

J Virol

View full text Add to dashboard Cite

The adeno-associated viruses (AAVs) display differential cell binding, transduction, and antigenic characteristics specified by their capsid viral protein (VP) composition. Toward structure-function annotation, the crystal structure of AAV5, one of the most sequence diverse AAV serotypes, was determined to 3.45-Å resolution. The AAV5 VP and capsid conserve topological features previously described for other AAVs but uniquely differ in the surface-exposed HI loop between ␤H and ␤I of the core ␤-barrel motif and have pronounced conformational differences in two of the AAV surface variable regions (VRs), VR-IV and VR-VII. The HI loop is structurally conserved in other AAVs despite amino acid differences but is smaller in AAV5 due to an amino acid deletion. This HI loop is adjacent to VR-VII, which is largest in AAV5. The VR-IV, which forms the larger outermost finger-like loop contributing to the protrusions surrounding the icosahedral 3-fold axes of the AAVs, is shorter in AAV5, creating a smoother capsid surface topology. The HI loop plays a role in AAV capsid assembly and genome packaging, and VR-IV and VR-VII are associated with transduction and antigenic differences, respectively, between the AAVs. A comparison of interior capsid surface charge and volume of AAV5 to AAV2 and AAV4 showed a higher propensity of acidic residues but similar volumes, consistent with comparable DNA packaging capacities. This structure provided a three-dimensional (3D) template for functional annotation of the AAV5 capsid with respect to regions that confer assembly efficiency, dictate cellular transduction phenotypes, and control antigenicity. R ecombinant adeno-associated viruses (rAAVs) are promising viral vectors for gene delivery applications (1, 2). These viruses belong to the single-stranded DNA (ssDNA)-packaging Parvoviridae and genus Dependovirus. Hundreds of AAV genotypes have been sequenced from several mammalian species, and to date 12 serotypes (AAV1 to AAV12) have been defined for the human and nonhuman primate isolates (3-15). The 12 serotypes are classified into eight genetic groups, clades A to F and clonal isolates AAV4 and AAV5, based on antigenic reactivity and sequence similarity (15). The groups are represented by AAV1 to AAV9, with AAV1 and AAV6 belonging to the same clade A because of their high sequence similarity and antigenic cross-reactivity. The representative members share ϳ55 to 99% sequence identity, with AAV4 and AAV5 being the most divergent from each other and from the other members.The linear ssDNA AAV genome, ϳ4.7 kb in length, contains two genes: cap, which encodes the capsid viral proteins (VPs; VP1, ϳ87 kDa; VP2, ϳ73 kDa; VP3, ϳ62 kDa) and rep, which encodes the replication proteins necessary for genome replication and genome packaging. Inverted terminal repeats (ITRs) at the end of the AAV genome, 145 bp in length, are the only essential active sequence required to function as (i) the origin for DNA replication, (ii) the packaging signal, and (iii) integration sites (16)(17)(18)(19). Recombina...

show abstract

Section: Fig 2 Aav Capsid Surfaces (A Cmentioning

confidence: 99%

Section: Fig 2 Aav Capsid Surfaces (A Cmentioning

confidence: 99%

Structural Insights into Adeno-Associated Virus Serotype 5

Govindasamy

DiMattia

Gurda

et al. 2013

J Virol

View full text Add to dashboard Cite

show abstract

“…Peptide-scanning and phage display experiments have been utilized to identify numerous dominant antigenic regions of the AAV2 capsid, i.e., epitopes that several neutralizing antibodies apparently bind (Moskalenko et al, 2000;Wobus et al, 2000). To prevent antibody binding and subsequent neutralization, the surface properties of these, and likely other, regions of the AAV capsid must be altered.…”

Section: Introductionmentioning

confidence: 99%

PEG conjugation moderately protects adeno-associated viral vectors against antibody neutralization

Lee

Maheshri

Kaspar³

et al. 2005

Biotechnol. Bioeng.

132

114

View full text Add to dashboard Cite

AAV gene therapy vectors have significant clinical promise, but serum neutralization poses a challenge that must be overcome. We have examined the potential of conjugating the AAV surface with activated polyethylene glycol chains to protect the vector from neutralizing antibodies. Two key parameters were investigated: the polymer chain size and the PEG:lysine conjugation ratio. Transduction data revealed that the vector is fully infectious until a critical PEG conjugation reaction ratio was exceeded, and this critical level was found to vary with polymer chain size. At this key conjugation ratio, however, particles were moderately protected from serum neutralization, 2.3-fold over unmodified vector, demonstrating that there is a small window of PEGylation for which particles are still fully infective and benefit from antibody protection. TEM results and structural analysis indicate that the drop of infectivity as the PEG concentration is increased beyond the critical conjugation ratio may be due to a combination of steric interference with viral regions necessary for infection as well as reaction at important lysine residues. However, this first study analyzing the potential of PEG to protect AAV from serum neutralization shows that the approach has promise, which can be further enhanced if the locations of PEG attachment can be more finely controlled. ß

show abstract

“…44 The epitopes for three AAV2-specific murine neutralizing monoclonal antibodies were mapped, and found to differ from those defined above by human polyclonal NABs. 72 The consensus between the above two studies is that no single linear epitope was responsible for AAV2 neutralization, even for a single monoclonal antibody. This suggests that NAB might recognize conformational epitopes, making this strategy more difficult to bring to fruition.…”

Section: Identification Of Neutralizing Aav Capsid Epitopesmentioning

confidence: 99%