Protein species of the parvovirus minute virus of mice strain MVMp: involvement of phosphorylated VP-2 subtypes in viral morphogenesis

Santarén, Juán F.; Ramírez, Juan Camilo; Almendral, José M.

doi:10.1128/jvi.67.9.5126-5138.1993

Cited by 23 publications

(6 citation statements)

References 80 publications

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“…The adenovirus E4orf6 protein was stained by the anti-E4orf6 (1807) rabbit polyclonal antibody. Monoclonal and polyclonal antibodies raised in rabbit and mouse against MVM structural proteins specifically recognizing assembled capsids (Kaufmann et al, 2007), disassembled VP subunits (Gil-Ranedo et al, 2015), and mature DNA-filled viral particles (Maroto et al, 2004), as well as the major MVM non-structural NS1 protein (Santaré n et al, 1993;Faisst et al 1995;Yeung et al, 1991) have been previously described. The mouse anti-b-III-tubulin (Promega), and rabbit anti-b actin (Thermofisher) polyclonal antibodies were used as loading controls.…”

Section: Antibodiesmentioning

confidence: 99%

Viral targeting of glioblastoma stem cells with patient-specific genetic and post-translational p53 deregulations

2021

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Section: Antibodiesmentioning

confidence: 99%

Viral targeting of glioblastoma stem cells with patient-specific genetic and post-translational p53 deregulations

2021

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“…Antibodies. The MVM antibodies used in this work have been previously described: the ␣-VP rabbit polyclonal antiserum was raised in rabbit against denatured VP2 and used for the general localization of the VP1 and VP2 proteins independently of the configuration (65); the B7 mouse monoclonal antibody (B7-MAb), which recognizes an epitope configured by an intact empty capsid and DNA-filled virus localized at the 3-fold axes (58,59) but fails to react with isolated VP subunits or trimers (28,66); and the ␣-MVM antibody raised in rabbit against the native capsid and used to recognize mainly conformational epitopes (67,89).…”

Section: Methodsmentioning

confidence: 99%

Antiangiogenic Vascular Endothelial Growth Factor-Blocking Peptides Displayed on the Capsid of an Infectious Oncolytic Parvovirus: Assembly and Immune Interactions

Grueso

Sánchez-Martı́nez

Calvo-López

et al. 2019

J Virol

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As many tumor cells synthetize vascular endothelial growth factors (VEGF) that promote neo-vascularization and metastasis, frontline cancer therapies often administer anti-VEGF (α-VEGF) antibodies. To target the oncolytic parvovirus minute virus of mice (MVM) to the tumor vasculature, we studied the functional tolerance, evasion of neutralization, and induction of α-VEGF antibodies of chimeric viruses in which the footprint of a neutralizing monoclonal antibody within the 3-fold capsid spike was replaced by VEGF-blocking peptides: P6L (PQPRPL) and A7R (ATWLPPR). Both peptides allowed viral genome replication and nuclear translocation of chimeric capsid subunits. MVM-P6L efficiently propagated in culture, exposing the heterologous peptide on the capsid surface, and evaded neutralization by the anti-spike monoclonal antibody. In contrast, MVM-A7R yielded low infectious titers and was poorly recognized by an α-A7R monoclonal antibody. MVM-A7R showed a deficient assembly pattern, suggesting that A7R impaired a transitional configuration that the subunits must undergo in the 3-fold axis to close up the capsid shell. The MVM-A7R chimeric virus consistently evolved in culture into a mutant carrying the P6Q amino acid substitution within the A7R sequence, which restored normal capsid assembly and infectivity. Consistent with this finding, anti-native VEGF antibodies were induced in mice by a single injection of MVM-A7R empty capsids, but not by MVM-A7R virions. This fundamental study provides insights to endow an infectious parvovirus with immune antineovascularization and evasion capacities by replacing an antibody footprint in the capsid 3-fold axis with VEGF-blocking peptides, and it also illustrates the evolutionary capacity of single-stranded DNA (ssDNA) viruses to overcome engineered capsid structural restrictions.IMPORTANCETargeting the VEGF signaling required for neovascularization by vaccination with chimeric capsids of oncolytic viruses may boost therapy for solid tumors. VEGF-blocking peptides (VEbp) engineered in the capsid 3-fold axis endowed the infectious parvovirus MVM with the ability to induce α-VEGF antibodies without adjuvant and to evade neutralization by MVM-specific antibodies. However, these properties may be compromised by structural restraints that the capsid imposes on the peptide configuration and by misassembly caused by the heterologous peptides. Significantly, chimeric MVM-VEbp resolved the structural restrictions by selecting mutations within the engineered peptides that restored efficient capsid assembly. These data show the promise of antineovascularization vaccines using chimeric VEbp-icosahedral capsids of oncolytic viruses but also raise safety concerns regarding the genetic stability of manipulated infectious parvoviruses in cancer and gene therapies.

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“…Viral structural proteins were detected with a polyclonal hyperimmune rabbit anti-MVM capsid antiserum that stains denatured structural MVM proteins (41) and also neutralizes infectivity (43). The antiserum was enriched in immunoglobulin G by affinity purification through a protein A-Sepharose column and used at 1/100 dilution in PBS-gelatin.…”

Section: Methodsmentioning

confidence: 99%

Parvovirus minute virus of mice strain i multiplication and pathogenesis in the newborn mouse brain are restricted to proliferative areas and to migratory cerebellar young neurons

Ramírez

Fairén

Almendral

1996

J Virol

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Newborn BALB/c mice intranasally inoculated at birth with a lethal dose of the immunosuppressive strain of the parvovirus minute virus of mice (MVMi) developed motor disabilities and intention tremors with a high incidence by the day 6 postinfection (dpi). These neurological syndromes paralleled the synthesis of virus intermediate DNA replicative forms and yield of infectious particles in the brain, with kinetics that peaked by this time. The preferred virus replicative sites in the brain were established early in the infection (2 dpi) and at the onset of clinical symptoms (6 dpi) and were compared with major regions of cellular proliferative activity found after intraperitoneal injection of bromodeoxyuridine 24 h before encephalons were subjected to immunohistochemistry detection. At 2 dpi, viral capsid antigen was located in the laterodorsal thalamic and the pontine nuclei but not in the extensive proliferative regions of the mouse brain at this postnatal day. At 6 dpi, however, the neurotropism of the MVMi was highlighted by its ability to target the subventricular zone of the ventricles, the subependymal zone of the olfactory bulb, and the dentate gyrus of the hippocampus, which are the three main germinal centers of the cerebrum in mouse postbirth neurogenesis. Unexpectedly, in the cerebellum, the MVMi capsid antigen was confined exclusively to cells that have undergone mitosis and have migrated to the internal granular layer (IGL) and not to the proliferative external granular layer (EGL), which was stained with antiproliferative cell nuclear antigen antibody and is the main target in other parvovirus infections. This result implies temporal or differentiation coupling between MVMi cycle and neuroblast morphogenesis, since proliferative granules of the EGL should primarily be infected but must migrate in a virus carrier state into the IGL in order to express the capsid proteins. During migration, many cells undergo destruction, accounting for the marked hypocellularity specifically found in the IGL and the irregular alignment of Purkinje cell bodies, both consistent histopathological hallmarks of animals developing cerebellar symptoms. We conclude that MVMi impairs postmitotic neuronal migration occurring in the first postnatal week, when, through the natural respiratory route of infection, the virus titer peaks in the encephalon. The results illustrate the intimate connection between MVMi neuropathogenesis and mouse brain morphogenetic stage, underscoring the potential of parvoviruses as markers of host developmental programs.

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Protein species of the parvovirus minute virus of mice strain MVMp: involvement of phosphorylated VP-2 subtypes in viral morphogenesis

Cited by 23 publications

References 80 publications

Viral targeting of glioblastoma stem cells with patient-specific genetic and post-translational p53 deregulations

Viral targeting of glioblastoma stem cells with patient-specific genetic and post-translational p53 deregulations

Antiangiogenic Vascular Endothelial Growth Factor-Blocking Peptides Displayed on the Capsid of an Infectious Oncolytic Parvovirus: Assembly and Immune Interactions

Parvovirus minute virus of mice strain i multiplication and pathogenesis in the newborn mouse brain are restricted to proliferative areas and to migratory cerebellar young neurons

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