Human transbodies to VP40 inhibit cellular egress of Ebola virus-like particles

Teimoori, Salma; Seesuay, Watee; Jittavisutthikul, Surasak; Chaisri, Urai; Sookrung, Nitat; Densumite, Jaslan; Saelim, Nawannaporn; Chulanetra, Monrat; Maneewatch, Santi; Chaicumpa, Wanpen

doi:10.1016/j.bbrc.2016.09.052

Cited by 19 publications

(12 citation statements)

References 33 publications

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“…More recently, cyclic arginine rich motifs have shown promise as efficient sdAb delivery motifs ( 47 ) though these assemblies required in vitro ligation of the motif to the antibody, which may complicate scale-up. Despite these caveats, one group has pioneered monomeric transbodies to several viruses including influenza A ( 48 ), HCV ( 49 ), and more recently even Ebola ( 50 ) which is encouraging us to pursue the transition of Xintrabodies to Xtransbodies.…”

Section: Discussionmentioning

confidence: 99%

Intracellular Crosslinking of Filoviral Nucleoproteins with Xintrabodies Restricts Viral Packaging

Sherwood

Hayhurst

2017

Front. Immunol.

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Viruses assemble large macromolecular repeat structures that become part of the infectious particles or virions. Ribonucleocapsids (RNCs) of negative strand RNA viruses are a prime example where repetition of nucleoprotein (NP) along the genome creates a core polymeric helical scaffold that accommodates other nucleocapsid proteins including viral polymerase. The RNCs are transported through the cytosol for packaging into virions through association with viral matrix proteins at cell membranes. We hypothesized that RNC would be ideal targets for crosslinkers engineered to promote aberrant protein–protein interactions, thereby blocking their orderly transport and packaging. Previously, we had generated single-domain antibodies (sdAbs) against Filoviruses that have all targeted highly conserved C-terminal regions of NP known to be repetitively exposed along the length of the RNCs of Marburgvirus (MARV) and Ebolavirus (EBOV). Our crosslinker design consisted of dimeric sdAb expressed intracellularly, which we call Xintrabodies (X- for crosslinking). Electron microscopy of purified NP polymers incubated with purified sdAb constructs showed NP aggregation occurred in a genus-specific manner with dimeric and not monomeric sdAb. A virus-like particle (VLP) assay was used for initial evaluation where we found that dimeric sdAb inhibited NP incorporation into VP40-based VLPs whereas monomeric sdAb did not. Inhibition of NP packaging was genus specific. Confocal microscopy revealed dimeric sdAb was diffuse when expressed alone but focused on pools of NP when the two were coexpressed, while monomeric sdAb showed ambivalent partition. Infection of stable Vero cell lines expressing dimeric sdAb specific for either MARV or EBOV NP resulted in smaller plaques and reduced progeny of cognate virus relative to wild-type Vero cells. Though the impact was marginal at later time-points, the collective data suggest that viral replication can be reduced by crosslinking intracellular NP using relatively small amounts of dimeric sdAb to restrict NP packaging. The stoichiometry and ease of application of the approach would likely benefit from transitioning away from intracellular expression of crosslinking sdAb to exogenous delivery of antibody. By retuning sdAb specificity, the approach of crosslinking highly conserved regions of assembly critical proteins may well be applicable to inhibiting replication processes of a broad spectrum of viruses.

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Section: Discussionmentioning

confidence: 99%

Intracellular Crosslinking of Filoviral Nucleoproteins with Xintrabodies Restricts Viral Packaging

Sherwood

Hayhurst

2017

Front. Immunol.

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“…Examples of CPPs are (1) protein transduction domains (PTDs) such as penetratin (PEN; synonym antennapedia homeodomain peptide of Drosophila melanogaster ) [ 105 ], HIV-1 Tat peptide: Tat 49–57 [ 106 , 107 ], transportan (a 27 residue-peptide from galanin neuropeptide and mastoparan or wasp venom toxin) [ 97 ], and VP-22 peptide of structural protein of herpes simplex virus [ 108 ]; (2) amphipathic peptides such as noncytotoxic sweet arrow peptide (SAP) which is a proline-rich motif (VRLPPP) [ 105 ], peptide vector named MPG derived from the fusion sequence of HIV-1 gp41, and a hydrophilic domain of SV40 nuclear localization sequence [ 101 ]; and (3) other CPP type such as nonaarginine (R9) and poly-lysine [ 109 ]. In our laboratory, cell penetrable human scFvs and humanized-camel VHs/VHHs specific to viral proteins and toxins have been prepared by linking the antibody molecules to either penetratin or R9 [ 84 , 85 , 103 , 104 , 110 – 112 ]. These fusion proteins readily entered mammalian cells without causing cytotoxicity and bound to their respective intracellular targets.…”

Section: Therapeutic Antibodiesmentioning

confidence: 99%

Evolution of Therapeutic Antibodies, Influenza Virus Biology, Influenza, and Influenza Immunotherapy

Chaisri

Chaicumpa

2018

BioMed Research International

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This narrative review article summarizes past and current technologies for generating antibodies for passive immunization/immunotherapy. Contemporary DNA and protein technologies have facilitated the development of engineered therapeutic monoclonal antibodies in a variety of formats according to the required effector functions. Chimeric, humanized, and human monoclonal antibodies to antigenic/epitopic myriads with less immunogenicity than animal-derived antibodies in human recipients can be produced in vitro. Immunotherapy with ready-to-use antibodies has gained wide acceptance as a powerful treatment against both infectious and noninfectious diseases. Influenza, a highly contagious disease, precipitates annual epidemics and occasional pandemics, resulting in high health and economic burden worldwide. Currently available drugs are becoming less and less effective against this rapidly mutating virus. Alternative treatment strategies are needed, particularly for individuals at high risk for severe morbidity. In a setting where vaccines are not yet protective or available, human antibodies that are broadly effective against various influenza subtypes could be highly efficacious in lowering morbidity and mortality and controlling unprecedented epidemic/pandemic. Prototypes of human single-chain antibodies to several conserved proteins of influenza virus with no Fc portion (hence, no ADE effect in recipients) are available. These antibodies have high potential as a novel, safe, and effective anti-influenza agent.

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“…EBOV patients receive only palliative therapy. 4 Currently, there are no licensed vaccines or treatments available to combat EBOV disease and as such, research aimed at identifying targets for therapeutic intervention is of high priority. However, the classification of EBOV as a biosafety level 4 (BSL-4) pathogen greatly limits studies using a live virus.…”

Section: Introductionmentioning

confidence: 99%

Ebola Virus L Polymerase RdRp Sequence and Phylogenetic Analysis

gohary

Eissa

Elfiky

et al. 2019

Egypt. J. Biophys. Biomed. Engng.

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E BOLA Virus (EBOV) infection affects humans beings in the last four decades with the deadliest outbreak at 2015 in West Africa, leaving more than 10,000 deaths. The virus harms the liver of the patient when direct contact with contaminated body fluids or blood is occurred. L polymerase is one of the viral proteins responsible for the viral RNA replication. Inhibition of the viral polymerase succeeded is in stopping the infection of other viruses such as Hepatitis C Virus (HCV). National Center for Biotechnology Information (NCBI) protein database has 2123 sequences for L polymerase. In present research, the sequence and phylogenetic analysis are utilized to understand the non-redundant sequence coming from different countries. Based on the sequence similarity, the solved structure of vesicular stomatitis virus (PDB ID: 5A22) is used in this work to suggest the active site of the EBOV RdRp domain. Two newly released sequences (APT69557.1 and ALX33626.1 for the Sudan and Zaire, respectively) are based on the phylogenetic analysis, show interestingly a divert, mutation and distance from its subgroups suggesting a new emerged isoform of EBOV RdRp. The active site motif, GDN, would be targeted by polymerase inhibitors succeeded in other viruses to get stop the infection.

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Human transbodies to VP40 inhibit cellular egress of Ebola virus-like particles

Cited by 19 publications

References 33 publications

Intracellular Crosslinking of Filoviral Nucleoproteins with Xintrabodies Restricts Viral Packaging

Intracellular Crosslinking of Filoviral Nucleoproteins with Xintrabodies Restricts Viral Packaging

Evolution of Therapeutic Antibodies, Influenza Virus Biology, Influenza, and Influenza Immunotherapy

Ebola Virus L Polymerase RdRp Sequence and Phylogenetic Analysis

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