generation-recombinant-arenavirus-for-vaccine-development-fda

Generation of Recombinant Arenavirus for Vaccine Development in FDA-Approved Vero Cells

The development and implementation of arenavirus reverse genetics represents a significant breakthrough in the arenavirus field  4. The use of cell-based ...

University of Rochester School of Medicine and Dentistry

Arenaviruses merit significant interest because several family members are etiological agents of severe hemorrhagic fevers, representing a major burden to public health. Currently, there are no FDA-licensed vaccines against arenaviruses and the only available antiviral therapy is limited to the use of ribavirin that is partially effective. Arenavirus nucleoprotein (NP) is found associated with the genomic RNA forming the viral ribonucleoproteins (vRNPs) that together with the polymerase (L) direct viral replication and transcription. Virion formation requires the recruitment of vRNPs into budding sites, a process in which the arenavirus matrix-like protein (Z) plays a major role. Therefore, proper NP-NP and NP-Z interactions are required for the generation of infectious progeny. In this work we demonstrate the role of the amino acid residue D471 in the self-association of lymphocytic choriomeningitis virus nucleoprotein (LCMV-NP). Amino acid substitutions at this position abrogate NP oligomerization, affecting its ability to mediate replication and transcription of a minigenome reporter plasmid. However, its ability to interact with the Z protein, counteract the cellular interferon response and bind to dsRNA analogs was retained. Additionally, we also document the dominant negative effect of D471G mutation on viral infection, suggesting that NP self-association is an excellent target for the development of new antivirals against arenaviruses.

D471G mutation in LCMV-NP affects its ability to self-associate and results in a dominant negative effect in viral RNA synthesis.

Arenavirus m&eacute;ritent beaucoup d&#39;int&eacute;r&ecirc;t parce que plusieurs membres de la famille sont des agents &eacute;tiologiques des fi&egrave;vres h&eacute;morragiques s&eacute;v&egrave;res, ce qui repr&eacute;sente un lourd fardeau pour la sant&eacute; publique. Actuellement, il n&#39;y a aucun vaccins homologu&eacute;s FDA contre arenavirus et la th&eacute;rapie antivirale uniquement disponible est limit&eacute;e &agrave; l&#39;utilisation de la ribavirine qui est partiellement efficace. Ar&eacute;navirus nucl&eacute;oprot&eacute;ine (NP) se trouve associ&eacute;e &agrave; l&#39;ARN g&eacute;nomique formant les ribonucl&eacute;oprot&eacute;ines virales (vRNPs) qui, avec la polym&eacute;rase (L), directement la r&eacute;plication virale et la transcription. Virion formation n&eacute;cessite le recrutement de vRNPs dans les sites en herbe, un processus dans lequel la prot&eacute;ine de matrice arenavirus (Z) joue un r&ocirc;le majeur. Donc, bonne interactions NP-NP NP-Z sont exig&eacute;es pour la g&eacute;n&eacute;ration de descendants infectieuses. Dans ce travail, nous d&eacute;montrons le r&ocirc;le du r&eacute;sidu d&#39;acide amin&eacute; D471 dans l&#39;auto-association de nucl&eacute;oprot&eacute;ine de virus de la choriom&eacute;ningite lymphocytaire (CML-NP). Des substitutions d&#39;acides amin&eacute;s &agrave; cette position abroger l&#39;oligom&eacute;risation NP, affectant sa capacit&eacute; de provoquer la r&eacute;plication et la transcription d&#39;un plasmide de journaliste de minigenome. Cependant, sa capacit&eacute; &agrave; interagir avec la prot&eacute;ine Z, contrecarrer la r&eacute;ponse interf&eacute;ron cellulaire et se lient aux analogues de l&#39;ARNdb est maintenue. En outre, nous documentons &eacute;galement l&#39;effet dominant n&eacute;gatif de D471G mutation sur infection virale, sugg&eacute;rant que l&#39;auto-association NP est une excellente cible pour le d&eacute;veloppement de nouveaux agents antiviraux contre ar&eacute;navirus.

CML-NP D471G mutation affecte sa capacit&eacute; d&#39;auto-association et se traduit par un effet dominant n&eacute;gatif dans la synth&egrave;se de l&#39;ARN viral.

Arenaviruses meritano un interesse significativo perch&eacute; diversi membri della famiglia sono agenti eziologici di febbri emorragiche gravi, che rappresentano un onere maggiore per la salute pubblica. Attualmente, non ci sono nessuna licenza FDA-vaccini contro la arenaviruses e la terapia antivirale solo disponibile &egrave; limitata all&#39;uso di ribavirina che sono parzialmente efficace. Arenavirus nucleoproteina (NP) si trova associata con il RNA genomico formando il ribonucleoproteins virale (vRNPs) che, insieme con la polimerasi (L) diretta trascrizione e replicazione virale. Formazione del virione richiede l&#39;assunzione di vRNPs in siti in erba, un processo in cui la proteina di matrice come arenavirus (Z) svolge un ruolo importante. Pertanto, corrette interazioni NP NP e NP-Z sono necessarie per la generazione della progenie infettiva. In questo lavoro ci dimostrano il ruolo del residuo dell&#39;amminoacido D471 in moltiplicava della nucleoproteina di virus della Coriomeningite linfocitaria (LCMV-NP). Aminoacidiche in questa posizione abrogare oligomerizzazione NP, che colpisce la sua capacit&agrave; di mediare la replica e la trascrizione di un plasmide del reporter minigenome. Tuttavia, la capacit&agrave; di interagire con la proteina Z, contrastare la risposta cellulare interferone e associare a dsRNA analoghi &egrave; stata mantenuta. Inoltre, abbiamo anche documentare l&#39;effetto dominante negativo della mutazione D471G su infezione virale, suggerendo che moltiplicava NP &egrave; un&#39;eccellente destinazione per lo sviluppo di nuovi farmaci antivirali contro arenaviruses.

Mutazione D471G in LCMV-NP influisce sulla sua capacit&agrave; di self-associate e si traduce in un effetto negativo dominante nella sintesi di RNA virale.

Benson Y.H. Cheng

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