Quantification of Lyssavirus-Neutralizing Antibodies Using Vesicular Stomatitis Virus Pseudotype Particles.

Sarah Moeschler,Karl-Klaus Conzelmann,Beate Krämer,Gert Zimmer,Samira Locher

doi:10.3390/v8090254

Sarah Moeschler, Karl-Klaus Conzelmann + Show 3 more

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https://doi.org/10.3390/v8090254

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Abstract

Rabies is a highly fatal zoonotic disease which is primarily caused by rabies virus (RABV) although other members of the genus Lyssavirus can cause rabies as well. As yet, 14 serologically and genetically diverse lyssaviruses have been identified, mostly in bats. To assess the quality of rabies vaccines and immunoglobulin preparations, virus neutralization tests with live RABV are performed in accordance with enhanced biosafety standards. In the present work, a novel neutralization test is presented which takes advantage of a modified vesicular stomatitis virus (VSV) from which the glycoprotein G gene has been deleted and replaced by reporter genes. This single-cycle virus was trans-complemented with RABV envelope glycoprotein. Neutralization of this pseudotype virus with RABV reference serum or immune sera from vaccinated mice showed a strong correlation with the rapid fluorescent focus inhibition test (RFFIT). Importantly, pseudotype viruses containing the envelope glycoproteins of other lyssaviruses were neutralized by reference serum to a significantly lesser extent or were not neutralized at all. Taken together, a pseudotype virus system has been successfully developed which allows the safe, fast, and sensitive detection of neutralizing antibodies directed against different lyssaviruses.

Highlights

Rabies is a fatal neurological disorder in humans and other mammalian species which is caused by rabies virus (RABV), the prototype member of the genus Lyssavirus within the family Rhabdoviridae [1].The Lyssavirus genus currently contains 14 classified and one proposed species which are classified into distinct phylogenetic groups [2,3,4,5,6,7,8]
Group 1 consists of RABV, Khujand virus (KHUV), Australian bat lyssavirus (ABLV), European bat lyssavirus type 1 (EBLV-1), European bat lyssavirus type 2 (EBLV-2), Bokeloh bat lyssavirus (BBLV), Aravan virus (ARAV), Duvenhage virus (DUVV), and Irkut virus (IRKV)
It has been previously shown that vesicular stomatitis virus (VSV) lacking the glycoprotein G (VSV∆G) can be efficiently trans-complemented with the VSV G protein using a transgenic cell line [22]

Summary

Introduction

Rabies is a fatal neurological disorder in humans and other mammalian species which is caused by rabies virus (RABV), the prototype member of the genus Lyssavirus within the family Rhabdoviridae [1]. The Lyssavirus genus currently contains 14 classified and one proposed species which are classified into distinct phylogenetic groups [2,3,4,5,6,7,8]. Group 1 consists of RABV, Khujand virus (KHUV), Australian bat lyssavirus (ABLV), European bat lyssavirus type 1 (EBLV-1), European bat lyssavirus type 2. (EBLV-2), Bokeloh bat lyssavirus (BBLV), Aravan virus (ARAV), Duvenhage virus (DUVV), and Irkut virus (IRKV). For Ikoma and Lleida bat lyssaviruses (IKOV and LLEBV, respectively) the establishment of a novel group 4 has been proposed [7,8]. Except MOKV, which has been found in shrews and cats, all other lyssaviruses have their natural reservoir in bats

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