Chimeric filoviruses for identification and characterization of monoclonal antibodies

Philipp A. Ilinykh, Xiaoli Shen, Andrew I. Flyak, Natalia Kuzmina, Thomas Ksiazek, James E. Crowe, Alexander Bukreyev

Research output: Contribution to journalArticle

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Abstract

Recent experiments suggest that some glycoprotein (GP)-specific monoclonal antibodies (MAbs) can protect experimental animals against the filovirus Ebola virus (EBOV). There is a need for isolation of MAbs capable of neutralizing multiple filoviruses. Antibody neutralization assays for filoviruses frequently use surrogate systems such as the rhabdovirus vesicular stomatitis Indiana virus (VSV), lentiviruses or gammaretroviruses with their envelope proteins replaced with EBOV GP or pseudotyped with EBOV GP. It is optimal for both screening and in-depth characterization of newly identified neutralizing MAbs to generate recombinant filoviruses that express a reporter fluorescent protein in order to more easily monitor and quantify the infection. Our study showed that unlike neutralization-sensitive chimeric VSV, authentic filoviruses are highly resistant to neutralization by MAbs. We used reverse genetics techniques to replace EBOV GP with its counterpart from the heterologous filoviruses Bundibugyo virus (BDBV), Sudan virus, and even Marburg virus and Lloviu virus, which belong to the heterologous genera in the filovirus family. This work resulted in generation of multiple chimeric filoviruses, demonstrating the ability of filoviruses to tolerate swapping of the envelope protein. The sensitivity of chimeric filoviruses to neutralizing MAbs was similar to that of authentic biologically derived filoviruses with the same GP. Moreover, disabling the expression of the secreted GP (sGP) resulted in an increased susceptibility of an engineered virus to the BDBV52 MAb isolated from a BDBV survivor, suggesting a role for sGP in evasion of antibody neutralization in the context of a human filovirus infection.

Original languageEnglish (US)
Pages (from-to)3890-3901
Number of pages12
JournalJournal of Virology
Volume90
Issue number8
DOIs
StatePublished - Apr 1 2016

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Filoviridae
Ebolavirus
monoclonal antibodies
Glycoproteins
Monoclonal Antibodies
Viruses
Vesicular stomatitis Indiana virus
glycoproteins
Neutralizing Antibodies
neutralization
Marburgvirus
viruses
Gammaretrovirus
Rhabdoviridae
Genetic Techniques
Reverse Genetics
Lentivirus
Proteins
Sudan
Antibodies

ASJC Scopus subject areas

  • Immunology
  • Virology

Cite this

Chimeric filoviruses for identification and characterization of monoclonal antibodies. / Ilinykh, Philipp A.; Shen, Xiaoli; Flyak, Andrew I.; Kuzmina, Natalia; Ksiazek, Thomas; Crowe, James E.; Bukreyev, Alexander.

In: Journal of Virology, Vol. 90, No. 8, 01.04.2016, p. 3890-3901.

Research output: Contribution to journalArticle

Ilinykh, Philipp A. ; Shen, Xiaoli ; Flyak, Andrew I. ; Kuzmina, Natalia ; Ksiazek, Thomas ; Crowe, James E. ; Bukreyev, Alexander. / Chimeric filoviruses for identification and characterization of monoclonal antibodies. In: Journal of Virology. 2016 ; Vol. 90, No. 8. pp. 3890-3901.
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