Potent neutralization of vaccinia virus by divergent murine antibodies targeting a common site of vulnerability in L1 protein

Thomas Kaever, Xiangzhi Meng, Michael H. Matho, Andrew Schlossman, Sheng Li, Inbal Sela-Culang, Yanay Ofran, Mark Buller, Ryan W. Crump, Scott Parker, April Frazier, Shane Crotty, Dirk M. Zajonc, Bjoern Peters, Yan Xiang

Research output: Contribution to journalArticle

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Abstract

Vaccinia virus (VACV) L1 is an important target for viral neutralization and has been included in multicomponent DNA or protein vaccines against orthopoxviruses. To further understand the protective mechanism of the anti-L1 antibodies, we generated five murine anti-L1 monoclonal antibodies (MAbs), which clustered into 3 distinct epitope groups. While two groups of anti-L1 failed to neutralize, one group of 3 MAbs potently neutralized VACV in an isotype- and complement-independent manner. This is in contrast to neutralizing antibodies against major VACV envelope proteins, such as H3, D8, or A27, which failed to completely neutralize VACV unless the antibodies are of complement-fixing isotypes and complement is present. Compared to nonneutralizing anti-L1 MAbs, the neutralization antibodies bound to the recombinant L1 protein with a significantly higher affinity and also could bind to virions. By using a variety of techniques, including the isolation of neutralization escape mutants, hydrogen/deuterium exchange mass spectrometry, and X-ray crystallography, the epitope of the neutralizing antibodies was mapped to a conformational epitope with Asp35 as the key residue. This epitope is similar to the epitope of 7D11, a previously described potent VACV neutralizing antibody. The epitope was recognized mainly by CDR1 and CDR2 of the heavy chain, which are highly conserved among antibodies recognizing the epitope. These antibodies, however, had divergent light-chain and heavychain CDR3 sequences. Our study demonstrates that the conformational L1 epitope with Asp35 is a common site of vulnerability for potent neutralization by a divergent group of antibodies.

Original languageEnglish (US)
Pages (from-to)11339-11355
Number of pages17
JournalJournal of Virology
Volume88
Issue number19
DOIs
StatePublished - 2014

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Vaccinia virus
neutralization
epitopes
Epitopes
antibodies
Antibodies
mice
Proteins
proteins
Neutralizing Antibodies
neutralizing antibodies
monoclonal antibodies
complement
Monoclonal Antibodies
Orthopoxvirus
Viral Envelope Proteins
deuterium
Deuterium
X Ray Crystallography
isolation techniques

ASJC Scopus subject areas

  • Immunology
  • Virology

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Potent neutralization of vaccinia virus by divergent murine antibodies targeting a common site of vulnerability in L1 protein. / Kaever, Thomas; Meng, Xiangzhi; Matho, Michael H.; Schlossman, Andrew; Li, Sheng; Sela-Culang, Inbal; Ofran, Yanay; Buller, Mark; Crump, Ryan W.; Parker, Scott; Frazier, April; Crotty, Shane; Zajonc, Dirk M.; Peters, Bjoern; Xiang, Yan.

In: Journal of Virology, Vol. 88, No. 19, 2014, p. 11339-11355.

Research output: Contribution to journalArticle

Kaever, T, Meng, X, Matho, MH, Schlossman, A, Li, S, Sela-Culang, I, Ofran, Y, Buller, M, Crump, RW, Parker, S, Frazier, A, Crotty, S, Zajonc, DM, Peters, B & Xiang, Y 2014, 'Potent neutralization of vaccinia virus by divergent murine antibodies targeting a common site of vulnerability in L1 protein', Journal of Virology, vol. 88, no. 19, pp. 11339-11355. https://doi.org/10.1128/JVI.01491-14
Kaever, Thomas ; Meng, Xiangzhi ; Matho, Michael H. ; Schlossman, Andrew ; Li, Sheng ; Sela-Culang, Inbal ; Ofran, Yanay ; Buller, Mark ; Crump, Ryan W. ; Parker, Scott ; Frazier, April ; Crotty, Shane ; Zajonc, Dirk M. ; Peters, Bjoern ; Xiang, Yan. / Potent neutralization of vaccinia virus by divergent murine antibodies targeting a common site of vulnerability in L1 protein. In: Journal of Virology. 2014 ; Vol. 88, No. 19. pp. 11339-11355.
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AU - Parker, Scott

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