Cleavage-independent HIV-1 trimers from CHO cell lines elicit robust autologous tier 2 neutralizing antibodies
Cleavage-independent HIV-1 trimers from CHO cell lines elicit robust autologous tier 2 neutralizing antibodies
Native flexibly linked (NFL) HIV-1 envelope glycoprotein (Env) trimers are cleavage-independent and display a native-like, well-folded conformation that preferentially displays broadly neutralizing determinants. The NFL platform simplifies large-scale production of Env by eliminating the need to co-transfect the precursor-cleaving protease, furin that is required by the cleavage-dependent SOSIP trimers. Here, we report the development of a CHO-M cell line that expressed BG505 NFL trimers at a high level of homogeneity and yields of ~1.8 g/l. BG505 NFL trimers purified by single-step lectin-affinity chromatography displayed a native-like closed structure, efficient recognition by trimer-preferring bNAbs, no recognition by non-neutralizing CD4 binding site-directed and V3-directed antibodies, long-term stability, and proper N-glycan processing. Following negative-selection, formulation in ISCOMATRIX adjuvant and inoculation into rabbits, the trimers rapidly elicited potent autologous tier 2 neutralizing antibodies. These antibodies targeted the N-glycan "hole" naturally present on the BG505 Env proximal to residues at positions 230, 241, and 289. The BG505 NFL trimers that did not expose V3 in vitro, elicited low-to-no tier 1 virus neutralization in vivo, indicating that they remained intact during the immunization process, not exposing V3. In addition, BG505 NFL and BG505 SOSIP trimers expressed from 293F cells, when formulated in Adjuplex adjuvant, elicited equivalent BG505 tier 2 autologous neutralizing titers. These titers were lower in potency when compared to the titers elicited by CHO-M cell derived trimers. In addition, increased neutralization of tier 1 viruses was detected. Taken together, these data indicate that both adjuvant and cell-type expression can affect the elicitation of tier 2 and tier 1 neutralizing responses in vivo.
Adjuvants, HIV-1, Immunogenicity, Immunologic, Recombinant protein expression, Vaccines, Viral fusion proteins
Bale, Shridhar
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Martiné, Alexandra
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Wilson, Richard
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Behrens, Anna Janina
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Fourn, Valérie Le
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Val, Natalia de
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Sharma, Shailendra K.
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Tran, Karen
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Torres, Jonathan L.
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Girod, Pierre Alain
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Ward, Andrew B.
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Crispin, Max
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Wyatt, Richard T.
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Bale, Shridhar
2b36dee6-de09-4f14-93ee-36675753434a
Martiné, Alexandra
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Wilson, Richard
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Behrens, Anna Janina
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Fourn, Valérie Le
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Val, Natalia de
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Sharma, Shailendra K.
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Tran, Karen
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Torres, Jonathan L.
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Girod, Pierre Alain
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Ward, Andrew B.
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Crispin, Max
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Wyatt, Richard T.
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Bale, Shridhar, Martiné, Alexandra, Wilson, Richard, Behrens, Anna Janina, Fourn, Valérie Le, Val, Natalia de, Sharma, Shailendra K., Tran, Karen, Torres, Jonathan L., Girod, Pierre Alain, Ward, Andrew B., Crispin, Max and Wyatt, Richard T.
(2018)
Cleavage-independent HIV-1 trimers from CHO cell lines elicit robust autologous tier 2 neutralizing antibodies.
Frontiers in Immunology, 9 (MAY), [1116].
(doi:10.3389/fimmu.2018.01116).
Abstract
Native flexibly linked (NFL) HIV-1 envelope glycoprotein (Env) trimers are cleavage-independent and display a native-like, well-folded conformation that preferentially displays broadly neutralizing determinants. The NFL platform simplifies large-scale production of Env by eliminating the need to co-transfect the precursor-cleaving protease, furin that is required by the cleavage-dependent SOSIP trimers. Here, we report the development of a CHO-M cell line that expressed BG505 NFL trimers at a high level of homogeneity and yields of ~1.8 g/l. BG505 NFL trimers purified by single-step lectin-affinity chromatography displayed a native-like closed structure, efficient recognition by trimer-preferring bNAbs, no recognition by non-neutralizing CD4 binding site-directed and V3-directed antibodies, long-term stability, and proper N-glycan processing. Following negative-selection, formulation in ISCOMATRIX adjuvant and inoculation into rabbits, the trimers rapidly elicited potent autologous tier 2 neutralizing antibodies. These antibodies targeted the N-glycan "hole" naturally present on the BG505 Env proximal to residues at positions 230, 241, and 289. The BG505 NFL trimers that did not expose V3 in vitro, elicited low-to-no tier 1 virus neutralization in vivo, indicating that they remained intact during the immunization process, not exposing V3. In addition, BG505 NFL and BG505 SOSIP trimers expressed from 293F cells, when formulated in Adjuplex adjuvant, elicited equivalent BG505 tier 2 autologous neutralizing titers. These titers were lower in potency when compared to the titers elicited by CHO-M cell derived trimers. In addition, increased neutralization of tier 1 viruses was detected. Taken together, these data indicate that both adjuvant and cell-type expression can affect the elicitation of tier 2 and tier 1 neutralizing responses in vivo.
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fimmu-09-01116
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Accepted/In Press date: 3 May 2018
e-pub ahead of print date: 24 May 2018
Keywords:
Adjuvants, HIV-1, Immunogenicity, Immunologic, Recombinant protein expression, Vaccines, Viral fusion proteins
Identifiers
Local EPrints ID: 421520
URI: http://eprints.soton.ac.uk/id/eprint/421520
ISSN: 1664-3224
PURE UUID: 49365cbc-408d-4713-a1f7-83c921885839
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Date deposited: 14 Jun 2018 16:30
Last modified: 16 Mar 2024 04:30
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Contributors
Author:
Shridhar Bale
Author:
Alexandra Martiné
Author:
Richard Wilson
Author:
Anna Janina Behrens
Author:
Valérie Le Fourn
Author:
Natalia de Val
Author:
Shailendra K. Sharma
Author:
Karen Tran
Author:
Jonathan L. Torres
Author:
Pierre Alain Girod
Author:
Andrew B. Ward
Author:
Richard T. Wyatt
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