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-...
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Format: | Article |
Language: | English |
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Frontiers Media S.A.
2018-05-01
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Series: | Frontiers in Immunology |
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Online Access: | https://www.frontiersin.org/article/10.3389/fimmu.2018.01116/full |
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doaj-08399d971e284ca3bf665ab93adae2a1 |
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record_format |
Article |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Shridhar Bale Alexandra Martiné Richard Wilson Anna-Janina Behrens Valérie Le Fourn Natalia de Val Natalia de Val Shailendra K. Sharma Karen Tran Jonathan L. Torres Jonathan L. Torres Jonathan L. Torres Pierre-Alain Girod Andrew B. Ward Andrew B. Ward Andrew B. Ward Max Crispin Max Crispin Max Crispin Richard T. Wyatt Richard T. Wyatt Richard T. Wyatt |
spellingShingle |
Shridhar Bale Alexandra Martiné Richard Wilson Anna-Janina Behrens Valérie Le Fourn Natalia de Val Natalia de Val Shailendra K. Sharma Karen Tran Jonathan L. Torres Jonathan L. Torres Jonathan L. Torres Pierre-Alain Girod Andrew B. Ward Andrew B. Ward Andrew B. Ward Max Crispin Max Crispin Max Crispin Richard T. Wyatt Richard T. Wyatt Richard T. Wyatt Cleavage-Independent HIV-1 Trimers From CHO Cell Lines Elicit Robust Autologous Tier 2 Neutralizing Antibodies Frontiers in Immunology HIV-1 immunogenicity viral fusion proteins vaccines recombinant protein expression adjuvants |
author_facet |
Shridhar Bale Alexandra Martiné Richard Wilson Anna-Janina Behrens Valérie Le Fourn Natalia de Val Natalia de Val Shailendra K. Sharma Karen Tran Jonathan L. Torres Jonathan L. Torres Jonathan L. Torres Pierre-Alain Girod Andrew B. Ward Andrew B. Ward Andrew B. Ward Max Crispin Max Crispin Max Crispin Richard T. Wyatt Richard T. Wyatt Richard T. Wyatt |
author_sort |
Shridhar Bale |
title |
Cleavage-Independent HIV-1 Trimers From CHO Cell Lines Elicit Robust Autologous Tier 2 Neutralizing Antibodies |
title_short |
Cleavage-Independent HIV-1 Trimers From CHO Cell Lines Elicit Robust Autologous Tier 2 Neutralizing Antibodies |
title_full |
Cleavage-Independent HIV-1 Trimers From CHO Cell Lines Elicit Robust Autologous Tier 2 Neutralizing Antibodies |
title_fullStr |
Cleavage-Independent HIV-1 Trimers From CHO Cell Lines Elicit Robust Autologous Tier 2 Neutralizing Antibodies |
title_full_unstemmed |
Cleavage-Independent HIV-1 Trimers From CHO Cell Lines Elicit Robust Autologous Tier 2 Neutralizing Antibodies |
title_sort |
cleavage-independent hiv-1 trimers from cho cell lines elicit robust autologous tier 2 neutralizing antibodies |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Immunology |
issn |
1664-3224 |
publishDate |
2018-05-01 |
description |
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. |
topic |
HIV-1 immunogenicity viral fusion proteins vaccines recombinant protein expression adjuvants |
url |
https://www.frontiersin.org/article/10.3389/fimmu.2018.01116/full |
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doaj-08399d971e284ca3bf665ab93adae2a12020-11-24T22:25:12ZengFrontiers Media S.A.Frontiers in Immunology1664-32242018-05-01910.3389/fimmu.2018.01116372964Cleavage-Independent HIV-1 Trimers From CHO Cell Lines Elicit Robust Autologous Tier 2 Neutralizing AntibodiesShridhar Bale0Alexandra Martiné1Richard Wilson2Anna-Janina Behrens3Valérie Le Fourn4Natalia de Val5Natalia de Val6Shailendra K. Sharma7Karen Tran8Jonathan L. Torres9Jonathan L. Torres10Jonathan L. Torres11Pierre-Alain Girod12Andrew B. Ward13Andrew B. Ward14Andrew B. Ward15Max Crispin16Max Crispin17Max Crispin18Richard T. Wyatt19Richard T. Wyatt20Richard T. Wyatt21Department of Immunology and Microbiology, The Scripps Research Institute, La Jolla, CA, United StatesSelexis SA, Geneva, SwitzerlandIAVI Neutralizing Antibody Center, The Scripps Research Institute, La Jolla, CA, United StatesDepartment of Biochemistry, Oxford Glycobiology Institute, University of Oxford, Oxford, United KingdomSelexis SA, Geneva, SwitzerlandIAVI Neutralizing Antibody Center, The Scripps Research Institute, La Jolla, CA, United StatesDepartment of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, CA, United StatesIAVI Neutralizing Antibody Center, The Scripps Research Institute, La Jolla, CA, United StatesIAVI Neutralizing Antibody Center, The Scripps Research Institute, La Jolla, CA, United StatesIAVI Neutralizing Antibody Center, The Scripps Research Institute, La Jolla, CA, United StatesDepartment of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, CA, United StatesCenter for HIV/AIDS Vaccine Immunology and Immunogen Discovery, The Scripps Research Institute, La Jolla, CA, United StatesSelexis SA, Geneva, SwitzerlandIAVI Neutralizing Antibody Center, The Scripps Research Institute, La Jolla, CA, United StatesDepartment of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, CA, United StatesCenter for HIV/AIDS Vaccine Immunology and Immunogen Discovery, The Scripps Research Institute, La Jolla, CA, United StatesDepartment of Immunology and Microbiology, The Scripps Research Institute, La Jolla, CA, United StatesDepartment of Biochemistry, Oxford Glycobiology Institute, University of Oxford, Oxford, United KingdomCentre for Biological Sciences, Institute of Life Sciences, University of Southampton, Southampton, United KingdomDepartment of Immunology and Microbiology, The Scripps Research Institute, La Jolla, CA, United StatesIAVI Neutralizing Antibody Center, The Scripps Research Institute, La Jolla, CA, United StatesCenter for HIV/AIDS Vaccine Immunology and Immunogen Discovery, The Scripps Research Institute, La Jolla, CA, United StatesNative 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.https://www.frontiersin.org/article/10.3389/fimmu.2018.01116/fullHIV-1immunogenicityviral fusion proteinsvaccinesrecombinant protein expressionadjuvants |