Idioma:

A native-like SOSIP.664 trimer based on an HIV-1 subtype B env gene

Pugach, Pavel ; Ozorowski, Gabriel ; Cupo, Albert ; Ringe, Rajesh ; Yasmeen, Anila ; de Val, Natalia ; Derking, Ronald ; Kim, Helen J ; Korzun, Jacob ; Golabek, Michael ; de Los Reyes, Kevin ; Ketas, Thomas J ; Julien, Jean-Philippe ; Burton, Dennis R ; Wilson, Ian A ; Sanders, Rogier W ; Klasse, P J ; Ward, Andrew B ; Moore, John P Doms, R. W.

Journal of virology, 2015-03, Vol.89 (6), p.3380-3395 [Periódico revisado por pares]

United States: American Society for Microbiology

Texto completo disponível

Citações Citado por

Enviar para

Título:
A native-like SOSIP.664 trimer based on an HIV-1 subtype B env gene
Autor: Pugach, Pavel ; Ozorowski, Gabriel ; Cupo, Albert ; Ringe, Rajesh ; Yasmeen, Anila ; de Val, Natalia ; Derking, Ronald ; Kim, Helen J ; Korzun, Jacob ; Golabek, Michael ; de Los Reyes, Kevin ; Ketas, Thomas J ; Julien, Jean-Philippe ; Burton, Dennis R ; Wilson, Ian A ; Sanders, Rogier W ; Klasse, P J ; Ward, Andrew B ; Moore, John P
Doms, R. W.
Assuntos: AIDS Vaccines - chemistry ; AIDS Vaccines - genetics ; AIDS Vaccines - immunology ; AIDS Vaccines - isolation & purification ; env Gene Products, Human Immunodeficiency Virus - chemistry ; env Gene Products, Human Immunodeficiency Virus - genetics ; env Gene Products, Human Immunodeficiency Virus - immunology ; env Gene Products, Human Immunodeficiency Virus - isolation & purification ; Epitopes - chemistry ; Epitopes - genetics ; Epitopes - immunology ; HIV Infections - virology ; HIV-1 - chemistry ; HIV-1 - classification ; HIV-1 - genetics ; HIV-1 - immunology ; Human immunodeficiency virus 1 ; Humans ; Protein Multimerization ; Vaccines and Antiviral Agents
É parte de: Journal of virology, 2015-03, Vol.89 (6), p.3380-3395
Notas: ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
Present address: Jean-Philippe Julien, Program in Molecular Structure and Function, The Hospital for Sick Children Research Institute and Departments of Biochemistry and Immunology, University of Toronto, Toronto, Ontario, Canada.
Citation Pugach P, Ozorowski G, Cupo A, Ringe R, Yasmeen A, de Val N, Derking R, Kim HJ, Korzun J, Golabek M, de los Reyes K, Ketas TJ, Julien J-P, Burton DR, Wilson IA, Sanders RW, Klasse PJ, Ward AB, Moore JP. 2015. A native-like SOSIP.664 trimer based on an HIV-1 subtype B env gene. J Virol 89:3380–3395. doi:10.1128/JVI.03473-14.
P.P. and G.O. contributed equally to this article.
Descrição: Recombinant trimeric mimics of the human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein (Env) spike should expose as many epitopes as possible for broadly neutralizing antibodies (bNAbs) but few, if any, for nonneutralizing antibodies (non-NAbs). Soluble, cleaved SOSIP.664 gp140 trimers based on the subtype A strain BG505 approach this ideal and are therefore plausible vaccine candidates. Here, we report on the production and in vitro properties of a new SOSIP.664 trimer derived from a subtype B env gene, B41, including how to make this protein in low-serum media without proteolytic damage (clipping) to the V3 region. We also show that nonclipped trimers can be purified successfully via a positive-selection affinity column using the bNAb PGT145, which recognizes a quaternary structure-dependent epitope at the trimer apex. Negative-stain electron microscopy imaging shows that the purified, nonclipped, native-like B41 SOSIP.664 trimers contain two subpopulations, which we propose represent an equilibrium between the fully closed and a more open conformation. The latter is different from the fully open, CD4 receptor-bound conformation and may represent an intermediate state of the trimer. This new subtype B trimer adds to the repertoire of native-like Env proteins that are suitable for immunogenicity and structural studies. The cleaved, trimeric envelope protein complex is the only neutralizing antibody target on the HIV-1 surface. Many vaccine strategies are based on inducing neutralizing antibodies. For HIV-1, one approach involves using recombinant, soluble protein mimics of the native trimer. At present, the only reliable way to make native-like, soluble trimers in practical amounts is via the introduction of specific sequence changes that confer stability on the cleaved form of Env. The resulting proteins are known as SOSIP.664 gp140 trimers, and the current paradigm is based on the BG505 subtype A env gene. Here, we describe the production and characterization of a SOSIP.664 protein derived from a subtype B gene (B41), together with a simple, one-step method to purify native-like trimers by affinity chromatography with a trimer-specific bNAb, PGT145. The resulting trimers will be useful for structural and immunogenicity experiments aimed at devising ways to make an effective HIV-1 vaccine.
Editor: United States: American Society for Microbiology
Idioma: Inglês

Links

View this record in MEDLINE/PubMed

Voltar para lista de resultados

Realização: Logos de Redes Sociais:

A native-like SOSIP.664 trimer based on an HIV-1 subtype B env gene

United States: American Society for Microbiology

Buscando em bases de dados remotas. Favor aguardar.