Immunosilencing a Highly Immunogenic Protein Trimerization Domain

Many therapeutic proteins and protein subunit vaccines contain heterologous trimerization domains, such as the widely used GCN4-based isoleucine zipper (IZ) and the T4 bacteriophage fibritin foldon (Fd) trimerization domains. We found that these domains induced potent anti-IZ or anti-Fd antibody res...

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Published inThe Journal of biological chemistry Vol. 290; no. 12; pp. 7436 - 7442
Main Authors Sliepen, Kwinten, van Montfort, Thijs, Melchers, Mark, Isik, Gözde, Sanders, Rogier W.
Format Journal Article
LanguageEnglish
Published United States Elsevier Inc 20.03.2015
American Society for Biochemistry and Molecular Biology
Subjects
Amino Acid Sequence
Animals
Enzyme-Linked Immunosorbent Assay
HEK293 Cells
human immunodeficiency virus (HIV)
Humans
humoral response
Immunology
influenza
Mice
Molecular Sequence Data
Mutagenesis
Polymerization
protein engineering
Proteins - chemistry
Proteins - genetics
Proteins - immunology
Rabbits
Rats
Rats, Wistar
vaccine
influenza
protein engineering
vaccine
human immunodeficiency virus (HIV)
humoral response
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Summary:Many therapeutic proteins and protein subunit vaccines contain heterologous trimerization domains, such as the widely used GCN4-based isoleucine zipper (IZ) and the T4 bacteriophage fibritin foldon (Fd) trimerization domains. We found that these domains induced potent anti-IZ or anti-Fd antibody responses in animals when fused to an HIV-1 envelope glycoprotein (Env) immunogen. To dampen IZ-induced responses, we constructed an IZ domain containing four N-linked glycans (IZN4) to shield the underlying protein surface. When fused to two different vaccine antigens, HIV-1 Env and influenza hemagglutinin (HA), IZN4 strongly reduced the antibody responses against the IZ, but did not affect the antibody titers against Env or HA. Silencing of immunogenic multimerization domains with glycans might be relevant for therapeutic proteins and protein vaccines. Background: Trimerization domains are commonly used to stabilize trimeric protein vaccines and therapeutics. Results: The GCN4-based isoleucine zipper domain induces strong antibody responses in vivo but this can be overcome by introducing glycans. Conclusion: Appropriately positioned glycans can effectively immunosilence the GCN4-based trimerization domain. Significance: Immunosilencing trimerization domains could be important for the exploitation of trimerization domains in protein vaccines and therapeutics.
Bibliography:These authors contributed equally to this work.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.M114.620534