title: Chimeric HIV-1 envelope glycoproteins with potent intrinsic granulocyte-macrophage colony-stimulating factor (GM-CSF) activity.
abstract: HIV-1 acquisition can be prevented by broadly neutralizing antibodies (BrNAbs) that target the envelope glycoprotein complex (Env). An ideal vaccine should therefore be able to induce BrNAbs that can provide immunity over a prolonged period of time, but the low intrinsic immunogenicity of HIV-1 Env makes the elicitation of such BrNAbs challenging. Co-stimulatory molecules can increase the immunogenicity of Env and we have engineered a soluble chimeric Env trimer with an embedded granulocyte-macrophage colony-stimulating factor (GM-CSF) domain. This chimeric molecule induced enhanced B and helper T cell responses in mice compared to Env without GM-CSF. We studied whether we could optimize the activity of the embedded GM-CSF as well as the antigenic structure of the Env component of the chimeric molecule. We assessed the effect of truncating GM-CSF, removing glycosylation-sites in GM-CSF, and adjusting the linker length between GM-CSF and Env. One of our designed Env(GM-CSF) chimeras improved GM-CSF-dependent cell proliferation by 6-fold, reaching the same activity as soluble recombinant GM-CSF. In addition, we incorporated GM-CSF into a cleavable Env trimer and found that insertion of GM-CSF did not compromise Env cleavage, while Env cleavage did not compromise GM-CSF activity. Importantly, these optimized Env(GM-CSF) proteins were able to differentiate human monocytes into cells with a macrophage-like phenotype. Chimeric Env(GM-CSF) should be useful for improving humoral immunity against HIV-1 and these studies should inform the design of other chimeric proteins.
authors.affiliation: Department of Medical Microbiology, Laboratory of Experimental Virology, Center for Infection and Immunity Amsterdam, University of Amsterdam, Amsterdam, The Netherlands.
authors.forename: Gözde
authors.initials: G
authors.lastname: Isik
authors.forename: Thijs
authors.initials: T
authors.lastname: van Montfort
authors.forename: Maikel
authors.initials: M
authors.lastname: Boot
authors.forename: Viviana
authors.initials: V
authors.lastname: Cobos Jiménez
authors.forename: Neeltje A
authors.initials: NA
authors.lastname: Kootstra
authors.forename: Rogier W
authors.initials: RW
authors.lastname: Sanders
mesh_terms: D057134:Antibodies, Neutralizing; D015704:CD4 Antigens / Q000378:metabolism; D002460:Cell Line; D000939:Epitopes / Q000276:immunology; D023061:Gene Order; D006031:Glycosylation; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D015483:HIV Antibodies / Q000276:immunology / Q000378:metabolism; D015497:HIV-1 / Q000235:genetics* / Q000276:immunology; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology / Q000378:metabolism; D009000:Monocytes / Q000276:immunology / Q000378:metabolism; D009154:Mutation; D011485:Protein Binding / Q000276:immunology; D054730:Protein Interaction Domains and Motifs; D059748:Proteolysis; D011993:Recombinant Fusion Proteins; D054299:env Gene Products, Human Immunodeficiency Virus / Q000235:genetics* / Q000276:immunology / Q000378:metabolism
chemical_list: D057134:Antibodies, Neutralizing; D015704:CD4 Antigens; D000939:Epitopes; D015483:HIV Antibodies; D007074:Immunoglobulin G; D011993:Recombinant Fusion Proteins; D054299:env Gene Products, Human Immunodeficiency Virus; D016178:Granulocyte-Macrophage Colony-Stimulating Factor
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