Dramatic potentiation of the antiviral activity of HIV antibodies by cholesterol conjugation
| Autor: | Jonathan K. Ball, Antimo Di Maro, Claudia De Lorenzo, Nigel J. Temperton, Alexander Ploss, Valeria Severino, Antonello Pessi, Richard A. Urbanowicz, Krzysztof Lacek, Alfredo Nicosia, Francesca Ferrara, Fulvia Troise, Alexander W. Tarr, Riccardo Cortese |
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| Přispěvatelé: | Lacek, K, Urbanowicz, Ra, Troise, F, DE LORENZO, Claudia, Severino, V, DI MARO, Antimo, Tarr, Aw, Ferrara, F, Ploss, A, Temperton, N, Ball, Jk, Nicosia, Alfredo, Cortese, Riccardo, Pessi, A., Lacek, K., Urbanowicz, R. A., Troise, F., De Lorenzo, C., Severino, V., Tarr, A. W., Ferrara, F., Ploss, A., Temperton, N., Ball, J. K., Nicosia, A., Cortese, R. |
| Rok vydání: | 2014 |
| Předmět: |
Immunology
Biology HIV Antibodies Gp41 Biochemistry Membrane Fusion Epitope Molecular Evolution Affinity maturation Antibody Engineering Neutralization Tests Humans Viral Immunology Molecular Biology QR355 Human Immunodeficiency Virus (HIV) Lipid Raft Cell Membrane Lipid bilayer fusion Antibodies Monoclonal Cell Biology Viral membrane Fusion protein Antibodies Neutralizing 3. Good health Cholesterol HEK293 Cells Virus Entry biology.protein HIV-1 Paratope Antibody |
| Zdroj: | The Journal of Biological Chemistry |
| ISSN: | 1083-351X 0021-9258 |
| Popis: | Background: Some HIV-neutralizing antibodies have an antigen-binding site with dual specificity, for the plasma membrane and a viral epitope. Results: We engineered membrane affinity outside the antigen-binding site by conjugating cholesterol, with concomitantly increased antiviral potency. Conclusion: A natural mechanism dependent on affinity maturation is mimicked by conjugation of a lipid. Significance: This is a general strategy to boost the potency of antiviral antibodies. The broadly neutralizing antibodies HIV 2F5 and 4E10, which bind to overlapping epitopes in the membrane-proximal external region of the fusion protein gp41, have been proposed to use a two-step mechanism for neutralization; first, they bind and preconcentrate at the viral membrane through their long, hydrophobic CDRH3 loops, and second, they form a high affinity complex with the protein epitope. Accordingly, mutagenesis of the CDRH3 can abolish their neutralizing activity, with no change in the affinity for the peptide epitope. We show here that we can mimic this mechanism by conjugating a cholesterol group outside of the paratope of an antibody. Cholesterol-conjugated antibodies bind to lipid raft domains on the membrane, and because of this enrichment, they show increased antiviral potency. In particular, we find that cholesterol conjugation (i) rescues the antiviral activity of CDRH3-mutated 2F5, (ii) increases the antiviral activity of WT 2F5, (iii) potentiates the non-membrane-binding HIV antibody D5 10–100-fold (depending on the virus strain), and (iv) increases synergy between 2F5 and D5. Conjugation can be made at several positions, including variable and constant domains. Cholesterol conjugation therefore appears to be a general strategy to boost the potency of antiviral antibodies, and, because membrane affinity is engineered outside of the antibody paratope, it can complement affinity maturation strategies. |
| Databáze: | OpenAIRE |
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