HIV-1 neutralization profile and plant-based recombinant expression of actinohivin, an Env glycan-specific lectin devoid of T-cell mitogenic activity
| Autor: | Michelle M. Pickel, Kazunobu Tanno, Nobuyuki Matoba, Charles J. Arntzen, David C. Montefiori, Satoshi Omura, Jason P. Mooney, Haruo Tanaka, Brian W. Barnett, Carl V. Hanson, Huyen Cao, Adam S. Husk, Atsushi Takahashi |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2010 |
| Předmět: |
Sexual transmission
T-Lymphocytes Nicotiana benthamiana lcsh:Medicine Biology Microbiology law.invention Bacterial Proteins Viral envelope Neutralization Tests law Plant virus Microbicide Tobacco Humans Women's Health/Sexually Transmitted Diseases Biotechnology/Plant Biotechnology lcsh:Science Cell Proliferation Virology/Antivirals including Modes of Action and Resistance Syncytium Multidisciplinary lcsh:R Lectin virus diseases biology.organism_classification Virology Recombinant Proteins Virology/Immunodeficiency Viruses HIV-1 Recombinant DNA biology.protein lcsh:Q Mitogens HeLa Cells Research Article |
| Zdroj: | PLoS ONE, Vol 5, Iss 6, p e11143 (2010) PLoS ONE |
| ISSN: | 1932-6203 |
| Popis: | The development of a topical microbicide blocking the sexual transmission of HIV-1 is urgently needed to control the global HIV/AIDS pandemic. The actinomycete-derived lectin actinohivin (AH) is highly specific to a cluster of high-mannose-type glycans uniquely found on the viral envelope (Env). Here, we evaluated AH's candidacy toward a microbicide in terms of in vitro anti-HIV-1 activity, potential side effects, and recombinant producibility. Two validated assay systems based on human peripheral blood mononuclear cell (hPBMC) infection with primary isolates and TZM-bl cell infection with Env-pseudotyped viruses were employed to characterize AH's anti-HIV-1 activity. In hPMBCs, AH exhibited nanomolar neutralizing activity against primary viruses with diverse cellular tropisms, but did not cause mitogenicity or cytotoxicity that are often associated with other anti-HIV lectins. In the TZM-bl-based assay, AH showed broad anti-HIV-1 activity against clinically-relevant, mucosally transmitting strains of clades B and C. By contrast, clade A viruses showed strong resistance to AH. Correlation analysis suggested that HIV-1′s AH susceptibility is significantly linked to the N-glycans at the Env C2 and V4 regions. For recombinant (r)AH expression, we evaluated a tobacco mosaic virus-based system in Nicotiana benthamiana plants as a means to facilitate molecular engineering and cost-effective mass production. Biochemical analysis and an Env-mediated syncytium formation assay demonstrated high-level expression of functional rAH within six days. Taken together, our study revealed AH's cross-clade anti-HIV-1 activity, apparent lack of side effects common to lectins, and robust producibility using plant biotechnology. These findings justify further efforts to develop rAH toward a candidate HIV-1 microbicide. |
| Databáze: | OpenAIRE |
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