HIV-1 neutralization profile and plant-based recombinant expression of actinohivin, an Env glycan-specific lectin devoid of T-cell mitogenic activity.

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). Her...

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Main Authors: Nobuyuki Matoba, Adam S Husk, Brian W Barnett, Michelle M Pickel, Charles J Arntzen, David C Montefiori, Atsushi Takahashi, Kazunobu Tanno, Satoshi Omura, Huyen Cao, Jason P Mooney, Carl V Hanson, Haruo Tanaka
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2010-06-01
Series:PLoS ONE
Online Access:https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0011143&type=printable
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author Nobuyuki Matoba
Adam S Husk
Brian W Barnett
Michelle M Pickel
Charles J Arntzen
David C Montefiori
Atsushi Takahashi
Kazunobu Tanno
Satoshi Omura
Huyen Cao
Jason P Mooney
Carl V Hanson
Haruo Tanaka
author_facet Nobuyuki Matoba
Adam S Husk
Brian W Barnett
Michelle M Pickel
Charles J Arntzen
David C Montefiori
Atsushi Takahashi
Kazunobu Tanno
Satoshi Omura
Huyen Cao
Jason P Mooney
Carl V Hanson
Haruo Tanaka
author_sort Nobuyuki Matoba
collection DOAJ
description 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.
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spelling doaj-art-8da529b66fe640bba203fa649dd60c172025-08-20T02:31:48ZengPublic Library of Science (PLoS)PLoS ONE1932-62032010-06-0156e1114310.1371/journal.pone.0011143HIV-1 neutralization profile and plant-based recombinant expression of actinohivin, an Env glycan-specific lectin devoid of T-cell mitogenic activity.Nobuyuki MatobaAdam S HuskBrian W BarnettMichelle M PickelCharles J ArntzenDavid C MontefioriAtsushi TakahashiKazunobu TannoSatoshi OmuraHuyen CaoJason P MooneyCarl V HansonHaruo TanakaThe 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.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0011143&type=printable
spellingShingle Nobuyuki Matoba
Adam S Husk
Brian W Barnett
Michelle M Pickel
Charles J Arntzen
David C Montefiori
Atsushi Takahashi
Kazunobu Tanno
Satoshi Omura
Huyen Cao
Jason P Mooney
Carl V Hanson
Haruo Tanaka
HIV-1 neutralization profile and plant-based recombinant expression of actinohivin, an Env glycan-specific lectin devoid of T-cell mitogenic activity.
PLoS ONE
title HIV-1 neutralization profile and plant-based recombinant expression of actinohivin, an Env glycan-specific lectin devoid of T-cell mitogenic activity.
title_full HIV-1 neutralization profile and plant-based recombinant expression of actinohivin, an Env glycan-specific lectin devoid of T-cell mitogenic activity.
title_fullStr HIV-1 neutralization profile and plant-based recombinant expression of actinohivin, an Env glycan-specific lectin devoid of T-cell mitogenic activity.
title_full_unstemmed HIV-1 neutralization profile and plant-based recombinant expression of actinohivin, an Env glycan-specific lectin devoid of T-cell mitogenic activity.
title_short HIV-1 neutralization profile and plant-based recombinant expression of actinohivin, an Env glycan-specific lectin devoid of T-cell mitogenic activity.
title_sort hiv 1 neutralization profile and plant based recombinant expression of actinohivin an env glycan specific lectin devoid of t cell mitogenic activity
url https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0011143&type=printable
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