Antiadenoviral effects of N-chlorotaurine in vitro confirmed by quantitative polymerase chain reaction methods

Eiichi Uchio1, Hirotoshi Inoue1, Kazuaki Kadonosono21Department of Ophthalmology, Fukuoka University School of Medicine, Fukuoka, Japan; 2Department of Ophthalmology, Yokohama City University Medical Center, Yokohama, JapanPurpose: Adenoviral keratoconjunctivitis is recognized as one of the major pa...

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Autores principales:	Eiichi Uchio, Hirotoshi Inoue, Kazuaki Kadonosono
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2010
Materias:	Ophthalmology RE1-994
Acceso en línea:	https://doaj.org/article/066115e4dca744309b1cdee6c5f93e27
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spelling	oai:doaj.org-article:066115e4dca744309b1cdee6c5f93e272021-12-02T02:58:53ZAntiadenoviral effects of N-chlorotaurine in vitro confirmed by quantitative polymerase chain reaction methods1177-54671177-5483https://doaj.org/article/066115e4dca744309b1cdee6c5f93e272010-11-01T00:00:00Zhttp://www.dovepress.com/antiadenoviral-effects-of-n-chlorotaurine-in-vitro-confirmed-by-quanti-a5696https://doaj.org/toc/1177-5467https://doaj.org/toc/1177-5483Eiichi Uchio1, Hirotoshi Inoue1, Kazuaki Kadonosono21Department of Ophthalmology, Fukuoka University School of Medicine, Fukuoka, Japan; 2Department of Ophthalmology, Yokohama City University Medical Center, Yokohama, JapanPurpose: Adenoviral keratoconjunctivitis is recognized as one of the major pathogens of ophthalmological nosocomial infection worldwide. N-Chlorotaurine (Cl&ndash;HN&ndash;CH2&ndash;CH2&ndash;SO3H, NCT) is the N-chloro derivative of the amino acid taurine, which is an oxidant produced by human granulocytes and monocytes during inflammatory reactions. Using conventional viral plaque assay, it was previously shown that NCT causes inactivation of several human adenovirus (HAdV) serotypes. In this study, we evaluated the antiadenoviral effect of NCT by quantitative polymerase chain reaction (PCR) methods.Methods: A549 cells were used for viral cell culture, and HAdV serotypes 3, 4, 8, 19, and 37 were used. After calculating 50% cytotoxic concentration (CC50) of NCT by MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) method, HAdV was cultured with NCT for 7 days, and extracted adenoviral DNA was quantitatively measured by real-time PCR.Results: A statistically significant (P &lt; 0.05) dose-dependent inhibition was indicated for all serotypes except HAdV type 4 (HAdV4), which was maximally inhibited by only ~50%. Among the serotypes, NCT was particularly effective against HAdV8, HAdV19a, and HAdV37. The 50% effective concentration (EC50) obtained by real-time PCR of NCT ranged between 49 and 256 &micro;M. EC50 of NCT against HAdV3 was slightly higher than that against serotypes of species D. The selective index (CC50/EC50) ranged between 41 and 60 except for HAdV4 (11.5).Conclusions: These results show that NCT has an antiviral effect against most serotypes of human HAdV inducing keratoconjunctivitis, indicating its possible therapeutic use.Keywords: adenovirus, N-chlorotaurine, epidemic keratoconjunctivitis, antiviral agent Eiichi UchioHirotoshi InoueKazuaki KadonosonoDove Medical PressarticleOphthalmologyRE1-994ENClinical Ophthalmology, Vol 2010, Iss default, Pp 1325-1329 (2010)
institution	DOAJ
collection	DOAJ
language	EN
topic	Ophthalmology RE1-994
spellingShingle	Ophthalmology RE1-994 Eiichi Uchio Hirotoshi Inoue Kazuaki Kadonosono Antiadenoviral effects of N-chlorotaurine in vitro confirmed by quantitative polymerase chain reaction methods
description	Eiichi Uchio1, Hirotoshi Inoue1, Kazuaki Kadonosono21Department of Ophthalmology, Fukuoka University School of Medicine, Fukuoka, Japan; 2Department of Ophthalmology, Yokohama City University Medical Center, Yokohama, JapanPurpose: Adenoviral keratoconjunctivitis is recognized as one of the major pathogens of ophthalmological nosocomial infection worldwide. N-Chlorotaurine (Cl&ndash;HN&ndash;CH2&ndash;CH2&ndash;SO3H, NCT) is the N-chloro derivative of the amino acid taurine, which is an oxidant produced by human granulocytes and monocytes during inflammatory reactions. Using conventional viral plaque assay, it was previously shown that NCT causes inactivation of several human adenovirus (HAdV) serotypes. In this study, we evaluated the antiadenoviral effect of NCT by quantitative polymerase chain reaction (PCR) methods.Methods: A549 cells were used for viral cell culture, and HAdV serotypes 3, 4, 8, 19, and 37 were used. After calculating 50% cytotoxic concentration (CC50) of NCT by MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) method, HAdV was cultured with NCT for 7 days, and extracted adenoviral DNA was quantitatively measured by real-time PCR.Results: A statistically significant (P &lt; 0.05) dose-dependent inhibition was indicated for all serotypes except HAdV type 4 (HAdV4), which was maximally inhibited by only ~50%. Among the serotypes, NCT was particularly effective against HAdV8, HAdV19a, and HAdV37. The 50% effective concentration (EC50) obtained by real-time PCR of NCT ranged between 49 and 256 &micro;M. EC50 of NCT against HAdV3 was slightly higher than that against serotypes of species D. The selective index (CC50/EC50) ranged between 41 and 60 except for HAdV4 (11.5).Conclusions: These results show that NCT has an antiviral effect against most serotypes of human HAdV inducing keratoconjunctivitis, indicating its possible therapeutic use.Keywords: adenovirus, N-chlorotaurine, epidemic keratoconjunctivitis, antiviral agent
format	article
author	Eiichi Uchio Hirotoshi Inoue Kazuaki Kadonosono
author_facet	Eiichi Uchio Hirotoshi Inoue Kazuaki Kadonosono
author_sort	Eiichi Uchio
title	Antiadenoviral effects of N-chlorotaurine in vitro confirmed by quantitative polymerase chain reaction methods
title_short	Antiadenoviral effects of N-chlorotaurine in vitro confirmed by quantitative polymerase chain reaction methods
title_full	Antiadenoviral effects of N-chlorotaurine in vitro confirmed by quantitative polymerase chain reaction methods
title_fullStr	Antiadenoviral effects of N-chlorotaurine in vitro confirmed by quantitative polymerase chain reaction methods
title_full_unstemmed	Antiadenoviral effects of N-chlorotaurine in vitro confirmed by quantitative polymerase chain reaction methods
title_sort	antiadenoviral effects of n-chlorotaurine in vitro confirmed by quantitative polymerase chain reaction methods
publisher	Dove Medical Press
publishDate	2010
url	https://doaj.org/article/066115e4dca744309b1cdee6c5f93e27
work_keys_str_mv	AT eiichiuchio antiadenoviraleffectsofnchlorotaurineinvitroconfirmedbyquantitativepolymerasechainreactionmethods AT hirotoshiinoue antiadenoviraleffectsofnchlorotaurineinvitroconfirmedbyquantitativepolymerasechainreactionmethods AT kazuakikadonosono antiadenoviraleffectsofnchlorotaurineinvitroconfirmedbyquantitativepolymerasechainreactionmethods
_version_	1718402021217796096

Antiadenoviral effects of N-chlorotaurine in vitro confirmed by quantitative polymerase chain reaction methods

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