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TgROP18 targets IL20RB for host-defense-related-STAT3 activation during Toxoplasma gondii infection

BACKGROUND: Toxoplasma gondii is an opportunistic protozoan infecting almost one-third of the world’s population. Toxoplasma gondii rhoptry protein 18 (TgROP18) is a key virulence factor determining the parasite’s acute virulence and is secreted into host cells during infection. We previously identi...

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Autores principales: Kong, Ling, Jiang, Dan, He, Cheng, Xia, Jing, Wei, Haixia, Zhou, Lijuan, Peng, Hongjuan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7412674/
https://www.ncbi.nlm.nih.gov/pubmed/32767999
http://dx.doi.org/10.1186/s13071-020-04251-7
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author Kong, Ling
Jiang, Dan
He, Cheng
Xia, Jing
Wei, Haixia
Zhou, Lijuan
Peng, Hongjuan
author_facet Kong, Ling
Jiang, Dan
He, Cheng
Xia, Jing
Wei, Haixia
Zhou, Lijuan
Peng, Hongjuan
author_sort Kong, Ling
collection PubMed
description BACKGROUND: Toxoplasma gondii is an opportunistic protozoan infecting almost one-third of the world’s population. Toxoplasma gondii rhoptry protein 18 (TgROP18) is a key virulence factor determining the parasite’s acute virulence and is secreted into host cells during infection. We previously identified the interaction of TgROP18 and host cell immune-related receptor protein IL20RB, and observed the activation of STAT3 in human keratinocytes (HaCaT) cells infected by the rop16 knockout RH strain, though TgROP16 is regarded as being responsible for host STAT3 activation during T. gondii invasion. Therefore, we hypothesize TgROP18 can activate host STAT3 through binding to IL20RB. METHODS: CRISPR-CAS9 technology was used to generate the ROP16 and ROP18 double knockout RH strain, RH-∆rop16∆rop18. SDS-PAGE and western blot were used to detect STAT3 activation in different HaCaT cells with high endogenous IL20RB expression treated with T. gondii tachyzoites infection, recombinant ROP18, or IL-20. FRET and co-immunoprecipitation (Co-IP) was used to detect the protein-protein interaction. RESULTS: We observed that TgROP18 was involved in a synergic activation of the host JAK/STAT3 pathway together with TgROP16 in human HaCaT cells infected with T. gondii or treated with recombinant TgROP18 protein, stimulating host proinflammatory immune responses such as expression of TNF-α. The effect of recombinant ROP18 on STAT3 phosphorylation was presented in a dose-dependent manner. Additionally, TgROP18 was identified to target IL20RB on its extracellular domain. When we treated different cell lines with the recombinant ROP18, STAT3 phosphorylation could only be observed in the cells with endogenous IL20RB expression, such as HaCaT cells. CONCLUSIONS: These findings indicate that TgROP18-IL20RB interaction upon T. gondii invasion was involved in STAT3 activation, which is associated with host cell defense. [Image: see text]
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spelling pubmed-74126742020-08-10 TgROP18 targets IL20RB for host-defense-related-STAT3 activation during Toxoplasma gondii infection Kong, Ling Jiang, Dan He, Cheng Xia, Jing Wei, Haixia Zhou, Lijuan Peng, Hongjuan Parasit Vectors Research BACKGROUND: Toxoplasma gondii is an opportunistic protozoan infecting almost one-third of the world’s population. Toxoplasma gondii rhoptry protein 18 (TgROP18) is a key virulence factor determining the parasite’s acute virulence and is secreted into host cells during infection. We previously identified the interaction of TgROP18 and host cell immune-related receptor protein IL20RB, and observed the activation of STAT3 in human keratinocytes (HaCaT) cells infected by the rop16 knockout RH strain, though TgROP16 is regarded as being responsible for host STAT3 activation during T. gondii invasion. Therefore, we hypothesize TgROP18 can activate host STAT3 through binding to IL20RB. METHODS: CRISPR-CAS9 technology was used to generate the ROP16 and ROP18 double knockout RH strain, RH-∆rop16∆rop18. SDS-PAGE and western blot were used to detect STAT3 activation in different HaCaT cells with high endogenous IL20RB expression treated with T. gondii tachyzoites infection, recombinant ROP18, or IL-20. FRET and co-immunoprecipitation (Co-IP) was used to detect the protein-protein interaction. RESULTS: We observed that TgROP18 was involved in a synergic activation of the host JAK/STAT3 pathway together with TgROP16 in human HaCaT cells infected with T. gondii or treated with recombinant TgROP18 protein, stimulating host proinflammatory immune responses such as expression of TNF-α. The effect of recombinant ROP18 on STAT3 phosphorylation was presented in a dose-dependent manner. Additionally, TgROP18 was identified to target IL20RB on its extracellular domain. When we treated different cell lines with the recombinant ROP18, STAT3 phosphorylation could only be observed in the cells with endogenous IL20RB expression, such as HaCaT cells. CONCLUSIONS: These findings indicate that TgROP18-IL20RB interaction upon T. gondii invasion was involved in STAT3 activation, which is associated with host cell defense. [Image: see text] BioMed Central 2020-08-07 /pmc/articles/PMC7412674/ /pubmed/32767999 http://dx.doi.org/10.1186/s13071-020-04251-7 Text en © The Author(s) 2020 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Kong, Ling
Jiang, Dan
He, Cheng
Xia, Jing
Wei, Haixia
Zhou, Lijuan
Peng, Hongjuan
TgROP18 targets IL20RB for host-defense-related-STAT3 activation during Toxoplasma gondii infection
title TgROP18 targets IL20RB for host-defense-related-STAT3 activation during Toxoplasma gondii infection
title_full TgROP18 targets IL20RB for host-defense-related-STAT3 activation during Toxoplasma gondii infection
title_fullStr TgROP18 targets IL20RB for host-defense-related-STAT3 activation during Toxoplasma gondii infection
title_full_unstemmed TgROP18 targets IL20RB for host-defense-related-STAT3 activation during Toxoplasma gondii infection
title_short TgROP18 targets IL20RB for host-defense-related-STAT3 activation during Toxoplasma gondii infection
title_sort tgrop18 targets il20rb for host-defense-related-stat3 activation during toxoplasma gondii infection
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7412674/
https://www.ncbi.nlm.nih.gov/pubmed/32767999
http://dx.doi.org/10.1186/s13071-020-04251-7
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