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Giardia VSPAS7 protein attenuates Giardia intestinalis-induced host macrophage pyroptosis

BACKGROUND: The unicellular protozoan parasite Giardia intestinalis, which primarily infects humans and animals such as cattle and sheep, is having a major negative impact on public health. Giardia is able to evade the recognition and elimination of the host immune system because of the trophozoite...

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Autores principales: Sun, Min, Zhao, Zhiteng, Li, Ying, Cao, Lili, Li, Jianhua, Zhang, Xichen, Li, Xin, Zhang, Nan, Cheng, Shuqin, Wang, Xiaocen, Gong, Pengtao
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10566177/
https://www.ncbi.nlm.nih.gov/pubmed/37821972
http://dx.doi.org/10.1186/s13071-023-05949-0
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author Sun, Min
Zhao, Zhiteng
Li, Ying
Cao, Lili
Li, Jianhua
Zhang, Xichen
Li, Xin
Zhang, Nan
Cheng, Shuqin
Wang, Xiaocen
Gong, Pengtao
author_facet Sun, Min
Zhao, Zhiteng
Li, Ying
Cao, Lili
Li, Jianhua
Zhang, Xichen
Li, Xin
Zhang, Nan
Cheng, Shuqin
Wang, Xiaocen
Gong, Pengtao
author_sort Sun, Min
collection PubMed
description BACKGROUND: The unicellular protozoan parasite Giardia intestinalis, which primarily infects humans and animals such as cattle and sheep, is having a major negative impact on public health. Giardia is able to evade the recognition and elimination of the host immune system because of the trophozoite surface and extracellular vesicles (EVs) covered by variant-specific surface proteins (VSPs). As key proteins for immune evasion, whether VSPs can regulate Giardia-induced pyroptosis and promote Giardia evasion of host immune responses has not been reported. METHODS: To examine the role of Giardia VSPAS7 on Giardia-induced activation of the signaling pathway, secretion of pro-inflammatory cytokines, pyroptosis and the mechanism involved, we constructed the pcDNA3.1-vspas7 expression plasmid and transfected this plasmid into mouse macrophages. Key proteins for pyroptosis, IL-1β secretion and LDH release were detected in pcDNA3.1-vspas7-transfected wild-type (WT) cells and NLRP3-deficient cells by western blot, ELISA and LDH assays, respectively. The interactions of Giardia VSPAS7 and mouse NLRP3 were examined using immunofluorescence assays (IFA), co-immunoprecipitation (Co-IP) and bimolecular fluorescence complementation (BiFC) assays. RESULTS: VSPAS7 could decrease the levels of phosphorylated-p65 (P-p65), P-IκBα and P-ERK caused by Giardia and reduce the production levels of Giardia-induced pro-inflammatory cytokine IL-6, IL-12 p40 and TNF-α. The results showed that VSPAS7 inhibited Giardia-mediated activation of NF-κB, ERK/MAPK signaling and secretion of pro-inflammatory cytokines. Furthermore, VSPAS7 suppressed Giardia-induced macrophage pyroptosis by reducing GSDMD cleavage, caspase-1 activation, IL-1β secretion and LDH release. We further found that VSPAS7 could interact with mouse NLRP3 directly, and in NLRP3-deficient cells the suppression of Giardia-induced macrophage pyroptosis by VSPAS7 was significantly attenuated. CONCLUSIONS: Overall, VSPAS7 could inhibit Giardia-induced activation of signaling pathways and pyroptosis in host macrophages, allowing Giardia evasion of host immune responses. Studies on Giardia VSP-mediated immune evasion provide an important theoretical basis for in-depth studies on Giardia pathogenicity. GRAPHICAL ABSTRACT: [Image: see text] SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13071-023-05949-0.
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spelling pubmed-105661772023-10-12 Giardia VSPAS7 protein attenuates Giardia intestinalis-induced host macrophage pyroptosis Sun, Min Zhao, Zhiteng Li, Ying Cao, Lili Li, Jianhua Zhang, Xichen Li, Xin Zhang, Nan Cheng, Shuqin Wang, Xiaocen Gong, Pengtao Parasit Vectors Research BACKGROUND: The unicellular protozoan parasite Giardia intestinalis, which primarily infects humans and animals such as cattle and sheep, is having a major negative impact on public health. Giardia is able to evade the recognition and elimination of the host immune system because of the trophozoite surface and extracellular vesicles (EVs) covered by variant-specific surface proteins (VSPs). As key proteins for immune evasion, whether VSPs can regulate Giardia-induced pyroptosis and promote Giardia evasion of host immune responses has not been reported. METHODS: To examine the role of Giardia VSPAS7 on Giardia-induced activation of the signaling pathway, secretion of pro-inflammatory cytokines, pyroptosis and the mechanism involved, we constructed the pcDNA3.1-vspas7 expression plasmid and transfected this plasmid into mouse macrophages. Key proteins for pyroptosis, IL-1β secretion and LDH release were detected in pcDNA3.1-vspas7-transfected wild-type (WT) cells and NLRP3-deficient cells by western blot, ELISA and LDH assays, respectively. The interactions of Giardia VSPAS7 and mouse NLRP3 were examined using immunofluorescence assays (IFA), co-immunoprecipitation (Co-IP) and bimolecular fluorescence complementation (BiFC) assays. RESULTS: VSPAS7 could decrease the levels of phosphorylated-p65 (P-p65), P-IκBα and P-ERK caused by Giardia and reduce the production levels of Giardia-induced pro-inflammatory cytokine IL-6, IL-12 p40 and TNF-α. The results showed that VSPAS7 inhibited Giardia-mediated activation of NF-κB, ERK/MAPK signaling and secretion of pro-inflammatory cytokines. Furthermore, VSPAS7 suppressed Giardia-induced macrophage pyroptosis by reducing GSDMD cleavage, caspase-1 activation, IL-1β secretion and LDH release. We further found that VSPAS7 could interact with mouse NLRP3 directly, and in NLRP3-deficient cells the suppression of Giardia-induced macrophage pyroptosis by VSPAS7 was significantly attenuated. CONCLUSIONS: Overall, VSPAS7 could inhibit Giardia-induced activation of signaling pathways and pyroptosis in host macrophages, allowing Giardia evasion of host immune responses. Studies on Giardia VSP-mediated immune evasion provide an important theoretical basis for in-depth studies on Giardia pathogenicity. GRAPHICAL ABSTRACT: [Image: see text] SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13071-023-05949-0. BioMed Central 2023-10-11 /pmc/articles/PMC10566177/ /pubmed/37821972 http://dx.doi.org/10.1186/s13071-023-05949-0 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This 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/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://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
Sun, Min
Zhao, Zhiteng
Li, Ying
Cao, Lili
Li, Jianhua
Zhang, Xichen
Li, Xin
Zhang, Nan
Cheng, Shuqin
Wang, Xiaocen
Gong, Pengtao
Giardia VSPAS7 protein attenuates Giardia intestinalis-induced host macrophage pyroptosis
title Giardia VSPAS7 protein attenuates Giardia intestinalis-induced host macrophage pyroptosis
title_full Giardia VSPAS7 protein attenuates Giardia intestinalis-induced host macrophage pyroptosis
title_fullStr Giardia VSPAS7 protein attenuates Giardia intestinalis-induced host macrophage pyroptosis
title_full_unstemmed Giardia VSPAS7 protein attenuates Giardia intestinalis-induced host macrophage pyroptosis
title_short Giardia VSPAS7 protein attenuates Giardia intestinalis-induced host macrophage pyroptosis
title_sort giardia vspas7 protein attenuates giardia intestinalis-induced host macrophage pyroptosis
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10566177/
https://www.ncbi.nlm.nih.gov/pubmed/37821972
http://dx.doi.org/10.1186/s13071-023-05949-0
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