Giardia duodenalis extracellular vesicles regulate the proinflammatory immune response in mouse macrophages in vitro via the MAPK, AKT and NF-κB pathways
Abstract Background Giardia duodenalis is an extracellular protozoan parasite that causes giardiasis in mammals. The presentation of giardiasis ranges from asymptomatic to severe diarrhea, and the World Health Organization lists it in the Neglected Diseases Initiative. Extracellular vesicles (EVs) a...
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doaj-dc2695ed0b864676afb845d90f4aa5212021-07-11T11:31:40ZengBMCParasites & Vectors1756-33052021-07-0114111310.1186/s13071-021-04865-5Giardia duodenalis extracellular vesicles regulate the proinflammatory immune response in mouse macrophages in vitro via the MAPK, AKT and NF-κB pathwaysPanpan Zhao0Lili Cao1Xiaocen Wang2Jianhua Li3Jingquan Dong4Nan Zhang5Xin Li6Shan Li7Min Sun8Xichen Zhang9Min Liang10Xudong Pu11Pengtao Gong12Key Laboratory of Zoonosis, College of Veterinary Medicine, Jilin UniversityKey Laboratory of Zoonosis, College of Veterinary Medicine, Jilin UniversityKey Laboratory of Zoonosis, College of Veterinary Medicine, Jilin UniversityKey Laboratory of Zoonosis, College of Veterinary Medicine, Jilin UniversityKey Laboratory of Zoonosis, College of Veterinary Medicine, Jilin UniversityKey Laboratory of Zoonosis, College of Veterinary Medicine, Jilin UniversityKey Laboratory of Zoonosis, College of Veterinary Medicine, Jilin UniversityKey Laboratory of Zoonosis, College of Veterinary Medicine, Jilin UniversityKey Laboratory of Zoonosis, College of Veterinary Medicine, Jilin UniversityKey Laboratory of Zoonosis, College of Veterinary Medicine, Jilin UniversityKey Laboratory of Zoonosis, College of Veterinary Medicine, Jilin UniversityKey Laboratory of Zoonosis, College of Veterinary Medicine, Jilin UniversityKey Laboratory of Zoonosis, College of Veterinary Medicine, Jilin UniversityAbstract Background Giardia duodenalis is an extracellular protozoan parasite that causes giardiasis in mammals. The presentation of giardiasis ranges from asymptomatic to severe diarrhea, and the World Health Organization lists it in the Neglected Diseases Initiative. Extracellular vesicles (EVs) are a key mediator of intracellular communication. Although previous studies have shown that G. intestinalis can regulate a host’s innate immune response, the role of G. intestinalis EVs (GEVs) in triggering a G. intestinalis-induced innate immune response remains to be further explored. Methods In this study, GEVs, G. intestinalis and GEVs + G. intestinalis were inoculated into macrophages, respectively. The transcription and secretion levels of proinflammatory cytokines, including interleukin (IL)-1β, IL-6 and tumor necrosis factor alpha (TNF-α), were measured using real-time quantitative PCR (qPCR) and enzyme-linked immunosorbent assays (ELISAs). The phosphorylation levels of the MAPK, AKT and NF-κB signaling pathways in GEV-stimulated mouse macrophages were examined using western blotting and immunofluorescence methods. The roles of activated pathways in the GEV-triggered inflammatory response were determined using inhibition assays, western blotting and ELISAs. Results The results showed that pretreatment with GEVs enhanced with G. intestinalis (GEVs + G. intestinalis) induced IL-1β, IL-6 and TNF-α transcription and secretion from mouse macrophages compared to stimulation with either GEVs or G. intestinalis alone. Inoculation of mouse macrophages with GEVs upregulated the phosphorylation levels of the p38 MAPK, p44/42 MAPK (Erk1/2), AKT and NF-κB signaling pathways and led to the nuclear translocation of NF-κB p65. Blocking the activated p38, Erk and NF-κB signaling pathways significantly downregulated the secretion of proinflammatory cytokines, and blocking the activated AKT signaling pathway demonstrated reverse effects. Conclusions The results of this study reveal that GEVs can enhance G. intestinalis-induced inflammatory response levels in mouse macrophages through activation of the p38, ERK and NF-κB signaling pathways. The role of GEVs in regulating host cell immune responses may provide insights into exploring the underlying mechanisms in G. intestinalis–host interactions. Graphical abstracthttps://doi.org/10.1186/s13071-021-04865-5Giardia duodenalisExtracellular vesiclesImmune responseMAPK |
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DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Panpan Zhao Lili Cao Xiaocen Wang Jianhua Li Jingquan Dong Nan Zhang Xin Li Shan Li Min Sun Xichen Zhang Min Liang Xudong Pu Pengtao Gong |
spellingShingle |
Panpan Zhao Lili Cao Xiaocen Wang Jianhua Li Jingquan Dong Nan Zhang Xin Li Shan Li Min Sun Xichen Zhang Min Liang Xudong Pu Pengtao Gong Giardia duodenalis extracellular vesicles regulate the proinflammatory immune response in mouse macrophages in vitro via the MAPK, AKT and NF-κB pathways Parasites & Vectors Giardia duodenalis Extracellular vesicles Immune response MAPK |
author_facet |
Panpan Zhao Lili Cao Xiaocen Wang Jianhua Li Jingquan Dong Nan Zhang Xin Li Shan Li Min Sun Xichen Zhang Min Liang Xudong Pu Pengtao Gong |
author_sort |
Panpan Zhao |
title |
Giardia duodenalis extracellular vesicles regulate the proinflammatory immune response in mouse macrophages in vitro via the MAPK, AKT and NF-κB pathways |
title_short |
Giardia duodenalis extracellular vesicles regulate the proinflammatory immune response in mouse macrophages in vitro via the MAPK, AKT and NF-κB pathways |
title_full |
Giardia duodenalis extracellular vesicles regulate the proinflammatory immune response in mouse macrophages in vitro via the MAPK, AKT and NF-κB pathways |
title_fullStr |
Giardia duodenalis extracellular vesicles regulate the proinflammatory immune response in mouse macrophages in vitro via the MAPK, AKT and NF-κB pathways |
title_full_unstemmed |
Giardia duodenalis extracellular vesicles regulate the proinflammatory immune response in mouse macrophages in vitro via the MAPK, AKT and NF-κB pathways |
title_sort |
giardia duodenalis extracellular vesicles regulate the proinflammatory immune response in mouse macrophages in vitro via the mapk, akt and nf-κb pathways |
publisher |
BMC |
series |
Parasites & Vectors |
issn |
1756-3305 |
publishDate |
2021-07-01 |
description |
Abstract Background Giardia duodenalis is an extracellular protozoan parasite that causes giardiasis in mammals. The presentation of giardiasis ranges from asymptomatic to severe diarrhea, and the World Health Organization lists it in the Neglected Diseases Initiative. Extracellular vesicles (EVs) are a key mediator of intracellular communication. Although previous studies have shown that G. intestinalis can regulate a host’s innate immune response, the role of G. intestinalis EVs (GEVs) in triggering a G. intestinalis-induced innate immune response remains to be further explored. Methods In this study, GEVs, G. intestinalis and GEVs + G. intestinalis were inoculated into macrophages, respectively. The transcription and secretion levels of proinflammatory cytokines, including interleukin (IL)-1β, IL-6 and tumor necrosis factor alpha (TNF-α), were measured using real-time quantitative PCR (qPCR) and enzyme-linked immunosorbent assays (ELISAs). The phosphorylation levels of the MAPK, AKT and NF-κB signaling pathways in GEV-stimulated mouse macrophages were examined using western blotting and immunofluorescence methods. The roles of activated pathways in the GEV-triggered inflammatory response were determined using inhibition assays, western blotting and ELISAs. Results The results showed that pretreatment with GEVs enhanced with G. intestinalis (GEVs + G. intestinalis) induced IL-1β, IL-6 and TNF-α transcription and secretion from mouse macrophages compared to stimulation with either GEVs or G. intestinalis alone. Inoculation of mouse macrophages with GEVs upregulated the phosphorylation levels of the p38 MAPK, p44/42 MAPK (Erk1/2), AKT and NF-κB signaling pathways and led to the nuclear translocation of NF-κB p65. Blocking the activated p38, Erk and NF-κB signaling pathways significantly downregulated the secretion of proinflammatory cytokines, and blocking the activated AKT signaling pathway demonstrated reverse effects. Conclusions The results of this study reveal that GEVs can enhance G. intestinalis-induced inflammatory response levels in mouse macrophages through activation of the p38, ERK and NF-κB signaling pathways. The role of GEVs in regulating host cell immune responses may provide insights into exploring the underlying mechanisms in G. intestinalis–host interactions. Graphical abstract |
topic |
Giardia duodenalis Extracellular vesicles Immune response MAPK |
url |
https://doi.org/10.1186/s13071-021-04865-5 |
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