Pub Date : 2025-02-08DOI: 10.1016/j.molimm.2025.01.016
Wei Yan , Hengchao Zhang , Jiashen Zhang , Yaxuan Zhao , Yunhua Wu , Xiaolin Ma , Xiying Luan
Graft-versus-host disease (GVHD) constitutes a severe complication that occurs after allogeneic hematopoietic stem cell transplantation (allo-HSCT), significantly reducing the survival rate of patients. Mesenchymal stem cells (MSCs) are capable of ameliorating the tissue damage caused by GVHD through exerting immunosuppressive effects; however, the relevant mechanisms require further investigation. This study used a GVHD mouse model to explore the therapeutic effects and mechanisms of human placental mesenchymal stem cells (hPMSCs) in mitigating GVHD-induced liver injury. The findings indicated that hPMSCs reduced the proportion of CD8+PD-1+ T cells in both the liver and spleen of GVHD mice, decreased reactive oxygen species (ROS) levels, and upregulated glutathione S transferase (GST) and glutathione (GSH) levels. Consistently, this led to a decrease in the expression of liver fibrosis markers, including alpha-smooth muscle actin (α-SMA) and fibronectin (FN). Moreover, CD8+PD-1+ T cells and ROS were positively correlated with α-SMA and FN, respectively, whereas GST and GSH were negatively correlated with them. hPMSCs with low expression in CD73 attenuated this effect. In vitro studies demonstrated that hPMSCs upregulated the expression of nuclear factor-E2-related factor 2 (Nrf2) via the CD73/adenosine (ADO) pathway, regulated oxidative metabolism, and reduced the number of CD8+PD-1+ T cells. The results suggested that hPMSCs contributed to the regulation of redox homeostasis and reduced the proportion of CD8+PD-1+ T cells through the CD73/ADO/Nrf2 signaling pathway, thereby alleviating liver injury associated with GVHD.
{"title":"Human placental mesenchymal stem cells regulate the antioxidant capacity of CD8+PD-1+ T cells through the CD73/ADO/Nrf2 pathway to protect against liver damage in mice with acute graft-versus-host disease","authors":"Wei Yan , Hengchao Zhang , Jiashen Zhang , Yaxuan Zhao , Yunhua Wu , Xiaolin Ma , Xiying Luan","doi":"10.1016/j.molimm.2025.01.016","DOIUrl":"10.1016/j.molimm.2025.01.016","url":null,"abstract":"<div><div>Graft-versus-host disease (GVHD) constitutes a severe complication that occurs after allogeneic hematopoietic stem cell transplantation (allo-HSCT), significantly reducing the survival rate of patients. Mesenchymal stem cells (MSCs) are capable of ameliorating the tissue damage caused by GVHD through exerting immunosuppressive effects; however, the relevant mechanisms require further investigation. This study used a GVHD mouse model to explore the therapeutic effects and mechanisms of human placental mesenchymal stem cells (hPMSCs) in mitigating GVHD-induced liver injury. The findings indicated that hPMSCs reduced the proportion of CD8<sup>+</sup>PD-1<sup>+</sup> T cells in both the liver and spleen of GVHD mice, decreased reactive oxygen species (ROS) levels, and upregulated glutathione S transferase (GST) and glutathione (GSH) levels. Consistently, this led to a decrease in the expression of liver fibrosis markers, including alpha-smooth muscle actin (α-SMA) and fibronectin (FN). Moreover, CD8<sup>+</sup>PD-1<sup>+</sup> T cells and ROS were positively correlated with α-SMA and FN, respectively, whereas GST and GSH were negatively correlated with them. hPMSCs with low expression in CD73 attenuated this effect. <em>In vitro</em> studies demonstrated that hPMSCs upregulated the expression of nuclear factor-E2-related factor 2 (Nrf2) via the CD73/adenosine (ADO) pathway, regulated oxidative metabolism, and reduced the number of CD8<sup>+</sup>PD-1<sup>+</sup> T cells. The results suggested that hPMSCs contributed to the regulation of redox homeostasis and reduced the proportion of CD8<sup>+</sup>PD-1<sup>+</sup> T cells through the CD73/ADO/Nrf2 signaling pathway, thereby alleviating liver injury associated with GVHD.</div></div>","PeriodicalId":18938,"journal":{"name":"Molecular immunology","volume":"179 ","pages":"Pages 71-83"},"PeriodicalIF":3.2,"publicationDate":"2025-02-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143350883","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Sepsis, a common clinical complication, leads to multi-organ damage due to systemic infection and currently lacks effective therapeutic drugs. Polysaccharide derived from Tetrastigma hemsleyanum Diels et Gilg (TH), abbreviated as THP, is a water-soluble component extracted from TH, exhibiting anti-inflammatory and immunomodulatory properties. This study aims to investigate the effects and mechanisms of THP in sepsis. Results demonstrated that THP reduced neutrophils in the peripheral blood of mice established by cecal ligation and puncture (CLP), and inhibited IL-6 and MCP-1 in plasma, thereby improving systemic inflammation. THP ameliorated pulmonary edema, mitigated lung and liver histopathological injuries, reduced infiltration of neutrophils and macrophages in the lung and liver, and inhibited the TNF-α, IL-6, IL-1β, and MCP-1 transcription in both organs. Additionally, THP decreased myeloid cells, neutrophils, monocytes, and Tregs in the spleens of septic mice, while increasing T cells, CD4+ T cells, and CD8+ T cells, thereby restoring immune imbalance. Mechanistically, THP attenuated sepsis by inhibiting the overactivation of the TLR4/NF-κB/COX-2 pathway, and reducing PD-1, PD-L1, IDO1 in the lung, and PD-1, PD-L1 in the liver of septic mice. In conclusion, this study provides theoretical support for the potential application of THP in sepsis treatment.
{"title":"Alleviation of sepsis-induced lung and liver injury by polysaccharides from Tetrastigma hemsleyanum Diels et Gilg via suppression of TLR4/NF-κB/COX-2 pathway and modulation of immune checkpoint molecules","authors":"Mengjia Zhao , Senmiao Chen , Jingwen Xu , Fangmei Zhou , Mingyuan Zhou , Shasha Tian , Zhishan Ding , Yuchi Chen","doi":"10.1016/j.molimm.2025.02.002","DOIUrl":"10.1016/j.molimm.2025.02.002","url":null,"abstract":"<div><div>Sepsis, a common clinical complication, leads to multi-organ damage due to systemic infection and currently lacks effective therapeutic drugs. Polysaccharide derived from <em>Tetrastigma hemsleyanum</em> Diels et Gilg (TH), abbreviated as THP, is a water-soluble component extracted from TH, exhibiting anti-inflammatory and immunomodulatory properties. This study aims to investigate the effects and mechanisms of THP in sepsis. Results demonstrated that THP reduced neutrophils in the peripheral blood of mice established by cecal ligation and puncture (CLP), and inhibited IL-6 and MCP-1 in plasma, thereby improving systemic inflammation. THP ameliorated pulmonary edema, mitigated lung and liver histopathological injuries, reduced infiltration of neutrophils and macrophages in the lung and liver, and inhibited the TNF-α, IL-6, IL-1β, and MCP-1 transcription in both organs. Additionally, THP decreased myeloid cells, neutrophils, monocytes, and Tregs in the spleens of septic mice, while increasing T cells, CD4<sup>+</sup> T cells, and CD8<sup>+</sup> T cells, thereby restoring immune imbalance. Mechanistically, THP attenuated sepsis by inhibiting the overactivation of the TLR4/NF-κB/COX-2 pathway, and reducing PD-1, PD-L1, IDO1 in the lung, and PD-1, PD-L1 in the liver of septic mice. In conclusion, this study provides theoretical support for the potential application of THP in sepsis treatment.</div></div>","PeriodicalId":18938,"journal":{"name":"Molecular immunology","volume":"179 ","pages":"Pages 52-64"},"PeriodicalIF":3.2,"publicationDate":"2025-02-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143316437","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
There is ample evidence that ubiquitin-conjugating enzyme E2I (UBE2I) is involved in progression of diverse cancers. However, the influence of UBE2I on ovarian cancer (OC) has been poorly reported. This study tries to discover the mechanisms and functions of UBE2I in OC. Relative mRNA expression of UBE2I, CD86, iNOS, MHC II and programmed death ligand 1 (PD-L1) was detected through qRT-PCR. We identified UBE2I, Vimentin, E-Cadherin, N-Cadherin and Ki67 protein expression levels in tumor tissues through immunohistochemistry staining. Protein levels of UBE2I, cleaved caspase-3, cleaved PARP, E-cadherin, N-cadherin and Vimentin were detected through western blot. Cell viability, invasion, and migration were examined by means of cell counting kit-8 (CCK-8), transwell, and wound healing assays. Immunofluorescence was used to detect colocalization between UBE2I and CD68. We assessed expression levels of IFN-γ and TNF-α via flow cytometry and ELISA. We used the TUNEL assay to assess tumor cell apoptosis. Glycolysis was assessed through the consumption of glucose, ATP production, production of lactate, and extracellular acidification rate. For establishing a xenograft model, OC cells were subcutaneously injected into mice. UBE2I expression was boosted in OC cells and tissues, which was negatively associated with OC patients’ prognosis. Silencing of UBE2I suppressed OC cell proliferation, invasion, EMT (epithelial-to-mesenchymal transition) and migration. UBE2I inhibition promoted macrophages toward the M1 phenotype and macrophage viability. After deletion of UBE2I in vivo, mice tumor growth and EMT were suppressed, and apoptosis of tumor cells was increased. Meantime, an increasing proportion of CD86+ TAMs (tumor-associated macrophages) was observed after the deletion of UBE2I. Besides, increases in consumption of glucose, lactate production, ATP production and ECAR in THP-1 cells were observed by silencing of UBE2I; however, glycolysis inhibitor reversed UBE2I-mediated polarization of M1 macrophages in a dose-dependent fashion. Importantly, UBE2I-mediated M1 macrophages promoted PD-L1 expression. Furthermore, the combinatorial therapy of UBE2I inhibitor plus anti-PD-1 repressed tumor growth, reduced Ki67 expression, and promoted apoptosis in tumor cells, exhibiting higher efficiency than UBE2I inhibitor/anti-PD-L1 alone. UBE2I inhibition regulated polarization of M1 macrophages via glycolysis and improved anti-PD-L1 immunotherapy efficacy, paving a novel avenue to prevent OC development.
{"title":"UBE2I depletion regulated tumor-associated macrophage polarization into M1 type through reprogramming glycolysis and increases immunotherapy efficacy of anti-PD-L1 in ovarian cancer","authors":"Lei Zhao , Yuxin Zhang , Jinming Wang, Dongliang Li, Xuewei Hao","doi":"10.1016/j.molimm.2025.01.007","DOIUrl":"10.1016/j.molimm.2025.01.007","url":null,"abstract":"<div><div>There is ample evidence that ubiquitin-conjugating enzyme E2I (UBE2I) is involved in progression of diverse cancers. However, the influence of UBE2I on ovarian cancer (OC) has been poorly reported. This study tries to discover the mechanisms and functions of UBE2I in OC. Relative mRNA expression of UBE2I, CD86, iNOS, MHC II and programmed death ligand 1 (PD-L1) was detected through qRT-PCR. We identified UBE2I, Vimentin, E-Cadherin, N-Cadherin and Ki67 protein expression levels in tumor tissues through immunohistochemistry staining. Protein levels of UBE2I, cleaved caspase-3, cleaved PARP, E-cadherin, N-cadherin and Vimentin were detected through western blot. Cell viability, invasion, and migration were examined by means of cell counting kit-8 (CCK-8), transwell, and wound healing assays. Immunofluorescence was used to detect colocalization between UBE2I and CD68. We assessed expression levels of IFN-γ and TNF-α via flow cytometry and ELISA. We used the TUNEL assay to assess tumor cell apoptosis. Glycolysis was assessed through the consumption of glucose, ATP production, production of lactate, and extracellular acidification rate. For establishing a xenograft model, OC cells were subcutaneously injected into mice. UBE2I expression was boosted in OC cells and tissues, which was negatively associated with OC patients’ prognosis. Silencing of UBE2I suppressed OC cell proliferation, invasion, EMT (epithelial-to-mesenchymal transition) and migration. UBE2I inhibition promoted macrophages toward the M1 phenotype and macrophage viability. After deletion of UBE2I <em>in vivo</em>, mice tumor growth and EMT were suppressed, and apoptosis of tumor cells was increased. Meantime, an increasing proportion of CD86+ TAMs (tumor-associated macrophages) was observed after the deletion of UBE2I. Besides, increases in consumption of glucose, lactate production, ATP production and ECAR in THP-1 cells were observed by silencing of UBE2I; however, glycolysis inhibitor reversed UBE2I-mediated polarization of M1 macrophages in a dose-dependent fashion. Importantly, UBE2I-mediated M1 macrophages promoted PD-L1 expression. Furthermore, the combinatorial therapy of UBE2I inhibitor plus anti-PD-1 repressed tumor growth, reduced Ki67 expression, and promoted apoptosis in tumor cells, exhibiting higher efficiency than UBE2I inhibitor/anti-PD-L1 alone. UBE2I inhibition regulated polarization of M1 macrophages via glycolysis and improved anti-PD-L1 immunotherapy efficacy, paving a novel avenue to prevent OC development.</div></div>","PeriodicalId":18938,"journal":{"name":"Molecular immunology","volume":"179 ","pages":"Pages 29-41"},"PeriodicalIF":3.2,"publicationDate":"2025-02-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143316438","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-06DOI: 10.1016/j.molimm.2025.01.015
Xinran Wang , Huazhong Xiong , Juhui Qiao , Wanying Zhang , Siming Wang , Meichen Liu , Shichao Liu
To examine the protective effect and mechanism of the original extract of Dendrobium officinale (DE) on gastric lesions caused by aspirin (ASA) facilitate the development of traditional Chinese medicine products. In this study, a gastric lesions rat model was established, and then the rats were treated with DE for 5 days. We found that ASA-induced gastric mucosal detachment and hemorrhagic lesions in rats improved after DE supplementation. Meanwhile, effectively inhibits the secretion of inflammatory mediators (IL-6, IL-1β, TNF-α, MPO, COX-2) and upregulates the activity of antioxidant core components (T-SOD, GSH-Px) and defense factors (TFFs, EGF, EGFR) in gastric tissues. It was further found that DE exerted a defensive effect on the gastric mucosa in association with NF-κB/Nrf-2/HO-1 signaling. In conclusion, DE protects the gastric mucosa from oxidative stress, improves its defenses, as well as being a potential gastric mucosal nutrient in the future, preventing ASA-induced gastric lesions.
{"title":"Protective effect of Dendrobium officinale extract on aspirin-induced gastric lesions in rats","authors":"Xinran Wang , Huazhong Xiong , Juhui Qiao , Wanying Zhang , Siming Wang , Meichen Liu , Shichao Liu","doi":"10.1016/j.molimm.2025.01.015","DOIUrl":"10.1016/j.molimm.2025.01.015","url":null,"abstract":"<div><div>To examine the protective effect and mechanism of the original extract of <em>Dendrobium officinale</em> (DE) on gastric lesions caused by aspirin (ASA) facilitate the development of traditional Chinese medicine products. In this study, a gastric lesions rat model was established, and then the rats were treated with DE for 5 days. We found that ASA-induced gastric mucosal detachment and hemorrhagic lesions in rats improved after DE supplementation. Meanwhile, effectively inhibits the secretion of inflammatory mediators (IL-6, IL-1β, TNF-α, MPO, COX-2) and upregulates the activity of antioxidant core components (T-SOD, GSH-Px) and defense factors (TFFs, EGF, EGFR) in gastric tissues. It was further found that DE exerted a defensive effect on the gastric mucosa in association with NF-κB/Nrf-2/HO-1 signaling. In conclusion, DE protects the gastric mucosa from oxidative stress, improves its defenses, as well as being a potential gastric mucosal nutrient in the future, preventing ASA-induced gastric lesions.</div></div>","PeriodicalId":18938,"journal":{"name":"Molecular immunology","volume":"179 ","pages":"Pages 42-51"},"PeriodicalIF":3.2,"publicationDate":"2025-02-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143316436","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-05DOI: 10.1016/j.molimm.2025.01.002
Ke Yan , Jiarong Bian , Liang He , Bingwei Song , Linhai Shen , Yong Zhen
Objective
We aimed to explore the role of SIRT3 in ameliorating neuroinflammation caused by hypoxia-ischemia (HI).
Methods
A rat model of HI was established, and 48 hours prior to constructing the HI model, the rats received an intracerebroventricular injection of a recombinant adeno-associated virus type 9 vector. TTC and Nissl staining assessed the effects of SIRT3 on cerebral infarction and brain atrophy in HI rats. Neuroinflammation was evaluated by investigating IL-1β and MPO positive cells, and ELISA for determining inflammatory cytokines. IBA-1, CD68, and iNOS positive microglia and NLRP3 activation-related protein expression were also detected. SIRT3 was overexpressed in oxygen glucose deprivation (OGD)-induced microglia model, where cell morphology and expressions of pro-inflammatory cytokines and NLRP3 inflammasome activation-related proteins were examined. Additionally, neurons co-cultured with SIRT3-overexpressing microglia were analyzed for mitochondrial damage and apoptosis.
Results
SIRT3 alleviated cerebral infarction and atrophy in HI rats. It also inhibited neuroinflammation, reducing IL-1β and MPO positive cells, and lowered the levels of pro-inflammatory cytokines. In both HI rat model and OGD cell model, SIRT3 inhibited excessive activation of microglia and NLRP3 inflammasome. Furthermore, co-culturing neurons with SIRT3-overexpressing microglia resulted in reduced neuronal apoptosis and improved mitochondrial function, evidenced by lower ROS levels, alleviated mitochondrial depolarization and increased ATP production.
Conclusion
SIRT3 restrains pro-inflammatory microglia and NLRP3 inflammasome and alleviates neuroinflammation following HI brain injury.
{"title":"SIRT3 mitigates neuroinflammation and mitochondrial damage post-hypoxic-ischemic brain injury","authors":"Ke Yan , Jiarong Bian , Liang He , Bingwei Song , Linhai Shen , Yong Zhen","doi":"10.1016/j.molimm.2025.01.002","DOIUrl":"10.1016/j.molimm.2025.01.002","url":null,"abstract":"<div><h3>Objective</h3><div>We aimed to explore the role of SIRT3 in ameliorating neuroinflammation caused by hypoxia-ischemia (HI).</div></div><div><h3>Methods</h3><div>A rat model of HI was established, and 48 hours prior to constructing the HI model, the rats received an intracerebroventricular injection of a recombinant adeno-associated virus type 9 vector. TTC and Nissl staining assessed the effects of SIRT3 on cerebral infarction and brain atrophy in HI rats. Neuroinflammation was evaluated by investigating IL-1β and MPO positive cells, and ELISA for determining inflammatory cytokines. IBA-1, CD68, and iNOS positive microglia and NLRP3 activation-related protein expression were also detected. SIRT3 was overexpressed in oxygen glucose deprivation (OGD)-induced microglia model, where cell morphology and expressions of pro-inflammatory cytokines and NLRP3 inflammasome activation-related proteins were examined. Additionally, neurons co-cultured with SIRT3-overexpressing microglia were analyzed for mitochondrial damage and apoptosis.</div></div><div><h3>Results</h3><div>SIRT3 alleviated cerebral infarction and atrophy in HI rats. It also inhibited neuroinflammation, reducing IL-1β and MPO positive cells, and lowered the levels of pro-inflammatory cytokines. In both HI rat model and OGD cell model, SIRT3 inhibited excessive activation of microglia and NLRP3 inflammasome. Furthermore, co-culturing neurons with SIRT3-overexpressing microglia resulted in reduced neuronal apoptosis and improved mitochondrial function, evidenced by lower ROS levels, alleviated mitochondrial depolarization and increased ATP production.</div></div><div><h3>Conclusion</h3><div>SIRT3 restrains pro-inflammatory microglia and NLRP3 inflammasome and alleviates neuroinflammation following HI brain injury.</div></div>","PeriodicalId":18938,"journal":{"name":"Molecular immunology","volume":"179 ","pages":"Pages 18-28"},"PeriodicalIF":3.2,"publicationDate":"2025-02-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143315862","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-04DOI: 10.1016/j.molimm.2025.01.013
Li Ding , Huijun Lin , Zhidong Ma , Yong He , Sheng Ding , Kaile Zhang , Jiechao Zhang , Wenyao Li , Lianbo Xiao
Stigmasterol (Stig), a phytosterol with anti-inflammatory and antioxidant properties, has been shown to have potential therapeutic effects. In this study, we aimed to investigate whether Stig mitigates rheumatoid arthritis (RA) progression by reducing chondrocyte injury. A mouse model of RA was established by intradermally injecting type II collagen into the tail roots of mice. The arthritic score and spleen index were measured in RA mice to assess the effects of Stig on RA progression. Lipopolysaccharide (LPS)-treated chondrocytes were used as a cellular model of RA. The roles of Stig in chondrocyte viability, proliferation, migration, inflammation, and injury were assessed using Cell Counting Kit-8, EdU, Transwell assays, quantitative real-time PCR, and western blotting, respectively. The results demonstrated that Stig exhibited no significant cytotoxicity in CHON-001 chondrocytes. Interestingly, it effectively inhibited LPS-induced apoptosis and increased cell viability, proliferation, and migration. Stig also alleviated LPS-induced pro-inflammatory responses and CHON-001 cell injury. Mechanistically, Stig activated nuclear factor erythroid 2-related factor 2 (Nrf2) signaling, which led to the inactivation of the NOD-like receptor protein 3 (NLRP3) inflammasome and a subsequent decrease in CHON-001 cell pyroptosis. However, the protective effects of Stig were abrogated by ML385, a specific inhibitor Nrf2. Stig treatment further improved the clinical severity in RA mice. In summary, Stig reduces LPS-induced chondrocyte injury and mitigates RA progression by inhibiting Nrf2/NLRP3-mediated pyroptosis, offering a potential therapeutic approach for RA.
{"title":"Stigmasterol mitigates rheumatoid arthritis progression by decreasing Nrf2/NLRP3-mediated pyroptosis in chondrocyte","authors":"Li Ding , Huijun Lin , Zhidong Ma , Yong He , Sheng Ding , Kaile Zhang , Jiechao Zhang , Wenyao Li , Lianbo Xiao","doi":"10.1016/j.molimm.2025.01.013","DOIUrl":"10.1016/j.molimm.2025.01.013","url":null,"abstract":"<div><div>Stigmasterol (Stig), a phytosterol with anti-inflammatory and antioxidant properties, has been shown to have potential therapeutic effects. In this study, we aimed to investigate whether Stig mitigates rheumatoid arthritis (RA) progression by reducing chondrocyte injury. A mouse model of RA was established by intradermally injecting type II collagen into the tail roots of mice. The arthritic score and spleen index were measured in RA mice to assess the effects of Stig on RA progression. Lipopolysaccharide (LPS)-treated chondrocytes were used as a cellular model of RA. The roles of Stig in chondrocyte viability, proliferation, migration, inflammation, and injury were assessed using Cell Counting Kit-8, EdU, Transwell assays, quantitative real-time PCR, and western blotting, respectively. The results demonstrated that Stig exhibited no significant cytotoxicity in CHON-001 chondrocytes. Interestingly, it effectively inhibited LPS-induced apoptosis and increased cell viability, proliferation, and migration. Stig also alleviated LPS-induced pro-inflammatory responses and CHON-001 cell injury. Mechanistically, Stig activated nuclear factor erythroid 2-related factor 2 (Nrf2) signaling, which led to the inactivation of the NOD-like receptor protein 3 (NLRP3) inflammasome and a subsequent decrease in CHON-001 cell pyroptosis. However, the protective effects of Stig were abrogated by ML385, a specific inhibitor Nrf2. Stig treatment further improved the clinical severity in RA mice. In summary, Stig reduces LPS-induced chondrocyte injury and mitigates RA progression by inhibiting Nrf2/NLRP3-mediated pyroptosis, offering a potential therapeutic approach for RA.</div></div>","PeriodicalId":18938,"journal":{"name":"Molecular immunology","volume":"179 ","pages":"Pages 9-17"},"PeriodicalIF":3.2,"publicationDate":"2025-02-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143255856","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-01DOI: 10.1016/j.molimm.2025.01.001
Zijie Li , Xiaokuan Zhang , Yuying Qi , Zhiyu Wang
Esophageal squamous cell carcinoma (ESCC) is a common malignancy. Programmed death ligand 1 of small extracellular vesicles (sEV-PDL1) induce immune evasion and enhance tumor progression. However, the role of ESCC derived sEV-PDL1 in modulating CD8+T cell remains unclear. sEVs were isolated through differential centrifugation. CD8+T cells were isolated, stimulated and cultured with sEVs to evaluate the proportions, phenotypes, and functions by flow cytometry. Lentivirus infection and Crisper-Cas9 were used to constructed stable transgenic cell lines: Eca109-PDL1kd and mEC25-PDL1ko. The proportions of CD8+T cells in ESCC patients was lower than healthy donors (HD). Furthermore, a negative correlation between sEV-PDL1 and CD8+T cell was observed. sEV-PDL1 induced CD8+T cell exhaustion by reducing the expression levels of Ki67, Granzyme B (GrzmB), and interferon-γ (IFN-γ) both in vitro and in vivo. However, anti-PDL1 reversed the result. Our findings reveal that targeting sEV-PDL1 to rejuvenate CD8+T cell functions is one of the mechnisms a promising therapeutic strategy for ESCC.
{"title":"Esophageal squamous cell carcinoma derived sEV-PDL1 exhausts CD8+T cells to promote immunosuppression","authors":"Zijie Li , Xiaokuan Zhang , Yuying Qi , Zhiyu Wang","doi":"10.1016/j.molimm.2025.01.001","DOIUrl":"10.1016/j.molimm.2025.01.001","url":null,"abstract":"<div><div>Esophageal squamous cell carcinoma (ESCC) is a common malignancy. Programmed death ligand 1 of small extracellular vesicles (sEV-PDL1) induce immune evasion and enhance tumor progression. However, the role of ESCC derived sEV-PDL1 in modulating CD8<sup>+</sup>T cell remains unclear. sEVs were isolated through differential centrifugation. CD8<sup>+</sup>T cells were isolated, stimulated and cultured with sEVs to evaluate the proportions, phenotypes, and functions by flow cytometry. Lentivirus infection and Crisper-Cas9 were used to constructed stable transgenic cell lines: Eca109-PDL1<sup>kd</sup> and mEC25-PDL1<sup>ko</sup>. The proportions of CD8<sup>+</sup>T cells in ESCC patients was lower than healthy donors (HD). Furthermore, a negative correlation between sEV-PDL1 and CD8<sup>+</sup>T cell was observed. sEV-PDL1 induced CD8<sup>+</sup>T cell exhaustion by reducing the expression levels of Ki67, Granzyme B (GrzmB), and interferon-γ (IFN-γ) both in vitro and in vivo. However, anti-PDL1 reversed the result. Our findings reveal that targeting sEV-PDL1 to rejuvenate CD8<sup>+</sup>T cell functions is one of the mechnisms a promising therapeutic strategy for ESCC.</div></div>","PeriodicalId":18938,"journal":{"name":"Molecular immunology","volume":"178 ","pages":"Pages 12-19"},"PeriodicalIF":3.2,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142971520","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-01DOI: 10.1016/j.molimm.2025.01.004
Shuncai Bao , Guangpeng Li , Xue Lu , Tengfei Lu , Xiaohui Hou
Background
Midges are widely distributed globally. They can transmit numerous serious diseases as well as trigger an allergic reaction in the host. Their saliva contains a variety of proteins that act as sensitizers to stimulate the host's immune response, leading to IgE-mediated allergic symptoms.
Material and method
In the present study, we constructed a mouse sensitization model with the thorax extract of the Culicoides tainanus, and evaluated the sensitization model by behavior and specific antibody expression. SDS-PAGE/western blot was used to detect the binding proteins by IgE antibody in the thorax extract of the C. tainanus. The objective band was cut for mass spectrometry to preliminarily clarify the potential allergen. The pET21a-Cul t 1 recombinant expression vector was constructed and the target protein was purified by Ni affinity chromatography. The sensitizing effect of the sensitizer was verified in vitro and in vivo.
Results
Immunoblot analysis revealed that a 66 kDa protein (Cul t 1) from the chest extracts of C. tainanus could bind to serum IgE of sensitized mice. Cul t 1 was further identified by fragmentation, mass spectrometry and bioinformatics as maltase, an enzyme involved in sugar digestion. In vivo validation of the Cul t 1-mouse sensitization model showed that the scratching behavior of the Cul t 1 sensitized group was significantly higher than that of the control group according to behavioral evaluation. The results of HE staining of the skin of the injected area showed that Cul t 1 sensitized group was found to have a large number of inflammatory cell infiltration and increased fibrosis in the dermis. The ELISA detection of IgE showed that the Cul t 1 sensitized group was significantly elevated from the beginning of the 28th day onwards, and the expression level of IgE had a tendency to slow down with the prolongation of the sensitization time. The ELISA assay showed that the expression level of IgG1 increased significantly in the Cul t 1 group on day 42, while the results of the specific antibody IgG2a showed that there was no significant difference in both the Cul t 1 sensitized group and the control group. The results suggest that the immune response induced by Cul t 1 sensitized mice may be shifted toward a Th2 type immune response.
Conclusions
In this study, we identified the sensitizer of the C. tainanus, named Cul t 1. We successfully constructed the recombinant expression vector pET21a-Cul t 1, purified the Cul t 1 sensitizer by affinity chromatography, and verified the sensitizing effect of Cul t 1 in mice. The results laid a practicable foundation for the subsequent immune therapy of midge bites and bite control.
{"title":"Identification and characterization of the Cul t 1 as major allergen from biting midge Culicoides tainanus","authors":"Shuncai Bao , Guangpeng Li , Xue Lu , Tengfei Lu , Xiaohui Hou","doi":"10.1016/j.molimm.2025.01.004","DOIUrl":"10.1016/j.molimm.2025.01.004","url":null,"abstract":"<div><h3>Background</h3><div>Midges are widely distributed globally. They can transmit numerous serious diseases as well as trigger an allergic reaction in the host. Their saliva contains a variety of proteins that act as sensitizers to stimulate the host's immune response, leading to IgE-mediated allergic symptoms.</div></div><div><h3>Material and method</h3><div>In the present study, we constructed a mouse sensitization model with the thorax extract of the <em>Culicoides tainanus</em>, and evaluated the sensitization model by behavior and specific antibody expression. SDS-PAGE/western blot was used to detect the binding proteins by IgE antibody in the thorax extract of the <em>C. tainanus</em>. The objective band was cut for mass spectrometry to preliminarily clarify the potential allergen. The pET21a-<em>Cul t 1</em> recombinant expression vector was constructed and the target protein was purified by Ni affinity chromatography. The sensitizing effect of the sensitizer was verified <em>in vitro</em> and <em>in vivo</em>.</div></div><div><h3>Results</h3><div>Immunoblot analysis revealed that a 66 kDa protein (Cul t 1) from the chest extracts of <em>C. tainanus</em> could bind to serum IgE of sensitized mice. Cul t 1 was further identified by fragmentation, mass spectrometry and bioinformatics as maltase, an enzyme involved in sugar digestion. <em>In vivo</em> validation of the Cul t 1-mouse sensitization model showed that the scratching behavior of the Cul t 1 sensitized group was significantly higher than that of the control group according to behavioral evaluation. The results of HE staining of the skin of the injected area showed that Cul t 1 sensitized group was found to have a large number of inflammatory cell infiltration and increased fibrosis in the dermis. The ELISA detection of IgE showed that the Cul t 1 sensitized group was significantly elevated from the beginning of the 28th day onwards, and the expression level of IgE had a tendency to slow down with the prolongation of the sensitization time. The ELISA assay showed that the expression level of IgG1 increased significantly in the Cul t 1 group on day 42, while the results of the specific antibody IgG2a showed that there was no significant difference in both the Cul t 1 sensitized group and the control group. The results suggest that the immune response induced by Cul t 1 sensitized mice may be shifted toward a Th2 type immune response.</div></div><div><h3>Conclusions</h3><div>In this study, we identified the sensitizer of the <em>C. tainanus</em>, named Cul t 1. We successfully constructed the recombinant expression vector pET21a-<em>Cul t 1</em>, purified the Cul t 1 sensitizer by affinity chromatography, and verified the sensitizing effect of Cul t 1 in mice. The results laid a practicable foundation for the subsequent immune therapy of midge bites and bite control.</div></div>","PeriodicalId":18938,"journal":{"name":"Molecular immunology","volume":"178 ","pages":"Pages 32-40"},"PeriodicalIF":3.2,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143008718","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-01DOI: 10.1016/j.molimm.2025.01.012
Yajie Hu , Xiaoli Deng , Yaming Lv , Chen Liu , Juan Chen , Jie Song , Yunhui Zhang
Neuronal death and neuroinflammation has been considered as the main contributors to the progression and deterioration of HFMD caused by CV-A10. Necroptosis is a lytic and inflammatory form of cell death that plays a crucial role in viral pathogenicity. Herein, our study showed that CV-A10-infected SH-SY5Y cells induced necroptosis via activating RIPK3-depedent pathway, but not requiring RIPK1, and meanwhile triggered the release of inflammatory cytokines. Moreover, RIPK3-mediated necroptosis was also involved in virus production, which did not require RIPK1 either. Finally, it was further verified that TLR3 drove RIPK3-mediated cell death by sensing CV-A10 RNA and activating RIPK3. Collectively, our study demonstrated that initiation of necroptosis in SH-SY5Y cells induced by CV-A10 accelerated the formation of inflammatory response and promoted virus replication through triggering a TLR3-initiated RIPK3-dependent pathway of necroptosis, which advanced the current understanding of necroptosis for the neuropathogenesis of CV-A10 infection.
{"title":"Coxsackievirus-A10 induced RIPK3-driven necroptosis to promote the formation of inflammatory response and enhance virus production via being recognized by TLR3","authors":"Yajie Hu , Xiaoli Deng , Yaming Lv , Chen Liu , Juan Chen , Jie Song , Yunhui Zhang","doi":"10.1016/j.molimm.2025.01.012","DOIUrl":"10.1016/j.molimm.2025.01.012","url":null,"abstract":"<div><div>Neuronal death and neuroinflammation has been considered as the main contributors to the progression and deterioration of HFMD caused by CV-A10. Necroptosis is a lytic and inflammatory form of cell death that plays a crucial role in viral pathogenicity. Herein, our study showed that CV-A10-infected SH-SY5Y cells induced necroptosis via activating RIPK3-depedent pathway, but not requiring RIPK1, and meanwhile triggered the release of inflammatory cytokines. Moreover, RIPK3-mediated necroptosis was also involved in virus production, which did not require RIPK1 either. Finally, it was further verified that TLR3 drove RIPK3-mediated cell death by sensing CV-A10 RNA and activating RIPK3. Collectively, our study demonstrated that initiation of necroptosis in SH-SY5Y cells induced by CV-A10 accelerated the formation of inflammatory response and promoted virus replication through triggering a TLR3-initiated RIPK3-dependent pathway of necroptosis, which advanced the current understanding of necroptosis for the neuropathogenesis of CV-A10 infection.</div></div>","PeriodicalId":18938,"journal":{"name":"Molecular immunology","volume":"178 ","pages":"Pages 107-116"},"PeriodicalIF":3.2,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143075077","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-01DOI: 10.1016/j.molimm.2025.01.006
Xinrong Li , Xiuhua Yao , Wei Zhao , Bo Wei , Ran Zhang , Geng Yan , Mingyu Ma , Zhenhai Wang , Xijun Liu , Yumei Liu , Guangyou Wang , Hulun Li , Qingfei Kong , Jinghua Wang , Lili Mu
As one of the largest organs of our human body, skeletal muscle has good research prospects in myasthenia gravis (MG), the symptoms of which include systemic skeletal muscle weakness. Skeletal muscle is composed of two types of muscle fibers. Different fiber subtypes can be converted into each other; however, the underlying mechanism is not yet clear. In this paper, we firstly established an experimental autoimmune myasthenia gravis (EAMG) rat model and found that the skeletal muscle fibers of the EAMG group were atrophied, with a change in the proportion of fiber subtypes, which switched from type IIa to type I in the EAMG group at the peak stage, as verified by histological and molecular analyses. Second-generation sequencing results predicted that the PI3K-Akt signaling pathway might be involved in the switch, and the mRNA expression levels of the PI3K-Akt pathway-related genesNr4a1, IL2rb, Col1A1 and Ddit4 were significantly different. In conclusion, this study indicates that the switch of muscle fiber subtypes in MG via the PI3K-Akt signaling pathway may be a potential target for the treatment of MG-related skeletal muscle atrophy in the future.
{"title":"Muscle fiber types switched during the development of experimental autoimmune myasthenia gravis via the PI3K/Akt signaling pathway","authors":"Xinrong Li , Xiuhua Yao , Wei Zhao , Bo Wei , Ran Zhang , Geng Yan , Mingyu Ma , Zhenhai Wang , Xijun Liu , Yumei Liu , Guangyou Wang , Hulun Li , Qingfei Kong , Jinghua Wang , Lili Mu","doi":"10.1016/j.molimm.2025.01.006","DOIUrl":"10.1016/j.molimm.2025.01.006","url":null,"abstract":"<div><div>As one of the largest organs of our human body, skeletal muscle has good research prospects in myasthenia gravis (MG), the symptoms of which include systemic skeletal muscle weakness. Skeletal muscle is composed of two types of muscle fibers. Different fiber subtypes can be converted into each other; however, the underlying mechanism is not yet clear. In this paper, we firstly established an experimental autoimmune myasthenia gravis (EAMG) rat model and found that the skeletal muscle fibers of the EAMG group were atrophied, with a change in the proportion of fiber subtypes, which switched from type IIa to type I in the EAMG group at the peak stage, as verified by histological and molecular analyses. Second-generation sequencing results predicted that the PI3K-Akt signaling pathway might be involved in the switch, and the mRNA expression levels of the PI3K-Akt pathway-related genesNr4a1, IL2rb, Col1A1 and Ddit4 were significantly different. In conclusion, this study indicates that the switch of muscle fiber subtypes in MG via the PI3K-Akt signaling pathway may be a potential target for the treatment of MG-related skeletal muscle atrophy in the future.</div></div>","PeriodicalId":18938,"journal":{"name":"Molecular immunology","volume":"178 ","pages":"Pages 41-51"},"PeriodicalIF":3.2,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143008720","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}