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Spatiotemporal regulation of ubiquitin-mediated protein degradation via upconversion optogenetic nanosystem

Nano Research 2020年第12期13卷 3253-3260页

作者： Yafeng Hao Taofeng Du gaoju pang Jiahua Li Huizhuo Pan Yingying Zhang Lizhen Wang Jin Chang En-min Zhou Hanjie Wang School of Life Sciences Tianjin UniversityTianjin300072China Department of preventive Veterinary Medicine College of Veterinary MedicineNorthwest A&F UniversityYangling712100China

Protein degradation technology,which is one of the most direct and effective ways to regulate the life activities of cells,is expected to be applied to the treatment of various ***,current protein degradation technologies such as some small-molecule degraders which are unable to achieve spatiotemporal regulation,making them difficult to transform into clinical *** this article,an upconversion optogenetic nanosystem was designed to attain accurate regulation of protein *** system worked via two interconnected parts:1)the host cell expressed light-sensitive protein that could trigger the ubiquitinproteasome pathway upon blue-light exposure;2)the light regulated light-sensitive protein by changing light conditions to achieve regulation of protein *** results based on model protein(Green Fluorescent Protein,GFP)validated that this system could fulfill protein degradation both in vitro(both Hela and 293T cells)and in vivo(by upconversion optogenetic nanosystem),and further demonstrated that we could reach spatiotemporal regulation by changing the illumination time(0–25 h)and the illumination frequency(the illuminating frequency of 0–30 s every 1 min).We further took another functional protein(The Nonstructural Protein 9,NSP9)into *** confirmed that the proliferation of porcine reproductive and respiratory syndrome virus(PRRSV)was inhibited by degrading the NSP9 in this light-induced system,and PRRSV proliferation was affected by different light conditions(illumination time varies from 0–24 h).We expected this system could provide new perspectives into spatiotemporal regulation of protein degradation and help realize the clinical application transformation for treating diseases of protein degradation technology.

关键词： protein degradation ubiquitin-proteasome system optogenetics upconversion materials the nonstructural protein9(NSP9) porcine reproductive and respiratory syndrome virus(PRRSV)

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A micro-nano optogenetic system based on probiotics for in situ host metabolism regulation

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Nano Research 2023年第2期16卷 2829-2839页

作者： Xinyu Zhang Ning Ma Wei Ling gaoju pang Tao Sun Jing Liu Huizhuo Pan Meihui Cui Chunli Han Chun Yang Jin Chang Xian Huang Hanjie Wang School of Life Sciences Tianjin UniversityTianjin 300072China School of Precision Instrument and Optoelectronics Engineering Tianjin UniversityTianjin 300072China School of Chemical Engineering&Technology Tianjin UniversityTianjin 300072China

Genetically engineered bacteria have aroused attention as micro-nano drug delivery systems in ***,conventional designs of engineered bacteria usually function constantly or autonomously,which might be non-specific or ***,designing and optimizing in situ control strategy are important methodological progress for therapeutic researches of intestinal engineered ***,a micro-nano optogenetic system based on probiotic was developed combining microelectronics,nanotechnology,and synthetic biology to achieve in situ controllable drug ***,optogenetic engineered Lactococcus lactis was orally administrated in the intestinal tract.A wearable optical device was designed to control optical signals ***,*** could be customized to secrete peptides according to optical *** an example,optogenetic *** system can be constructed to secrete glucagon-like peptide-1(GLP-1)under the control of the wearable optical device to regulate *** improve the half-life of GLP-1 in vivo,Fc-domain fused GLP-1 was optimally *** this strategy,blood glucose,weight,and other features were well controlled in rats and mice ***,upconversion microcapsules were introduced to increase the excitation wavelength of the optogenetic system for better *** strategy has biomedical potential to expand the toolbox for intestinal engineered bacteria.

关键词： synthetic biology optogenetics upconversion nanoparticles microelectronics diabetics

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IL-21/IL-21R regulates the neutrophil-mediated pathologic immune response during chlamydial respiratory infection

IL-21/IL-21R regulates the neutrophil-mediated pathologic im...

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第十四届全国免疫学学术大会

作者： Jiajia Zeng Yueyue Xu Lu Tan Xiaoyu Zha Shuaini Yang Hong Zhang Yuqing Tuo Wenhao Niu gaoju pang Lida Sun Hong Bai Tianjin Medical University

IL-21/IL-21R was documented to have pleiotropic regulatory effects on several infection and inflammation. However, the role and mechanism of IL-21/IL-21R on immune response during chlamydial respiratory infection remain unknown. In this study, the regulation of IL-21/IL-21R on neutrophilic inflammation was investigated in mice model of Chlamydia muridarum(C. muridarum) respiratory infection. Our Results showed increased IL-21 and IL-21R mRNA expression in C. muridarum-infected lungs of C57BL/6 wild-type(WT) mice and protective immunity was observed in IL-21Rmice with less body weight loss, increased clearance of pulmonary C. muridarum and milder pulmonary pathology. Further analysis showed lower infiltration, CD64 expression, MPO activity and neutrophil-produced proteins production of pulmonary neutrophils in IL-21Rmice compared with WT mice after infection. These Results correlated well with the decrease of neutrophil related chemokines(KC and MIP-2), inflammatory cytokines(IL-6, IL-1β and TNF-α) and TLR/MyD88 pathway mediators(TLR2, TLR4 and MyD88) in lungs of IL-21Rmice. Complementarily, we used rIL-21 treatment to raise IL-21/IL-21R effect and observed enhanced promoting effects on disease development, neutrophilic inflammation, chemotactic factors and TLR/MyD88 signal in C. muridarum-infected mice. These Results demonstrated that IL-21/IL-21R aggravates the neutrophil inflammation through regulating TLR/MyD88 signal pathway during chlamydial respiratory infection.

关键词： chlamydia IL-21 neutrophil pathological mechanism

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IL-27/IL-27R promotes host defense against chlamydial lung infection by suppressing neutrophils recruitment mediated by Th17

IL-27/IL-27R promotes host defense against chlamydial lung i...

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第十三届全国免疫学学术大会

作者： Xiaoyu zha gaoju pang Lida Sun Sai Qiao Wenhao Niu Juyou Liang Lu Tan Hong Zhang Tengli Liu Huili Zhao Yue Wang Hong Bai Tianjin Medical University

Interleukin-27, a heterodimeric cytokine of IL-12 family, composed of Epstein-Barr virus-induced gene 3(EBI3) and p28, has diverse influences on the immune response. In this study, we identified the role of IL-27/IL-27 R in host defense against Chlamydia lung infection and further investigated the influence of IL-27/IL-27 R on Th1 and Th17 cell responses as well as neutrophils recruitment. Our data showed that mRNA expression of IL-27(EBI3) and IL-27 R(WSX-1) were significantly induced after Cm infection especially on day 7 post-infection(p.i.) in wild type mice. Mice with WSX-1 knockout(WSX-1mice) showed higher severity of the disease with greater body weight loss, more chlamydia growth and more severe inflammatory cell infiltration in lungs compared with WT mice. Th1 cell responses, which played a protective role in Cm infection, was significantly weaken in lungs on day 7 p.i. and day 14 p.i. whereas Th17 cell responses, which can promote neutrophil recruitment, was dramatically enhanced in lungs on day 7 p.i. and day 14 p.i. in WSX-1mice compared with WT mice. Neutrophils in lung were also increased on day 7 p.i. and day 14 p.i. in WSX-1mice compared to WT mice which was in accordant with upregulated mRNA expression of chemokines and cytokines of neutrophils including IL-17, TNFα, IL-6 and CXCR2. To further confirm that severe disease condition in WSX-1mice was caused by increased IL-17 in lungs, we found that WSX-1mice injected with mouse IL-17/IL-17 A-blocking antibody on the day before infection, day 2 p.i. and day 4 p.i. showed improved disease status in the aspects of body weight change, chlamydia growth and neutrophil recruitment in lungs compared to those without IL-17 blocking. Taken together, we concluded that IL-27/IL-27 R plays a protective role in host defense against Chlamydia lung infection by suppressing neutrophils recruitment mediated by Th17.

关键词： Chlamydia Lung infection IL-27/IL-27R Th17 Neutrophil

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Vγ1+T and Vγ4+T cells mediate majority of γδT cell function during chlamydial airway infection

Vγ1+T and Vγ4+T cells mediate majority of γδT cell funct...

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第十二届全国免疫学学术大会

作者： sun lida pang gaoju Wang Yue Tang Yingying Qiao Sai Liang Juyou Zhao Huili Liu Tengli Zheng Ningbo Tan Lu Zhang Yongci Zhang Hong Bai Hong Department of Immunology School of Basic Medical SciencesTianjin Medical UniversityTianjin Key Laboratory of Cellular and Molecular ImmunologyKey Laboratory of Educational Ministry of China

Our previous studies have shown that γδT cells plays an immune protective role against Chlamydial muridarum(Cm)lung infection by producing IFNγ and IL-17 at the early stage of infection and mice deficient in TCRγδ developed more severe disease compared to the wild type(WT) ***,the γδT cells subsets and their functions in immune protection against Cm infection remain ***,We investigate the proliferation,activation and function of lung y8 T cells subsets following Cm lung *** mice were inoculated intranasally with 1×10 inclusion-forming units(IFUs) of Cm to induce the murine Chlamydia *** y8 T cells subsets in lung mononuclear cells were analysed by using flow cytometry and *** cytokine staining was used to detect IFNγ and IL-17 secretion levels by Vγ1T,Vγ4T *** results showed that there are 5 kinds of γδT cell subsets including Vγ1T,Vγ2T,Vγ4T,Vγ5T and Vγ6T cells in lungs of naive *** percentage of each subpopulation shows as Vγ2 > Vγ4 > Vγ1> Vγ6 > Vγ5 in WT ***,Cm lung infection mainly induced Vγ4T cells proliferation in early stage,and then Vγ1T cells became the main γδT cell subsets gradually.Vγ6T also increased after infection and proliferate significantly on 7 th day,however Vγ2T and Vγ5T cells had no significant change in the lung after Cm *** detection showed Vγ1T cells could secrete IFNγ but do not product IL-17,and Cm infection induced IFNγ secretion reached the peak on day 7(p<0.01).Vγ4T cells could secrete both IFNγ and IL-17,and the cytokines levels increased rapidly after Cm infection and reached the peak on day 3(p<0.001).These findings provide in vivo evidence that Vγ4T cells is the major IL-17 and IFNγ-producing γδT cell subsets at the early period of Cm airway infection.

关键词： Chlamydia muridarum γδT cells subsets IL-17 IFNγ

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The subset characteristics of γδT cells in the lung of mice following Chlamydial muridarum airway infection

The subset characteristics of γδT cells in the lung of mic...

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第十一届全国免疫学学术大会

作者： sun lida pang gaoju Wang Yue Tang Yingying Qiao Sai Liang Juyou Zhao Huili iu Tengli Zheng Ningbo Tan Lu Zhang Yongci Zhang Hong Bai Hong Department of Immunology School of Basic Medical SciencesTianjin Medical UniversityTianjin Key Laboratory of Cellular and Molecular ImmunologyKey Laboratory of Educational Ministry of China

Our previous studies have shown that γδT cells plays an immune protective role against Chlamydial muridarum(Cm) lung infection by producing INFγ and IL-17 at the early stage of infection. However, the γδT cells subsets and their functions in immune protection against Cm infection remain elusive. Here, We investigate the proliferation, activation and function of lung γδT cells subsets following Cm lung *** mice were inoculated intranasally with 1×10 inclusion-forming units(IFUs) of Cm to induce the murine Chlamydia pneumonia. The γδT cells subsets in lung mononuclear cells were analysed by using flow cytometry and RT-PCR, intra-cellular cytokine staining was used to detect IFNγ and IL-17 secretion levels of Vγ1T, Vγ4T cells. Our results showed that there are 5 kinds of γδT cell subsets including Vγ1T、Vγ2T、Vγ4T、Vγ5T and Vγ6T cells in lung of mice and the contents are compared to Vγ2 > Vγ4 > Vγ1 > Vγ6 > Vγ5 in WT mice. Remarkably, Cm lung infection mainly induced Vγ4T cells proliferation in early stage, and then Vγ1T cells became the main γδT cell subsets gradually. Vγ6T also increased after infection and proliferate significantly on 7th day, however Vγ2T and Vγ5T cells had no significant change in the lung after Cm infection. Cytokines detection showed Vγ1T cells could secrete IFNγ but do not product IL-17, and Cm infection induced IFNγ secretion reached the peak on 7th day(p<0.01). Vγ4T cells could secrete IFNγ and IL-17, and the cytokines levels increased rapidly after Cm infection and reached the peak on 3th day(p<0.001). These findings provide in vivo evidence that Vγ4T cells is the major IL-17 and IFNγ-producing γδT cell subsets at the early period of Cm airway infection.

关键词： Chlamydia muridarum γδT cells subsets IL-17 IFNγ

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沙眼衣原体呼吸道感染诱导PI3K/AKT/Bcl-2/Bax通路活化促进CD4^＋ T细胞凋亡

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中华微生物学和免疫学杂志 2017年第3期37卷 178-182页

作者：檀露郑佞波梁聚友孙丽妲庞高举张虹白虹天津医科大学基础医学院免疫学系.国家教育部免疫微环境与疾病重点实验室天津市细胞和分子免疫学重点实验室300070

目的探讨PI3K/AKT/Bcl-2/Bax途径在小鼠沙眼衣原体呼吸道感染中的诱导活化及CD4^＋T细胞的凋亡。方法C57BL/6小鼠，经鼻腔吸入1×10^3 IFU沙眼衣原体小鼠肺炎菌株（Cm），诱导小鼠衣原体肺炎。于感染的不同时间点分离小鼠肺组织单个核... 详细信息

目的探讨PI3K/AKT/Bcl-2/Bax途径在小鼠沙眼衣原体呼吸道感染中的诱导活化及CD4^＋T细胞的凋亡。方法C57BL/6小鼠，经鼻腔吸入1×10^3 IFU沙眼衣原体小鼠肺炎菌株（Cm），诱导小鼠衣原体肺炎。于感染的不同时间点分离小鼠肺组织单个核细胞，RT-PCR检测PI3K p110δ mRNA表达；Western blot检测p-AKT、Bcl-2、Bax蛋白表达；流式细胞术检测不同感染时间小鼠肺组织CD4^＋ T细胞的凋亡。结果与未感染组比较，小鼠Cm呼吸道感染诱导感染肺组织PI3K p110δ mRNA表达及PI3K下游分子p-AKT蛋白表达增高，两者均于感染第3天显著增高，第7天达到高峰，感染第14天有所降低。同时，Cm呼吸道感染也诱导凋亡相关蛋白Bcl-2与Bax蛋白表达显著增高，均于第7天达到高峰，但Bcl-2/Bax比值依感染时间降低，提示抗凋亡作用逐渐降低，细胞凋亡增加。Cm感染诱导小鼠肺组织CD4^＋T细胞的凋亡，于感染第3天早期凋亡（Annexin＋PI＋）和细胞总凋亡（Annexin＋PI）均开始增高（P〈0.05），感染第7天达高峰（P〈0.001）。结论Cm小鼠呼吸道感染诱导PI3K/AKT/Bcl-2/Bax通路活化导致CD4^＋ T细胞凋亡。

关键词：沙眼衣原体 P13K/AKT CD4^+T细胞凋亡

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衣原体呼吸道感染早期IL-17对调节性T细胞及其IL-10分泌的影响

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中华微生物学和免疫学杂志 2018年第11期38卷 807-811页

作者：张虹庞高举查晓宇钮文豪刘腾丽赵慧丽白虹天津医科大学基础医学院免疫学系国家教育部免疫微环境与疾病重点实验室天津市细胞和分子免疫学重点实验室300070

目的探讨IL-17在沙眼衣原体呼吸道感染中对调节性T细胞的影响.方法鼻滴给予野生型(WT,C57BL/6)小鼠和 IL-17-/-小鼠 1×10^3包涵体形成单位(IFU)的鼠型衣原体(Chlamydia muridarum, Cm),构建小鼠衣原体呼吸道感染模型.制备小鼠脾、肺... 详细信息

目的探讨IL-17在沙眼衣原体呼吸道感染中对调节性T细胞的影响.方法鼻滴给予野生型(WT,C57BL/6)小鼠和 IL-17-/-小鼠 1×10^3包涵体形成单位(IFU)的鼠型衣原体(Chlamydia muridarum, Cm),构建小鼠衣原体呼吸道感染模型.制备小鼠脾、肺组织单个核细胞悬液,运用流式细胞术检测调节性T细胞(Treg)百分率;利用RT-PCR技术检测小鼠肺组织中Foxp3和TGF-β的mRNA表达水平;用ELISA方法检测小鼠支气管肺泡灌洗液中IL-10的产生.结果衣原体呼吸道感染第3天,IL-17--小鼠脾、肺组织中的CD4+CD25+T细胞与CD4+CD25+Foxp3+T细胞比例均高于WT小鼠;IL-17-/-小鼠肺组织Foxp3 mRNA表达及支气管肺泡灌洗液中IL-10的分泌显著高于WT小鼠;两种小鼠肺组织TGF-β mRNA表达无显著差异.结论衣原体呼吸道感染早期阶段IL-17可能抑制小鼠调节性T细胞的增殖和IL-10的产生.

关键词：衣原体 Treg IL-17 Th17

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产生IL-17的γδT细胞在沙眼衣原体呼吸道感染早期促进中性粒细胞的募集

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中华微生物学和免疫学杂志 2017年第1期37卷 1-5页

作者：梁聚友孙丽妲庞高举王悦乔赛檀露张虹唐莹莹刘腾丽赵慧丽白虹天津医科大学基础医学院免疫学系、国家教育部免疫微环境与疾病重点实验室、天津市细胞和分子免疫学重点实验室 300070

目的探讨沙眼衣原体呼吸道感染早期产生IL-17的γδT细胞对中性粒细胞募集的影响及其机制。方法WT（C57BL/6）与TCRδ－/－小鼠鼻腔吸入3×103 IFU（inclusion-forming units）沙眼衣原体小鼠肺炎菌株（Chlamydia muridarum, Cm），建... 详细信息

目的探讨沙眼衣原体呼吸道感染早期产生IL-17的γδT细胞对中性粒细胞募集的影响及其机制。方法WT（C57BL/6）与TCRδ－/－小鼠鼻腔吸入3×103 IFU（inclusion-forming units）沙眼衣原体小鼠肺炎菌株（Chlamydia muridarum, Cm），建立沙眼衣原体小鼠肺炎模型。分离感染不同时间点小鼠肺组织单个核细胞，流式细胞术检测肺组织CD11b＋Gr1＋中性粒细胞（PMN）百分率；细胞内因子染色检测γδT细胞IL-17的合成；RT-PCR检测小鼠肺组织IL-17及KC（中性粒细胞趋化因子）、IL-6 mRNA表达。结果一定剂量的Cm呼吸道感染诱导小鼠肺组织γδT细胞分泌大量IL-17，与未感染组比较，感染后第3天小鼠肺组织CD3＋ γδT＋细胞分泌的IL-17显著增加（P〈0.001），并且持续到感染的第7天（P〈0.05）；TCRδ－/－小鼠Cm呼吸道感染后第3天肺组织IL-17 mRNA的表达以及PMN百分率显著低于WT鼠；进一步研究显示TCRδ－/－小鼠在感染后第3天及第7天中性粒细胞趋化因子KC和IL-6在小鼠肺组织的表达显著降低。结论产生大量IL-17的γδT细胞可能通过上调KC和IL-6的表达在沙眼衣原体呼吸道感染中发挥PMN募集作用。

关键词：沙眼衣原体肺炎菌株中性粒细胞 γδT细胞 IL-17

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