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Repurposing Loperamide as an Anti-Infection Drug for the Treatment of Intracellular Bacterial Pathogens

Engineering 2024年第8期39卷 180-193页

作者： Hongtao Liu Siqi Li Le Deng Zhenxu Shi Chenxiao Jiang Jingyan Shu Yuan Liu Xuming Deng Jianfeng Wang Zhimin Guo Jiazhang Qiu State key laboratory for Diagnosis and Treatment of Severe Zoonotic infectious diseases&key laboratory of zoonosis Research Ministry of EducationInstitute of ZoonosisCollege of Veterinary Medicine&Jilin UniversityChangchun 130062China center for Pathogen Biology and infectious diseases The First Hospital of Jilin UniversityChangchun 130031China jiangsu co-innovation center for prevention and control of important animal infectious diseases and zoonoses Institute of Comparative MedicineCollege of Veterinary MedicineYangzhou UniversityYangzhou 225009China

Infections caused by intracellular bacterial pathogens are difficult to treat since most antibiotics have low cell permeability and undergo rapid degradation within *** rapid development and dissemination of antimicrobial–resistant strains have exacerbated this *** the increasing knowledge of host–pathogen interactions,especially bacterial strategies for survival and proliferation within host cells,host-directed therapy(HDT)has attracted increased interest and has emerged as a promising antiinfection method for treating intracellular ***,we applied a cell-based screening approach to a US Food and Drug Administration(FDA)-approved drug library to identify compounds that can inhibit the intracellular replication of Salmonella Typhimurium(***).This screening allowed us to identify the antidiarrheal agent loperamide(LPD)as a potent inhibitor of *** intracellular *** treatment of infected cells markedly promoted the host autophagic response and lysosomal activity.A mechanistic study revealed that the increase in host autophagy and elimination of intracellular bacteria were dependent on the high expression of glycoprotein nonmetastatic melanoma protein B(GPNMB)induced by *** addition,LPD treatment effectively protected against *** infection in Galleria mellonella and mouse ***,our study suggested that LPD may be useful for the treatment of diseases caused by intracellular bacterial ***,LPD may serve as a promising lead compound for the development of anti-infection drugs based on the HDT strategy.

关键词： Intracellular bacteria US Food and Drug Administration(FDA)-approved drugs Drug repurposing Loperamide Autophagy Glycoprotein nonmetastatic melanoma protein B

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Mesoporous SiO_(2) Sphere-Based Electrochemical Impedance Immunosensor for Ultrasensitive Detection of Bovine Interferon-γ

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Journal of Analysis and Testing 2023年第3期7卷 295-303页

作者： Yanping Xia Hui Chen Ruixin Liu Feng Shi Chuanli Ren Juan Li Jie Zhao Xiang Chen Zhanjun Yang School of Chemistry and Chemical Engineering Yangzhou UniversityYangzhou 225002China jiangsu key lab of zoonosis/jiangsu co-innovation center for prevention and control of important animal infectious diseases and zoonoses Yangzhou UniversityYangzhou 225009China Department of Geriatrics Clinical Medical College of Yangzhou UniversityYangzhou 225001China

Interferon-γis a kind of protein with a wide range of biological activities,which can regulate the immune function of the body,and can be used as an important marker to detect and treat bovine tuberculosis ***,a picogram-level bovine interferon-γ(BoIFN-γ)electrochemical impedance immunosensor was constructed for the first time using mesoporous silica nanospheres(MS Ns)to immobilize specific monoclonal BoIFN-y *** MS Ns and BoIFN-γimmuno sensors were characterized using scanning electron microscopy,transmission electron microscope,nitrogen adsorption experiment,X-ray photoelectron spectra,and contact angle *** possess a substantial specific surface area and significant hydrophilicity,and can immobilize many antibody molecules,thereby improving detection *** immunosensor has a linear detection range from 0.001 to 10.0 ng/mL with an exceptionally low detection limit of 0.62 pg/*** to the traditional BoIFN-γanalysis method,BoIFN-γimmunosensor presents superiorities in sensitivity,wide linear range as well as short processing *** importantly,the BoIFN-γsensor exhibits high selectivity,reliable repeatability as well as stability,providing a promising application prospect for the early diagnosis of Mycobacterium bovis infection.

关键词： Interferon-γ Electrochemical immunosensor Mesoporous SiO_(2) label-free Mycobacterium bovis

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Amino acids 89-96 of Salmonella typhimunum Tlagell=n represent the major domain responsible for TLR5-independent adjuvanticity in the humoral immune response

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Cellular & Molecular Immunology 2015年第5期12卷 625-632页

作者： Lei Zhang Zhiming Pan Xilong Kang Yun Yang Heekap Kang Na Zhang James M Rosati Xinan Jiao jiangsu co-innovation center for prevention and control of important animal infectious diseases and zoonosis Yangzhou China jiangsu key laboratory of zoonosis Yangzhou University Yangzhou China Ministry of Education key lab for Avian Preventive Medicine Yangzhou University Yangzhou China Division of Nephrology and Hypertension Department of Medicine Northwestern University Feinberg School of Medicine Chicago IL USA

Toll-like receptor 5 （TLR5） signaling in response to flagellin is dispensable for inducing humoral immunity, but alterations of aa 89-96, the TLR5 binding site, significantly reduced the adjuvanticity of flagellin. These observations indicate that the underlying mechanism remains incompletely understood. Here, we found that the native form of Salmonella typhimurium aa 89-96-mutant flagellin extracted from flagella retains some TLR5 recognition activity, indicating that aa 89-96 is the primary, but not the only site that imparts TLR5 activity. Additionally, this mutation impaired the production of IL-lp and IL-18. Using TLR5KO mice, we found that aa 89-96 is critical for the humoral adjuvant effect, but this effect was independent of TLR5 activation triggered by this region of flageUin. In summary, our findings suggest that aa 89-96 of flagellin is not only the crucial site responsible for TLR5 recognition, but is also important for humoral immune adjuvanticity through a TLR5-independent pathway.

关键词： adjuvanticity amino acids 89-96 Salmonella typhimurium flagellin TLR5 activity

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Proteomic Profiling of Purified Rabies Virus Particles

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Virologica Sinica 2020年第2期35卷 143-155页

作者： Yan Zhang Yuyang Wang Ye Feng Zhongzhong Tu Zhiyong Lou Changchun Tu key laboratory of Jilin Province for zoonosis prevention and control Institute of Military Veterinary MedicineAcademy of Military Medical SciencesAcademy of Military SciencesChangchun 130122China School of Medicine and collaborative innovation center of Biotherapy Tsinghua UniversityBeijing 100084China jiangsu co-innovation Centre for prevention and control of important animal infectious diseases and zoonoses Yangzhou 225009China

While host proteins incorporated into virions during viral budding from infected cell are known to play essential roles in multiple process of the life cycle of progeny virus,these characteristics have been largely neglected in studies on rabies virus(RABV).Here,we purified the RABV virions with good purity and integrity,and analyzed their proteome by nano LC–MS/MS,followed by the confirmation with immunoblot and immuno-electronic *** addition to the 5 viral proteins,49 cellular proteins were reproducibly identified to be incorporated into matured RABV *** annotation suggested that 24 of them were likely involved in virus ***,cryo-EM was employed to observe the purified RABV virions,generating high-resolution pictures of the bullet-shaped virion structure of *** study has provided new insights into the host proteins composition in RABV virion and shed the light for further investigation on molecular mechanisms of RABV infection,as well as the discovery of new anti-RABV therapeutics.

关键词： Rabies virus(RABV) Purification Virion structure Virion proteome

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Amino acids 89–96 of Salmonella flagellin: a key site for its adjuvant effect independent of the TLR5 signaling pathway

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Cellular & Molecular Immunology 2017年第12期14卷 1023-1025页

作者： Xilong Kang Zhiming Pan Xinan Jiao jiangsu key laboratory of zoonosis YangzhouUniversityYangzhouJiangsu 225009China jiangsu co-innovation center for the prevention and control of important animal infectious diseases and zoonoses Yangzhou UniversityYangzhouJiangsu 225009China key laboratory of prevention and control of Biological Hazard Factors(animal Origin)for Agrifood Safety and Quality MOAYangzhou UniversityYangzhouJiangsu 225009China Joint International Research laboratory of Agriculture and Agri-product Safety of the Ministry of Education YangzhouJiangsu 225009China

Flagellin,the main component of bacterial flagella,is a well-known pathogen-associated molecular pattern(PAMP)that can be recognized by the cell surface-localized Toll-like receptor 5(TLR5)and the cytosolic NOD-like receptor protein 4(NLRC4)inflammasome receptor NAIP5/6.1 Hosts can detect flagellated bacteria using the dual sensors,TLR5 and NAIP5/6,to recognize flagellin,and these sensors then initiate innate immune responses to protect the host from bacterial *** by extracellular TLR5 can trigger the MyD88-dependent signaling pathway and activate nuclear factorκB,leading to the upregulation of several proinflammatory cytokines.

关键词： effect TLR5 initiate

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Evaluating the efficacy of an attenuated Streptococcus equi ssp. zooepidemicus vaccine produced by multi-gene deletion in pathogenicity island SeseCisland_4

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Journal of Integrative Agriculture 2019年第5期18卷 1093-1102页

作者： MA Fang WANG Guang-yu ZHOU Hong MA Zhe LIN Hui-xing FAN Hong-jie MOE Joint International Research laboratory of animal Health and Food Safety College of Veterinary Medicine Nanjing Agricultural University National center of Meat Quality and Safety control Nanjing Agricultural University jiangsu co-innovation center for prevention and control of important animal infectious diseases and zoonoses Yangzhou University

Streptococcus equi ssp. zooepidemicus(SEZ) is a pathogen associated with a wild range of animal species. Frequent outbreaks have occurred in recent years in pigs, horses, goats and dogs which is liable to infect humans. There is a lack of efficient vaccines against this disease and the occurrence of antibiotic resistance may render drug therapies ineffective. In this study, gene deletion mutant(ΔSEZ) in pathogenicity islands SeseCisland_4 was constructed. The mutant ΔSEZ had a 52-fold decrease in 50% lethal dose(LD_(50)) and had less capacity to adhere epithelial cells. importantly, immunization of mice with attenuated vaccine ΔSEZ at the dose of 10~2 colony-forming units(CFU) mL^(–1) elicited a significant humoral antibody response, with an antibody titer of 1:12 800. Therefore, 10~2 CFU mL^(–1) might be used as the appropriate immune dose for the attenuated vaccine ΔSEZ, which provided mice with efficient protection against virulent SEZ. In addition, the hyperimmune sera against 10~2 CFU m L–1 attenuated vaccine ΔSEZ could confer significant protection against virulent SEZ infection in the passive immunization experiment and exhibited efficient bactericidal activity in the whole blood assay. Meanwhile, no viable bacteria was detected in blood when mice were immunized with ΔSEZ at the dose of 10~2 CFU mL^(–1) via hypodermic injection. Thereafter, the mutant ΔSEZ at the dose of 10~2 CFU mL^(–1) could confer significant protection in mice and had less negative effects on host, which could be an effective attenuated vaccine candidate for the prevention of SEZ.

关键词： Streptococcus equi ssp.zooepidemicus zoonosis attenuated vaccine immune dose

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Epizootiological surveillance of porcine circoviruses in free-ranging wild boars in China

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Virologica Sinica 2023年第5期38卷 663-670页

作者： Wenjie Gong Haiying Du Tong Wang Heting Sun Peng Peng Siyuan Qin Haidong Geng Zheng Zeng Wangwang Liang Hongquan Ling Changchun Tu Zhongzhong Tu State key laboratory for Diagnosis and Treatment of Severe Zoonotic infectious diseases Key Laboratory for Zoonosis Research of the Ministry of EducationInstitute of Zoonosisand College of Veterinary MedicineJilin UniversityChangchun130062China Changchun Veterinary Research Institute Chinese Academy of Agricultural SciencesChangchun130122China jiangsu co-innovation center for the prevention and control of important animal infectious Disease and zoonoses Yangzhou UniversityYangzhou225009China Biological Disaster control and prevention center National Forestry and Grassland AdministrationShenyang110034China Chongqing animal Disease prevention and control center Chongqing401120China

Four species of porcine circoviruses(PCV1–4)have been reported to circulate in Chinese domestic pigs,while the epizootiology of these viruses in free-ranging wild boars in China remains *** this study,tissue and serum samples collected from diseased or apparently healthy wild boars between 2018 and 2020 in 19 regions of China were tested for the prevalence of PCV1–4 *** rates of PCV1,PCV2,and PCV3 DNA in the tissue samples of Chinese wild boars were 1.6%(4/247),58.3%(144/247),and 10.9%(27/247)respectively,with none positive for *** analysis of viral genome showed that the four PCV1 strains distributed in Hunan and Inner Mongolia shared 97.5%–99.6%sequence identity with global distributed reference *** of the ORF2 gene sequences showed that 80 PCV2 strains widely distributed in 18 regions shared 79.5%–100%sequence identity with reference strains from domestic pigs and wild boars,and were grouped into PCV2a(7),PCV2b(31)and PCV2d(42).For PCV3,17 sequenced strains shared 97.2%–100%nucleotide identity at the genomic level and could be divided into PCV3a(3),PCV3b(2)and PCV3c(12)based on the phylogeny of ORF2 gene *** data revealed antibody positive rates against PCV1 and PCV2 of 11.4%(19/167)and 53.9%(90/167)*** data obtained in this study improved our understanding about the epidemiological situations of PCVs infection in free-ranging wild boars in China and will be valuable for the prevention and control of diseases caused by PCVs infection.

关键词： Porcine circoviruses(PCVs) Epizootiology Wild boar China

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Advances in pathogenesis of Streptococcus suis serotype 2

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Journal of Integrative Agriculture 2017年第12期16卷 2834-2847页

作者： FAN Hong-jie MOE Joint International Research laboratory of animal Health and Food Safety College of Veterinary Medicine Nanjing Agricultural University Nanjing 210095 P.R.China jiangsu co-innovation center for prevention and control of important animal infectious diseases and zoonoses Yangzhou 225009 P.R.China

Streptococcus suis is one of the major pathogens of swine streptococcosis. Among them, the strongest virulence and highest rate of clinical isolation serotype is S. suis serotype 2（SS2）. Moreover, SS2 is also an important zoonosis pathogen, which caused severe public health issues in China. It has been reported that SS2 has several virulence factors, including muramidase released protein, extracellular factors, capsule, fibronectin-binding protein, enolase, hemolysin, small RNA, biofilm, two-component regulatory systems, STK/STP, etc., whose functions involved in adhesion, anti-phagocytosis, inflammatory pathway activation, invasion, etc. Actually, SS2 has developed a variety of ways to escape from host immune system during evolution. In particularly, capsule could resist phagocytosis through inhibiting sphingosine dependent immune cell recognition, which plays an important role in escaping host inflammation response; moreover, superoxide dismutase encoding by sod A enables SS2 escaping reactive oxygen species（ROS） in host immune cells; besides, binding complement factor h with Fhb could suppress the activation of complement alternative pathway and bactericidal effect. And SS2 could also hinder the formation of neutrophil extracellular traps（NETs） to avoid trapping by swine neutrophils, while host immune globulin could be degraded by Ig A1 hydrolase and Ig M protease. In addition, SS2 could escape host immune defense with the help of multiple transcriptional factors and micro-RNA. So far, the pathogenesis of meningitis, arthritis caused by SS2 infection, is still unclear, and the virulence regulatory mechanism of phosphorylation, micro-RNA need to be further clarified. importantly, the study of interaction mechanism of pathogen and host contribute to further demonstration the pathogenesis of SS2.

关键词： Advances in pathogenesis of Streptococcus suis serotype 2

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Human fused NKG2D-IL-15 protein controls xenografted human gastric cancer through the recruitment and activation of NK cells

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Cellular & Molecular Immunology 2017年第3期14卷 293-307页

作者： Yan Chen Bei Chen Ti Yang Weiming Xiao Li Qian Yanbing Ding Mingchun Ji Xiaoqun Ge Weijuan Gong Department of Immunology School of Medicine Yangzhou University Yangzhou 225001 People's Republic of China Department of Pharmacology School of Medicine Yangzhou University Yangzhou 225001 People's Republic of China Department of Gastroenterology The Second Affiliated Hospital Yangzhou University Yangzhou 225001 People's Republic of China jiangsu key laboratory of Integrated Traditional Chinese and Western Medicine for prevention and Treatment of Senile diseases Yangzhou University Yangzhou 225009 People's Republic of China jiangsu key laboratory of zoonosis Yangzhou University Yangzhou 225009 People's Republic of China jiangsu co-innovation center for prevention and control of important animal infectious diseases and zoonoses Yangzhou 225009 People's Republic of China

Interleukin （IL）-15 plays an important role in natural killer （NK） and CD8＋ T-cell proliferation and function and is more effective than IL-2 for tumor immunotherapy. The trans-presentation of IL-15 by neighboring cells is more effective for NK cell activation than its soluble IL-15. In this study, the fusion protein dsNKG2D-IL-15, which consisted of two identical extracellular domains of human NKG2D coupled to human IL-15 via a linker, was engineered in Escherichia coll. DsNKG2D-IL- 15 could efficiently bind to major histocompatibility complex class I chain-related protein A （MICA） of human tumor cells with the two NKG2D domains and trans-present IL-15 to NK or CD8＋ T cells. We transplanted human gastric cancer （SGC-7901） cells into nude mice and mouse melanoma cells with ectopic expression of MICA （B 16BL6-MICA） into C57BL/6 mice. Then, we studied the anti-tumor effects mediated by dsNKG2D-IL-15 in the two xenografted tumor models. Human dsNKG2D-IL-15 exhibited higher efficiency than IL-15 in suppressing gastric cancer growth. Exogenous human dsNKG2D-IL-15 was centrally distributed in the mouse tumor tissues based on in vivo live imaging. The frequencies of human CD56＋ cells infiltrated into the tumor tissues following the injection of peripheral blood mononuclear cells into nude mice bearing human gastric cancer were significantly increased by human dsNKG2D-I L- 15 treatment. Human dsNKG2 D-I L- 15 also delayed the growth of transplanted melanoma （B 16BL6- MICA） by activating and recruiting mouse NK and CD8＋ T cells. The anti-melanoma effect of human dsNKG2D-IL-15 in C57BL/6 mice was mostly decreased by the in vivo depletion of mouse NK cells. These data highlight the potential use of human dsNKG2D-IL-15 for tumor therapy.

关键词： fusion protein IL-15 NK cells NKG2D tumor

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Neutralization activity of influenza A virus humanized antibodies against new subtypes of influenza viruses

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Biosafety and Health 2019年第3期1卷 159-161页

作者： Jing Liu Tiecheng Wang Ying Xie Yuanguo Li Jian He Xinghai Zhang Weiyang Sun Na Feng Chuan Qin Yuwei Gao Xianzhu Xia comparative Medicine center Peking Union Medical College(PUMC)and Institute of Laboratory Animal SciencesChinese Academy of Medical Sciences(CAMS)Beijing 100021China key laboratory of Jilin Province for zoonosis Institute of Military VeterinaryAcademy of Military Medical SciencesChangchun 130122China jiangsu co-innovation center for prevention and control of important animal infectious diseases and zoonoses Nanjing 225009China

Antibodies are ideal for controlling the influenza A virus,but their effect on newly emerging strains is ***,we assessed the neutralization activity of the humanized monoclonal antibodies(mAbs)F10,H98 and H40 against circulating influenza viruses(H5N1,H1N1,H3N2 and H7N7 and new subtypes viruses H5N6 and H7N9).The results showed that all the three humanized mAbs(F10,H98 and H40)displayed different degrees of virus neutralization activities when encountered with different subtypes of influenza ***,the humanized monoclonal antibody F10 produced higher and broader neutralization titers(range 25–1.56μg/ml)than those of the other two humanized mAbs(H98(range 50–3.12μg/ml),H40(range 50–5.56μg/ml))to against the viruses H5N1,H1N1,H3N2,H7N7,H5N6 and *** mAb may represent a new class of heterosubtypic neutralizing humanized mAb that could replace vaccines and chemical drugs.

关键词： Influenza A virus humanized antibodies Neutralization activity New subtypes of influenza viruses

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