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PLA_(2) mediates the Innate immune response in Asian corn borer, Ostrinia furnacalis

Insect Science 2022年第1期29卷 245-258页

作者： Jia-Yue Ji Zhao-Hua Yin Sha-Sha Zhang Dong-Xu Shen Chun-Ju An Department of Entomology College of Plant ProtectionChina Agricultural UniversityBeijing100193China Jiangsu Key Laboratory of Sericutural Biology and Biotechnology School of BiotechnologyJiangsu University of Science and TechnologyZhenjiangJiangsu212018China Key Laboratory of Silkworm and Mulberry Genetic Improvement Ministry of Agriculture and Rural AffairsSericultural Research InstituteChinese Academy of Agricultural SciencesZhenjiangJiangsu212018China

The eicosanoid signaling pathway mediates insect immune reactions to a wide range of stimuli. This pathway begins with the biosynthesis of arachidonic acid (AA) from the hydrolysis of phospholipids catalyzed by phospholipase A_(2) (PLA_(2)). We report here that the PLA_(2) inhibitor, dexamethasone (DEX), impaired the Innate immune response including nodulation, encapsulation, and melanization in Ostrinia furnacalis larvae, while AA partially reversed these effects of DEX. We cloned a full-length complementary DNA encoding a PLA_(2), designated as OfsPLA_(2), from O. furnacalis. The open reading frame of OfsPLA_(2) encodes a 195-amino acid residue protein with a 22-residue signal peptide. Sequence alignment analyses indicated that O. furnacalis PLA_(2) might be a Group III secretory PLA_(2). The highest transcript levels of OfsPLA_(2) were detected in the fat body, and its transcript levels increased dramatically after infection with Escherichia coli, Micrococcus luteus, or Beauveria bassiana. Recombinant OfsPLA_(2) significantly induced prophenoloxidase (PPO) activation in larval hemolymph in the presence of Ca^(2+) and encapsulation of agarose beads. Injection of recombinant OfsPLA_(2) into larvae resulted in increased transcript levels of attacin, defencin, and moricin-3 genes. Our results demonstrate the involvement of the eicosanoid signaling pathway in the Innate immune response of O. furnacalis larvae and provide new information about the roles of O. furnacalis secretory PLA_(2) in activating PPO and antimicrobial peptide production.

关键词： eicosanoid signal pathway Innate immune response Ostrinia furnacalis phospholipase A_(2)(PLA_(2))

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p62/SQSTM1 Participates in the Innate immune response of Macrophages Against Candida albicans Infection

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International Journal of Dermatology and Venereology 2022年第2期5卷 75-81页

作者： Yan-Zhi He Zhi-Min Duan Xu Chen Min Li Center for Global Health School of Public HealthNanjing Medical UniversityNanjingJiangsu 211166China Department of Dermatology Hospital for Skin Diseases(Institute of Dermatology)Chinese Academy of Medical Sciences and Peking Union Medical CollegeNanjingJiangsu 210042China Jiangsu Key Laboratory of Molecular Biology for Skin Diseases and STIs Hospital for Skin Diseases(Institute of Dermatology)Chinese Academy of Medical Sciences and Peking Union Medical CollegeNanjingJiangsu 210042China.

Objective: This study was designed to evaluate whether p62/SQSTM1 (hereafter referred to as p62) is involved in the immune response of macrophages against challenge byCandida albicans (C. albicans).Methods: We cultured bone marrow-derived macrophages (BMDMs) to investigate the immune response to challenge byC. albicans. The p62 gene was knocked down by transfection with p62 small interfering RNA (siRNA) in the p62 siRNA group. BMDMs transfected with nonsense siRNA served as the negative control (NC) group. These two groups of BMDMs were challenged withC. albicans in vitro. We detected p62 expression through quantitative reverse transcription PCR and western blotting. The phagocytosis ability of BMDMs was evaluated by flow cytometry and microscopic examination using an Olympus FV1000 laser scanning confocal microscope. Moreover, we determined the level of reactive oxygen species (ROS) in BMDMs. The mRNA levels of proinflammatory cytokines were determined by quantitative reverse transcription PCR.Results: After stimulation byC. albicans, the relative expression of p62 mRNA was increased in a dose-dependent manner, the relative expression of p62 and the ratio of BMDMs toC. albicans is 1.893 ± 0.2156 (1:1,P < 0.05), 2.873 ± 0.4787 (1:3,P < 0.05) and 3.556 ± 0.2892 (1:5,P < 0.01). The p62 protein level was also increased. After transfection with p62 siRNA, the mRNA and protein levels of p62 were significantly decreased in BMDMs (P < 0.05). After 0.5, 1 and 2 hours of co-culture of BMDMs withC. albicans, flow cytometry showed that the phagocytosis rates ofC. albicans by BMDMs were significantly lower in the p62 siRNA group than in the NC group (39.70 ± 1.69%vs. 55.23 ± 0.72%, 46.70 ± 0.89%vs. 60.80 ± 1.78%, 51.90 ± 0.98%vs. 64.43 ± 2.0%, respectively, allP < 0.05). Consistent results were seen in the production of ROS (4269 ± 392.6vs. 13426 ± 1859.7, 4967 ± 721.2vs. 13687 ± 2611.2, 7647 ± 1950.0vs. 17719 ± 1814.2, respectively, allP < 0.05). The ROS levels were higher in BMDMs of the NC group than in BMDMs transfected

关键词： p62/SQSTM1 Candida albicans macrophage Innate immune response phagocytosis

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The antimicrobial peptide Brevinin-2ISb enhances the Innate immune response against methicillinresistant Staphylococcus aureus by activating DAF-2/DAF-16 signaling in Caenorhabditis elegans,as determined by in vivo imaging

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Journal of Bio-X Research 2020年第4期3卷 205-218页

作者： Hui Xie Xu Nie Yonghua Zhan Qi Zeng Xueli Chen Dan Chen School of Life Science and Technology Xidian UniversityXi’anShaanxi ProvinceChina

Objective::In order to study the important role and molecular mechanism of Brevinin-2 family antimicrobial peptide Brevinin-2ISb in methicillin-resistant Staphylococcus aureus(MRSA)infection of Caenorhabditis(C.)elegans,and to find the optimal therapeutic concentration of Brevinin-2ISb.Methods::By using a C.elegans model and MRSA infection modelto study the therapeutic effect of different concentrations of Brevinin-2ISb on C.elegans.Real-time PCR was used for investigating the effect of Brevinin-2ISb on the downstream gene expression of DAF-2/DAF-16 innate immune pathway and the major virulence factor gene expression of MRSA.With protein activity tests to study the inhibitory effect of Brevinin-2ISb on MRSA virulence factor protein activity.Finally,laser confocal imaging was carried out to observe real-time expression and distribution of downstream antimicrobial proteins to further prove the effect of Brevinin-2ISb on the activation of DAF-2/DAF-16 pathway by in vivo imaging.All animal study procedures were approved by the Academic Committee at Xidian University and Xi’an Jiaotong University Animal Care and Use Committee,China(approval No.JGC201207)on July 15,2017.Results::Host immunity was largely enhanced by Brevinin-2ISb,and the expression of staphylococcal enterotoxin genes,as well as virulence factors,was suppressed by Brevinin-2ISb.Indeed,the expression of many C.elegans innate immune genes,including lys-7,spp-1,K05D8.5 and C29F3.7,was induced by Brevinin-2ISb.In particular,robust,sustained expression of the antibacterial gene lys-7 was observed after Brevinin-2ISb treatment,resulting in increased protein levels.These effects correlated with a reduction in the MRSA-mediated death of the C.elegans host.Low concentrations of Brevinin-2ISb exhibited very low hemolytic activity,and may play a positive role in host innate immunity.Specifically,activation of the DAF-2/DAF-16 pathway appears to be essential for immune activation in C.elegans treated with Brevinin-2ISb.Based on the evolutionary conservation of inna

关键词： brevinin-2ISb caenorhabditis elegans DAF-2/DAF-16 Innate immune response MRSA

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Nucleotide oligomerization domain 2 contributes to the Innate immune response in THCE cells stimulated by Aspergillus fumigatus conidia

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International Journal of Ophthalmology(English edition) 2012年第4期5卷 409-414页

作者： Zheng-Jie Xu Gui-Qiu Zhao Qing Wang Cheng-Ye Che Nan Jiang Li-Ting Hu and Qiang Xu Department of Ophthalmology the Affiliated Hospital of Qingdao University Medical College Qingdao 266003 Shandong Province China

AIM: To investigate the expression of nucleotide oligomerization domain 2 (NOD2) in the immortalized human corneal epithelial cell line (THCE), and its role in the Innate immune response triggered by inactive Aspergillus fumigatus (Af) conidia. METHODS: The normal THCE cells were investigated as controls. After incubation with inactive Af conidia for 0.5, 2, 4, 6, and 8 hours, THCE cells were harvested, mRNA expression of NOD2 and receptor interacting protein 2 (RIP2) was detected by RT-PCR. Intracellular proteins including NOD2, NF-kappa B and proinflammatory cytokines such as TNF-alpha, IL-8, IL-6 in the cell supernatant were analyzed by ELISA. RESULTS: Our data indicate that NOD2 expressed in the normal THCE cells. After triggered by the inactive Af conidia, the expression of NOD2, RIP2 mRNA and the secretion of NOD2, NF-kappa B, TNF-alpha, IL-8, IL-6 both increased in a time-depended manner, and reached the peak point at 4, 6, 6, 4, 6, 6, 4 hours, respectively. And after pretreated with NOD2 neutralizing antibody, the expression of RIP2, NF-kappa B, TNF-alpha, IL-8 both decreased dramatically at the peak point, while the secretion of IL-6 changed little. CONCLUSION: The results of this study suggest that NOD2 exists and expresses in the THCE cells, and contributes to the Innate immune responses triggered by inactive Afconidia by induction of proinflammatory cytokines such as TNF-alpha and IL-8 through the NF-kappa B pathway.

关键词： nucleotide oligomerization domain 2 corneal epithelial cell Aspergillus fumigatus Innate immune response

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The SUMOylation of TAB2 mediated by TRIM60 inhibits MAPK/NF-κB activation and the Innate immune response

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Cellular & Molecular Immunology 2021年第8期18卷 1981-1994页

作者： Zhiwen Gu Xueying Chen Wenyong Yang Yu Qi Hui Yu Xiaomeng Wang Yanqiu Gong Qianqian Chen Bo Zhong Lunzhi Dai Shiqian Qi Zhiqiang Zhang Huiyuan Zhang Hongbo Hu Department of Rheumatology and Immunology National Clinical Research Center for GeriatricsState Key Laboratory of BiotherapyWest China HospitalSichuan Universityand Collaborative Innovation Center of BiotherapyChengdu610041China Department of Virology College of Life SciencesDepartment of ImmunologyMedical Research InstituteWuhan UniversityWuhan430072China Department of General Practice and Lab of PTM State Key Laboratory of BiotherapyWest China HospitalSichuan Universityand Collaborative Innovation Center of BiotherapyChengdu610041China Department of Urology State Key Laboratory of BiotherapyWest China HospitalSichuan Universityand Collaborative Innovation Center of BiotherapyChengdu610041China Immunobiology and Transplant Science Center Houston Methodist HospitalHoustonTXUSA77030

Activation of the TAK1 signalosome is crucial for mediating the Innate immune response to pathogen invasion and is regulated by multiple layers of posttranslational modifications,including ubiquitination,SUMOylation,and phosphorylation;however,the underlying molecular mechanism is not fully understood.In this study,TRIM60 negatively regulated the formation and activation of the TAK1 signalosome.Deficiency of TRIM60 in macrophages led to enhanced MAPK and NF-κB activation,accompanied by elevated levels of proinflammatory cytokines but not IFN-I.Immunoprecipitation-mass spectrometry assays identified TAB2 as the target of TRIM60 for SUMOylation rather than ubiquitination,resulting in impaired formation of the TRAF6/TAB2/TAK1 complex and downstream MAPK and NF-κB pathways.The SUMOylation sites of TAB2 mediated by TRIM60 were identified as K329 and K562;substitution of these lysines with arginines abolished the SUMOylation of TAB2.In vivo experiments showed that TRIM60-deficient mice showed an elevated immune response to LPS-induced septic shock and L.monocytogenes infection.Our data reveal that SUMOylation of TAB2 mediated by TRIM60 is a novel mechanism for regulating the Innate immune response,potentially paving the way for a new strategy to control antibacterial immune responses.

关键词： SUMOylation Innate immune response TRAM60 TAB2

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Mycobacterium tuberculosis Mce2E suppresses the macrophage Innate immune response and promotes epithelial cell proliferation

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Cellular & Molecular Immunology 2019年第4期16卷 380-391页

作者： Lihua Qiang Jing Wang Yong Zhang Pupu Ge Qiyao Chai Bingxi Li Yi Shi Lingqiang Zhang George Fu Gao Cui Hua Liu CAS Key Laboratory of Pathogenic Microbiology and Immunology Institute of Microbiology Chinese Academy of Sciences100101 BeijingChina Institute of Health Sciences Anhui University230601 Hefei China Savaid Medical School University of Chinese Academy of Sciences 101408 Beijing China State Key Laboratory of Proteomics Beijing Proteome Research CenterNational Center of Protein Sciences BeijingBeijing Institute of Lifeomics100850 Beijing China

The intracellular pathogen Mycobacterium tuberculosis(Mtb)can survive in the host and cause disease by interfering with a variety of cellular functions.The mammalian cell entry 2(mce2)operon of Mtb has been shown to contribute to tuberculosis pathogenicity.However,little is known about the regulatory roles of Mtb Mce2 family proteins towards host cellular functions.Here we show that the Mce2 family protein Mce2E suppressed the macrophage Innate immune response and promoted epithelial cell proliferation.Mce2E inhibited activation of the extracellular signal-regulated kinase(ERK)and Jun N-terminal kinase(JNK)mitogen-activated protein kinase(MAPK)signaling pathways in a non-canonical D motif(a MAPK-docking motif)-dependent manner,leading to reduced expression of TNF and IL-6 in macrophages.Furthermore,Mce2E promoted proliferation of human lung epithelium-derived lung adenoma A549 cells by inhibiting K48-linked polyubiquitination of eEF1A1 in aβstrand region-dependent manner.In summary,Mce2E is a novel multifunctional Mtb virulence factor that regulates host cellular functions in a niche-dependent manner.Our data suggest a potential novel target for TB therapy.

关键词： Mycobacterium tuberculosis Mce2E Innate immune response cell proliferation

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ZDHHC11 modulates Innate immune response to DNA virus by mediating MITA–IRF3 association

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Cellular & Molecular Immunology 2018年第10期15卷 907-916页

作者： Ying Liu Qian Zhou Li Zhong Heng Lin Ming-Ming Hu Yan Zhou Hong-Bing Shu Shu Li College of Life Sciences Medical Research InstituteWuhan UniversityWuhanHubei 430072China

MITA is a central adaptor in Innate immune responses to DNA viruses.The mechanisms responsible for recruitment of downstream kinase TBK1 and the transcription factor IRF3 to MITA remains enigmatic.Here we identified ZDHHC11,a member of DHHC palmitoyl transferase family,as a positive regulator of DNA virus-triggered signaling.Overexpression of ZDHHC11 activated the IFN-βpromoter,while ZDHHC11-deficiency specifically impaired DNA virus HSV-1-induced transcription of downstream antiviral genes.Zdhhc11^(−/−)mice exhibited lower serum cytokine levels and higher lethality after HSV-1 infection.Mechanistically,ZDHHC11 facilitated the optimal recruitment of IRF3 to MITA.Our findings support an important role for ZDHHC11 in mediating MITA-dependent Innate immune responses against DNA viruses.

关键词： ZDHHC11 MITA IRF3 Innate immune response

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Peptidoglycan recognition protein 2, a Potential Target for Regulating Innate immune response in Hepatocytes

Peptidoglycan recognition protein 2, a Potential Target for ...

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第十四届全国免疫学学术大会

作者： Ming Shi School of Life Science and Technology Harbin Institute of Technology

Pattern recognition receptors(PRRs) function as the initial factors of the Innate immune response, which are closely associated with remodeling of the immune microenvironment, antitumor and antivirus immune response. Peptidoglycan recognition protein 2(PGLYRP2) is a pattern recognition receptor that is specifically expressed in liver and implicated in the regulation of innate immunity and immunosurveillance. However, the role of hepatic PGLYRP2 in modulating immune responses remains to be investigated. Recently, we have demonstrated that PGLYRP2 was down-regulated in HCC, which was linked with poor prognosis in patients. PGLYRP2 overexpression in HCC cells significantly enhanced antitumor immune responses in immune-competent mice and elevated immune response rates of peripheral blood mononuclear cells against HCC. Mechanistically, DNA methyltransferase 3A–mediated promoter hypermethylation was responsible for the down-regulation of PGLYRP2 in HCC. PGLYRP2 promoted production of chemokine(C-C motif) ligand 5(CCL5) in HCC through binding to the CCL5 promoter, which contributed to the enhanced antitumor immunity. Furthermore, PGLYRP2 significantly improved the survival rates and prognosis of hepatitis virus-positive patients with liver cancer. And PGLYRP2 could inhibit HBV gene expression and viral replication in vitro. Our data showed that PGLYRP2-mediated antiviral Innate immune response through interacting with HBV capsid and Core promoter. Moreover, the interactions among PGLYRP2, HBV DNA and capsid facilitate the recognition of hepatitis virus and regulation of hepatic immune microenvironment. The hepatocyte model of chronic HBV infection and mouse models of acute HBV infection further indicate that the PGLYRP2-mediated innate immune signaling potentially promotes the immunotherapy against chronic hepatitis B virus infection. These findings provide a new direction and a novel target for HCC and hepatitis virus immunotherapy.

关键词： Pattern recognition receptors signal Antivirus Innate immune response PGLYRP2

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MLL5 suppresses antiviral Innate immune response by facilitating STUB1-mediated RIG-I degradation

MLL5 suppresses antiviral innate immune response by facilita...

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第十二届全国免疫学学术大会

作者： Zhou Peipei Zhang Wei Ding Xiaodan Wan Xiaoling Liu Lulu Yuan Xiujie Hui Xinhui Meng Guangxun Xiao Hui Li Bin Zhong Jin Hou Fajian Deng Lihwen Zhang Yan Institut Pasteur of Shanghai Chinese Academy of Sciences Institute of Immunology Shanghai Jiaotong University Institute of Biochemistry and Cell Biology Chinese Academy of Sciences National University of Singapore

Trithorax group protein MLL5 is an important epigenetic modifier that controls cell cycle progression,chromatin architecture maintenance and hematopoiesis.However,whether MLL5 has a role in innate antiviral immunity is largely unknown.Here,we found that MLL5 suppresses RIG-I-mediated anti-viral immune response.M115-deficient mice infected with vesicular stomatitis virus exhibited enhanced anti-viral innate immunity,and reduced morbidity and viral load.Mechanistically,a small fraction of MLL5 protein located in the cell cytoplasm interacted with both RIG-I and its E3 ubiquitin ligase STUBl,which promotes K48-linked polyubiquitination and proteasomal degradation of RIG-I.Deficiency of MLL5 attenuated the association between RIG-I and STUBl and reduced K48-linked polyubiquitination and accumulation of RIG-I protein in cells.In addition,upon virus infection,nuclear MLL5 protein translocated from the nucleus to the cytoplasm to induce STUB1-mediated degradation of RIG-I.Our study uncovers a previously unrecognized role for MLL5 in antiviral Innate immune responses and provides a new target for controlling viral infection.

关键词： Innate immune response MLL5 RIG-I STUB1

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Differences of Innate immune response to ALV-J infection between the slow feathering and the fast feathering chickens

Differences of innate immune response to ALV-J infection bet...

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第十一届全国免疫学学术大会

作者： Li Yuanfang South China Agricultural University

Feather-sexable lines is widely used in poultry industry, but the phenomenon show that the slow feathering are more sensitive to avian leucosis and more difficult to purify than fast feather. The slow feathering(SF) gene linked with the ev21 gene which is an endogenous retrovirus(EV) incorporation may cause different immune effect to avian leukemia virus(ALV) infection. In this study, we intend to explore the differences of the Innate immune response to ALV-infected between the slow feathering and the fast feathering.We measured the expression of the virus, innate immune molecule interferon( IFN-α), interleukin-10(IL10) and IL12, tumor necrosis factor(TNF-α) and antiviral genes(Mx, PKR, and OAS) between the slow feathering and the fast feathering.In our data, the virus replication level of the infected-slow feathering group began to increase earlier than the infectedfast feathering and the replication quantity is lower than the fast feathering whether in spleen or lymphocyte. The protein levels of IL-10, IL-12, TNF-α and IFN-α of the slow feathering infected group increase rapidly at 11 d.p.i. and shown no significant differences with the control group, the cytokine levels of the infected group increase sharply at 13 d.p.i.(p innate immunity system of the slow feathering were broken earlier than the fast feathering and the slow feathering chickens have immunologic tolerance compared with the fast feathering chickens.

关键词： ALV-J ev21 cytokines Innate immune response slow feathering and fast feathering

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