Jialiang Pan

Nanjing Agricultural University

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Jianjun Dai

Nanjing Agricultural University

Mengting Liu

Nanjing Agricultural University

Fang Tang

Chinese University of Hong Kong

Youli Yu

Nanjing Agricultural University

Hongsheng Wang

Chinese Academy of Medical Sciences & Peking Union Medical College

Haiqin Jiang

Chinese Academy of Medical Sciences & Peking Union Medical College

Jingshu Xiong

Chinese Academy of Medical Sciences & Peking Union Medical College

Feng Xue

Shanghai Jiao Tong University

Lei Tao

ZheJiang Institute For Food and Drug Control

Lei Tao

Fudan University

Cooperative Institutions

Nanjing Agricultural University

Ministère de l'Agriculture et de la Souveraineté alimentaire

Peking University

Chinese Academy of Medical Sciences & Peking Union Medical College

Département Santé Animale

Fudan University

Peking University First Hospital

Wenzhou Medical University

Institute of Dermatology

Peking Union Medical College Hospital

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Guanylate binding protein 2b contributes to M1 macrophage polarization during Mycobacterium bovis infection

Research Square (Research Square) (2022)

Youli Yu Mengting Liu Jialiang Pan Lei Tao Feng Xue

Abstract Accumulating evidence has demonstrated that the macrophage phenotype switch from M0 to M1 is critical in the clearance of intracellular mycobacteria during Mycobacterium bovis infection. In this study, the entire gene expression profiles of M1 bone marrow-derived macrophages (BMDMs) induced by interferon-gamma and lipopolysaccharide, M2 BMDMs induced by interleukin (IL) - 4 and IL-13, M0 BMDMs, and M0 BMDMs after 6 h of infection with M. bovis were subjected to bioinformatics analysis. Forty-eight candidate genes associated with M. bovis infection and involved in M1 polarized macrophages were screened. Thirteen hub genes were subsequently identified based on connectivity degree analysis and multiple external validations. Among these hub genes, Guanylate binding protein 2b (GBP2b) was selected as a key gene for further investigation. In vivo validation results showed significant expression of GBP2b in the tissues of M. bovis -infected mice. For in vitro validation, the RAW264.7 cell line was transfected with GBP2b-specific small interfering RNA and GBP2b plasmid expression vector. Cellular experimental studies confirm that GBP2b was a novel regulatory molecule that promotes M1 macrophage polarization during M. bovis infection and was also an antimicrobial protein that promotes intracellular bacterial clearance. Furthermore, gene co-expression analysis and further experiments showed that GBP2b regulates M1 polarization through activation of Toll-like receptor 4 and nuclear factor-κBp65 protein. These findings expand the understanding of GBP2b for the regulation of inflammatory responses and suggest that GBP2b may be a potential target for the treatment of diseases caused by M. bovis.

Macrophage polarization

10.21203/rs.3.rs-1631961/v1

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Guanylate-binding protein 2b regulates the AMPK/mTOR/ULK1 signalling pathway to induce autophagy during Mycobacterium bovis infection

Figshare (2022)

Youli Yu Jialiang Pan Mengting Liu Haiqin Jiang Jingshu Xiong

Mycobacterium bovis is a major cause of animal tuberculosis that is also highly dangerous to human health. Autophagic isolation and degradation of intracellular pathogens are employed by host cells as primary innate immune defence mechanisms to control intercellular M. bovis infection. In this study, RNA-Seq technology was used to obtain the total mRNA from bone marrow-derived macrophages (BMDMs) infected with M. bovis at 6 and 24 h after infection. One of the differential genes, GBP2b, was also investigated. Analysis of the significant pathway involved in GBP2b-coexpressed mRNA demonstrated that GBP2b was associated with autophagy and autophagy-related mammalian target of rapamycin (mTOR) signalling and AMP-activated protein kinase (AMPK) signalling. The results of in vivo and in vitro experiments showed significant up-regulation of GBP2b during M. bovis infection. For in vitro validation, small interfering RNA-GBP2b plasmids were transfected into BMDMs and RAW264.7 cells lines to down-regulate the expression of GBP2b. The results showed that the down-regulation of GBP2b impaired autophagy via the AMPK/mTOR/ULK1 pathway. Further studies revealed that the activation of AMPK signalling was essential for the regulation of autophagy during M. bovis infection, and the down-regulation of GBP2b promoted the intracellular survival of M. bovis. These findings were validated on two types of macrophages, which extended the knowledge about the involvement of GBP2b in the autophagic process. Based on these observations, GBP2b should be developed as a promising molecular target for intervening on host–pathogen interactions to develop novel therapeutic strategies to control M. bovis infections in humans and animals.

ULK1

Signalling

10.6084/m9.figshare.19732230.v1

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Combined Therapy of Probiotic Microcapsules and Bomidin in Vibrio parahaemolyticus–Infected Rats

Life (2022)

Ting Zhou Mengting Liu Jialiang Pan Jianluan Ren Fang Tang

Background: With the discovery of more and more drug–resistant bacterial strains, there is an urgent need for safer and more effective alternative treatments. In this study, antibacterial peptides and probiotic microcapsules were combined to treat gastrointestinal inflammation caused by Vibrio parahaemolyticus infection. Methods: To improve the stability of probiotics in the gastrointestinal tract, two types of mixed natural anionic polysaccharides and chitosan were used as carriers to embed the probiotics. Taking Lacticaseibacillus casei CGMCC1.8727 microcapsules with good performance as the research object, the in vitro characteristics of the microcapsules were studied via acid resistance test and intestinal release test. The microcapsules were then tested for in vivo treatment in combination with the antibacterial peptide, bomidin, and the therapeutic effects were compared among microencapsulated probiotics, free probiotics, and probiotics in combination with bomidin. Results: Microencapsulation was successfully manufactured under suitable processing parameters, with the product particle size being 2.04 ± 0.2743 mm. Compared with free probiotics, microencapsulation significantly improved the activity and preservation stability of the probiotics under simulated gastrointestinal conditions. Microencapsulated probiotics showed better therapeutic effects than free probiotics in vivo. Microcapsules combined with antimicrobial peptides accelerated the elimination of bacteria in vivo. This study provides a reference for anti–inflammatory treatment, especially for the treatment of gastrointestinal diseases.

Lactobacillus casei

10.3390/life12111740

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Guanylate-Binding protein 2b regulates the AMPK/mTOR/ULK1 signalling pathway to induce autophagy during Mycobacterium bovis infection

Figshare (2022)

Youli Yu Jialiang Pan Mengting Liu Haiqin Jiang Jingshu Xiong

Autophagic isolation and degradation of intracellular pathogens are employed by host cells as primary innate immune defense mechanisms to control intercellular M. bovis infection. In this study, RNA-Seq technology was used to obtain the total mRNA from bone marrow-derived macrophages (BMDMs) infected with M. bovis at 6 and 24 h after infection. One of the differential genes, GBP2b, was also investigated. Analysis of the significant pathway involved in GBP2b-coexpressed mRNA demonstrated that GBP2b was associated with autophagy and autophagy-related mammalian target of rapamycin (mTOR) signaling and AMP-activated protein kinase (AMPK) signaling. The results of in vivo and in vitro experiments showed significant up-regulation of GBP2b during M. bovis infection. For in vitro validation, small interfering RNA-GBP2b plasmids were transfected into BMDMs and RAW264.7 cells lines to down-regulate the expression of GBP2b. The results showed that the down-regulation of GBP2b impaired autophagy via the AMPK/mTOR/ULK1 pathway, thereby promoting the intracellular survival of M. bovis. Further studies revealed that the activation of AMPK signaling was essential for the regulation of autophagy during M. bovis infection. These findings expand the understanding of how GBP2b regulates autophagy and suggest that GBP2b may be a potential target for the treatment of diseases caused by M. bovis.

ULK1

Signalling

10.6084/m9.figshare.19732230

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Guanylate-Binding protein 2b regulates the AMPK/mTOR/ULK1 signalling pathway to induce autophagy during Mycobacterium bovis infection

Virulence (2022)

Youli Yu Jialiang Pan Mengting Liu Haiqin Jiang Jingshu Xiong

Autophagic isolation and degradation of intracellular pathogens are employed by host cells as primary innate immune defense mechanisms to control intercellular M. bovis infection. In this study, RNA-Seq technology was used to obtain the total mRNA from bone marrow-derived macrophages (BMDMs) infected with M. bovis at 6 and 24 h after infection. One of the differential genes, GBP2b, was also investigated. Analysis of the significant pathway involved in GBP2b-coexpressed mRNA demonstrated that GBP2b was associated with autophagy and autophagy-related mammalian target of rapamycin (mTOR) signaling and AMP-activated protein kinase (AMPK) signaling. The results of in vivo and in vitro experiments showed significant up-regulation of GBP2b during M. bovis infection. For in vitro validation, small interfering RNA-GBP2b plasmids were transfected into BMDMs and RAW264.7 cells lines to down-regulate the expression of GBP2b. The results showed that the down-regulation of GBP2b impaired autophagy via the AMPK/mTOR/ULK1 pathway, thereby promoting the intracellular survival of M. bovis. Further studies revealed that the activation of AMPK signaling was essential for the regulation of autophagy during M. bovis infection. These findings expand the understanding of how GBP2b regulates autophagy and suggest that GBP2b may be a potential target for the treatment of diseases caused by M. bovis.

Intracellular parasite

10.1080/21505594.2022.2073024

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Citations (5)