Cell Host & Microbe
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2024
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IF 20.6
Xin Wang,Huihui Kong,Bingxin Chu,Qian Yang,Chaohui Lin,Rui Liu,Changxu Chen,Yang Gao,Guojun Wang,Dayan Wang,Chen Qin,Xiaohua Ye,Lifei Yu,Xiangfei Xu,Jie Jin,Ren Sun,Hualan Chen,Xudong Wu,Zeli Zhang
Identifying broadly reactive B precursor cells and conserved epitopes is crucial for developing a universal flu vaccine. In this study, using influenza neuraminidase (NA) mutant probes, we find that human pre-existing NA-specific memory B cells (MBCs) account for ∼0.25% of total MBCs, which are heterogeneous and dominated by class-unswitched MBCs. In addition, we identify three NA broad-inhibition monoclonal antibodies (mAbs) (BImAbs) that block the activity of NA derived from different influenza strains, including the recent cow H5N1. The cryoelectron microscopy (cryo-EM) structure shows that the BImAb targets the conserved NA enzymatic pocket and a separate epitope in the neighboring NA monomer. Furthermore, the NA BImAbs protect mice from the lethal challenge of the human pandemic H1N1 and H5N1. Our work demonstrates that the NA broad-inhibition precursor MBCs exist in healthy adults and could be targeted by the NA-based universal flu vaccine.
研究领域:
免疫学
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病毒学
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结构生物学
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疫苗开发
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DOI
PubMed
高影响因子
ACS Applied Nano Materials
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2024
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IF 5.3
Chu-shu Zhu,Siyue Xiong,Chengxuan Wang,Peitao Dong,Xuezhong Wu
Accurate and convenient diagnosis of the influenza virus is of critical importance to prevent the spread of flu infections and guide therapy and treatment. This work reports the fabrication of a sensitive and quantitative surface-enhanced Raman scattering (SERS)-based lateral flow (LF) strip combined with enzymatic recombinase amplification (ERA) for the highly effective detection of the influenza A virus. Gold-shell silica-core nanoraspberries (NRbs) were used in this SERS-LF strip as a SERS substrate. Practical and convenient detection was accomplished by naked-eye visualization, and highly sensitive quantitative detection was accomplished by monitoring the intensity of the distinct Raman peaks associated with the SERS tags. Under optimal conditions, the sensitivity of this ERA-SERS-LF strip for the DNA standard of the IAV matrix gene is 105 copies/mL by naked-eye visualization, and the limit of detection (LOD) was estimated to be 2.63 × 103 copies/mL by SERS detection, lowered by nearly 40 times compared with visual results due to the signal enhancement of the SERS tags. Besides, quantitative detection for the DNA standard of the IAV matrix gene was obtained with a wide linear range (2.63 × 103 copies/mL to 109 copies/mL). Pseudoviruses of IAV and other respiratory viruses could be successfully identified by using the fabricated ERA-SERS-LF strip. Overall, the findings of this study demonstrate the prospects of the use of the ERA-SERS-LF strip as an effective tool for point-of-care testing (POCT) detection of viruses by virtue of its portability, good sensitivity, quantitative detection ability, and outstanding specificity.
研究领域:
医学诊断
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生物传感
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病毒学
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DOI
高影响因子
Cell Death Discovery
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2025
·Vol. 11
·期 1
·1-13
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IF 7
Wei Cheng,Xu Yitian,Zheng Ying,Hong Lizhe,Lyu Chen,Li Haibo,Cao Bin
The mortality associated with influenza A virus (IAV) infection typically results from excessive immune responses, leading to immunopathological lung damage and compromised pulmonary function. Various immunomodulators are seen beneficial when used in conjunction with direct anti-infection treatment. Leukotriene B4 (LTB4) is a derivative of arachidonic acid (AA) and has been shown to be advantageous for numerous infectious diseases, allergies, and autoimmune disorders. Nonetheless, the function of LTB4 in influenza infection remains unclear. This study demonstrates that LTB4 and its primary receptor BLT1, as opposed to the secondary receptor BLT2, act as a protective immune modulator during influenza infection in bone marrow-derived macrophages and mouse models. Mechanistically, LTB4 promotes K27-linked and K48-linked polyubiquitination of the NLRP3 protein at its K886 and K1023 sites via a cAMP/PKA-dependent pathway, which inhibits NLRP3 inflammasome assembly and thereby diminishes subsequent NLRP3 inflammasome activation. The consequent decline in the release of IL-1β and IL-18 leads to a reduction in inflammation caused by viral infection. Furthermore, the administration of a LTB4 treatment in a fatal IAV infection model can mitigate the excessive NLRP3 inflammasome activation and reduce IAV-induced severe pulmonary damage. These findings illustrate the protective function of LTB4 in fatal IAV infection by mitigating the severe inflammation induced by the virus.
研究领域:
免疫学
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病毒学
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药理学
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DOI
PubMed
高影响因子
Frontiers in Cellular and Infection Microbiology
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2022
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IF 6.073
Cui Huan,Zhang Cheng,Zhang Chunmao,Cai Zhuming,Chen Ligong,Chen Zhaoliang,Zhao Kui,Qiao Sina,Wang Yingchun,Meng Lijia,Dong Shishan,Liu Juxiang,Guo Zhendong
Influenza virus is a serious threat to global human health and public health security. There is an urgent need to develop new anti-influenza drugs. Lentinan (LNT) has attracted increasing attention in recent years. As potential protective agent, LNT has been shown to have anti-tumor, anti-inflammatory, and antiviral properties. However, there has been no further research into the anti-influenza action of lentinan in vivo, and the mechanism is still not fully understood. In this study, the anti-influenza effect and mechanism of Lentinan were studied in the Institute of Cancer Research (ICR) mouse model. The results showed that Lentinan had a high degree of protection in mice against infection with influenza A virus, delayed the emergence of clinical manifestations, improved the survival rate of mice, significantly prolonged the middle survival days, attenuated the weight loss, and reduced the lung coefficient of mice. It alleviated the pathological damage of mice infected with the influenza virus and improved blood indices. Lentinan treatment considerably inhibited inflammatory cytokine (TNF-α, IL-1β, IL-4, IL-5, IL-6) levels in the serum and lung and improved IFN-γ cytokine levels, which reduced cytokine storms caused by influenza virus infection. The underlying mechanisms of action involved Lentinan inhibiting the inflammatory response by regulating the TLR4/MyD88 signaling pathway. This study provides a foundation for the clinical application of Lentinan, and provides new insight into the development of novel immunomodulators.
研究领域:
免疫学
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病毒学
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药理学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
Evidence-based Complementary and Alternative Medicine
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2021
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IF 2.63
Xin Mao,Shuyin Gu,Huiting Sang,Yilu Ye,Jingyan Li,Yunxia Teng,Feiyu Zhang,Qinhai Ma,Ping Jiang,Zifeng Yang,Weizhong Huang,Shuwen Liu
Background. Luofushan-Baicao Oil (LBO) is an essential oil-rich traditional Chinese medicine (TCM) formula that is commonly used to treat cold, cough, headache, sore throat, swelling, and pain. However, the anti-influenza activities of LBO and the underlying mechanism remain to be investigated. Methods. The in vitro anti-influenza activity of LBO was tested with methyl thiazolyl tetrazolium (MTT) and plaque assays. The effects of LBO on the expressions of viral nucleoprotein and cytokines were evaluated. In the polyinosinic-polycytidylic acid- (Poly I: C-) induced inflammation model, the influences of LBO on the expression of cytokines and the activation of NF-κB P65 (P65) and interferon regulatory factor 3 (IRF3) were tested. After influenza A virus (IVA) infection, mice were administered with LBO for 5 days. The lung index, histopathologic change, the expression of viral protein, P65, and IRF3 in the lung tissue were measured. The levels of proinflammatory cytokines in serum were examined. Results. In vitro, LBO could significantly inhibit the infection of IVA, decrease the formation of plaques, and reduce the expression of viral nucleoprotein and cytokines. LBO could also effectively downregulate the expression of interleukin-1β (IL-1β), interleukin-6 (IL-6), and interferon-β and the activation of P65 and IRF3 in Poly I:C-treated cells. In the IVA-infected mice model, inhalation of LBO with atomizer could decrease the lung index, alleviate the pathological injury in the lung tissue, and reduce the serum levels of IL-1β and IL-6. LBO could significantly downregulate the expression of viral protein (nucleoprotein, PB2, and matrix 2 ion channel) and the phosphorylation of P65 and IRF3 in the lungs of mice. Conclusion. The therapeutic effects of LBO on treating influenza might result from the regulation of the immune response of IVA infection. LBO can be developed as an alternative therapeutic agent for influenza prevention.
研究领域:
药理学
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免疫学
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病毒学
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中医药
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DOI
PubMed
高影响因子
Journal of Pharmaceutical Analysis
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2025
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IF 8.9
Yuexiang CHANG,Hao TIAN,Jia-Huan QU,Jiaming YUAN,Rongkai GU,TingTing ZHANG,Jincai WANG,Zhengjin JIANG
The N -terminal domain of influenza viral polymerase (PA N ), a highly conserved region with critical catalytic function related to viral RNA replication and transcription, is considered as a very promising anti-influenza drug target. There is an urgent need for highly efficient and rapid screening methods to identify potential PA N inhibitors (PA N Is) from complex matrices. In this work, a novel high-throughput screening (HTS) platform was established by coupling high performance liquid chromatography and high-resolution mass spectrometry (HPLC-HRMS) with a fluorescence resonance energy transfer (FRET)-based endonuclease activity assay through an at-line nanofractionation system (ANF). The proposed screening platform could rapidly identify potential PA N Is from plant extracts with good sensitivity (baloxavir at the half maximal inhibitory concentration (IC 50 ) could be detected) and reliability ( Z ’ factor of 0.77). This platform was then successfully applied to the screening of potential inhibitors against PA N /PA N I38T from an aqueous extract of Artemisiae Argyi and 17 potential PA N Is were identified. Among them, three compounds (cynarine, isochlorogenic acid B and isochlorogenic acid C) showed comparable inhibitory activity against PA N and even better activity against PA N I38T compared to baloxavir. This study not only established a novel high-throughput ANF-based PA N Is screening platform, but also proved the feasibility to discover PA N Is from complex TCMs, which has a great potential in future anti-influenza drug discovery.
研究领域:
病毒学
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药物筛选
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植物药理学
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生物分析技术
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产品关联
DOI
PubMed
高影响因子
Heliyon
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2024
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IF 3.4
Xin Yang,Junxian Li,Chunlan Shan,Xuqin Song,Jian Yang,Hao Xu,Deyuan Ou
In the present study, we investigated whether baicalin could reduce the damage caused to RAW264.7 cells following infection with H6N6 avian influenza virus. In addition, we studied the expression of autophagy-related genes. The morphological changes in cells were observed by hematoxylin and eosin (H&E) staining, and the inflammatory factors in the cell supernatant were detected by enzyme-linked immunosorbent assay (ELISA). Transmission electron microscopy (TEM) was used to detect the levels of RAW264.7 autophagosomes, and western blotting and immunofluorescence were used to detect the protein expression of autophagy marker LC3. Quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) was used to detect the mRNA transcription levels of autophagy key factors. The results showed that different doses of baicalin significantly reduced the H6N6 virus-induced damage of RAW264.7 cells. The contents of interleukin (IL)-1β, IL-2, IL-6, and tumor necrosis factor (TNF)-α in the cell supernatant significantly decreased. In addition, the protein expression of LC3 and Beclin-1, ATG12, ATG5 the mRNA levels were significantly decreased. This study showed that baicalin can reduce cell damage and affect the H6N6-induced autophagy level of RAW264.7 cells.
研究领域:
病毒学
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药理学
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免疫学
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产品关联
DOI
PubMed
高影响因子
EBioMedicine
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2022
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IF 11.205
Mengyi Zhang,Yimeng An,Xi Wu,Meina Cai,Xinyu Zhang,Chaoying Yang,Jincheng Tong,Zhimin Cui,Xueli Li,Weijin Huang,Chenyan Zhao,Youchun Wang
Summary Background Influenza A (H3N2) virus (A/H3N2) has complex antigenic evolution, resulting in frequent vaccine strain updates. We aimed to evaluate the protective effect of the vaccine strains on the circulating strains from past ten years and provide a basis for finding a broader and more efficient A/H3N2 vaccine strain. Methods Eighty-four representative circulating A/H3N2 strains were selected from 65,791 deposited sequences in 2011–2020 and pseudotyped viruses were constructed with the VSV vector. We immunized guinea pigs with DNA vaccine containing the A/H3N2 components of the vaccine strains from 2011 to 2021 and tested neutralizing antibody against the pseudotyped viruses. We used a hierarchical clustering method to classify the eighty-four representative strains into different antigenic clusters. We also immunized animals with monovalent vaccine stock of the vaccine strains for the 2020–2021 and 2021–2022 seasons and tested neutralizing antibody against the pseudotyped viruses. Findings The vaccine strains PE/09, VI/11 and TE/12 induced higher levels of neutralizing antibody against representative strains circulating in recommended year and the year immediately prior whereas vaccine strains HK/14, HK/19 and CA/20 induced poor neutralization against all representative strains. The representative strains were divided into five antigenic clusters (AgV), which were not identical to gene clades. The AgV5 strains were most difficult to be protected among the five clusters. Compared with single-dose immunization, three doses of monovalent vaccine stock (HK/19 or CA/20) could induce stronger and broader neutralizing antibodies against strains in each of the antigenic clusters. Interpretation The protective effect of vaccine strains indicated that the accurate selection of A/H3N2 vaccine strains must remain a top priority. By increasing the frequency of immunization, stronger and broader neutralizing antibodies against strains in all antigenic clusters were induced, which provides direction for a new immunization strategy. Funding This work was supported by a grant from National Key R&D Program of China (No. 2021YFC2301700 ).
研究领域:
病毒学
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免疫学
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流行病学
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DOI
PubMed
高影响因子
JOURNAL OF VIROLOGICAL METHODS
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2014
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IF 1.883
Yuan-e Yang,Junlin Wen,Suqing Zhao,Kun Zhang,Yingliang Zhou
Influenza A virus infects the human respiratory system and causes acute and fatal pulmonary diseases. The emergence of drug-resistant viral strains highlights the need for alternative therapeutic approaches. In this work, IgY antibody was raised in immunized laying hens, and its antiviral activity was evaluated in the context of passive immunization. With inactivated whole H1N1 virus, high-titer IgY antibody 9.18 mg/mL egg yolk was induced by the eighth week after immunization. Western blotting and the hemagglutination inhibition (HI) test demonstrated that the IgY antibody could specifically bind the neuraminidase and hemagglutinin of the H1N1 virus. In the plaque reduction assay, the IgY antibody reduced the H1N1 viral infection in MDCK (Madin-Darby canine kidney) cells. In a mouse model, the anti-H1N1 IgY antibody exhibited in vivo protection by reducing the infectious titer of the virus in the lung while maintaining the weight and normal structure of the lung tissue. Additionally, the anti-H1N1 IgY antibody exhibited protective activity comparable to the neuraminidase inhibitor oseltamivir. These results demonstrated that IgY can be easily produced and can offers an effective alternative approach for influenza control.
研究领域:
免疫学
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病毒学
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医学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF VIROLOGY
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2021
·Vol. 95
·期 3
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IF 5.103
Ning Ma,Xingjie Li,Hongyu Jiang,Yulong Dai,Guofeng Xu,Zongde Zhang
Influenza A viruses cause severe respiratory illnesses in humans and animals. Overreaction of the innate immune response to influenza virus infection results in hypercytokinemia, which is responsible for mortality and morbidity. However, the mechanism by which influenza induces hypercytokinemia is not fully understood. In this study, we established a mouse-adapted H9N2 virus, MA01, to evaluate the innate immune response to influenza in the lung. MA01 infection caused high levels of cytokine release, enhanced pulmonary injury in mice, and upregulated CD83 protein in dendritic cells and macrophages in the lung. Influenza virus neuraminidase (NA) unmasked CD83 protein and contributed to high cytokine levels. Furthermore, we provide evidence that CD83 is a sialylated glycoprotein. Neuraminidase treatment enhanced lipopolysaccharide (LPS)-stimulated NF-κB activation in RAW264.7 cells. Anti-CD83 treatment alleviated influenza virus-induced lung injury in mice. Our study indicates that influenza virus neuraminidase modulates CD83 status and contributes to the “cytokine storm,” which may suggest a new approach to curb this immune injury.IMPORTANCE The massive release of circulating mediators of inflammation is responsible for lung injury during influenza A virus infection. This phenomenon is referred to as the “cytokine storm.” However, the mechanism by which influenza induces the cytokine storm is not fully understood. In this study, we have shown that neuraminidase unmasked CD83 protein in the lung and contributed to high cytokine levels. Anti-CD83 treatment could diminish immune damage to lung tissue. The NA-CD83 axis may represent a target for an interruption of influenza-induced lung damage.
研究领域:
免疫学
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病毒学
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医学
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DOI
PubMed
高影响因子
MICROCHEMICAL JOURNAL
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2024
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IF 4.9
Yuqi Yang,Shengjun Bu,Xiaoying Zhang,Qiqing Duan,Heyuan Meng,Zhuo Hao,Xiuxia He,Jiayu Wan
Infectious diseases caused by the influenza virus are the primary global concern. This study aimed to develop an electrochemical biosensor for detecting influenza A H1N1 virus based on duplex-specific nuclease (DSN) and rolling circle amplification (RCA) technologies. A single-stranded DNA (ssDNA) probe, comprising a recognition sequence for a target sequence of influenza virus H1N1-specific RNA sequence and primer sequence for initiating RCA, was designed and synthesized. A specific sequence of target H1N1 RNA triggered cyclical DSN cleavage of the probe. The released primer sequence then initiated RCA to generate detectable ssDNA-labeled multiple biotins. ssDNA was captured by streptavidin (SA) on the electrode surface and labeled with free Streptavidin-Horseradish Peroxidase (SA-HRP). The H1N1 influenza virus was quantitatively analyzed based on the changing current signal generated on the electrode surface. A linear range from 10 pM to 100 nM and a limit of detection of 0.44 pM were recorded. In addition, the biosensor demonstrated excellent selectivity, repeatability, and stability for analyzing H1N1 in real samples. This biosensor could be applied for the clinical diagnosis of influenza A H1N1 virus.
研究领域:
医学诊断
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分子生物学
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生物传感技术
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产品关联
DOI
高影响因子
JOURNAL OF ETHNOPHARMACOLOGY
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2025
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IF 5.4
Jiawang Huang,Jianing Shi,Jingmin Fu,Zhuolin Liu,Zhiying Feng,Kangyu Wang,Chunjing Chen,Yi Ning,Fangguo Lu,Ling Li
Ethnopharmacology relevance Maxing Shigan Decoction (MXSGD), derived from the traditional Chinese medicine (TCM) classic Shang Han Lun , is recognized as an effective TCM herbal formula for treating respiratory diseases, although its precise molecular mechanisms against influenza A virus (IAV) have not yet been fully elucidated. Aim of the study This study aimed to clarify the pharmacological effects of MXSGD on IAV-infected pneumonia mouse models, with a particular focus on its regulatory mechanism through miR-1260. Materials and methods The chemical composition of MXSGD was analyzed using an ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). After establishing IAV-infected mouse models, the pathological changes in lung tissues were observed by hematoxylin-eosin (HE) staining; the pulmonary epithelial cells and macrophages in lung tissues were examined through immunological assays; and the expression levels of miR-1260 and its target genes in lung tissues were evaluated by RT-qPCR and Western blot analysis. A co-culture system of MLE-12 and RAW246.7 cells was constructed as the research model. Then, the intracellular miR-1260 expression was detected by RT-qPCR, and the Sema3A expression was examined by immunofluorescence. The macrophages were further transfected with miR-1260 mimic, inhibitor, and Sema3A siRNA to determine whether MXSGD exerts its therapeutic effects through the miR-1260-targeted Sema3A-mediated PI3K/AKT/mTOR signaling pathway. Subsequently, alveolar macrophage-depleted mouse models were established using CL2MBP, and were infected with IAV. To clarify the role of macrophages in the therapeutic mechanism of MXSGD against IAV, the body weight and organ indices of mice were monitored and recorded; the total protein content and levels of inflammatory factors in the bronchoalveolar lavage fluid (BALF) were measured; the histopathological changes were observed by HE staining; the pulmonary epithelial cells and macrophages in lung tissues were examined through immunological assays; and the expressions of miR-1260 and its target genes in lung tissues were evaluated by RT-qPCR and Western blot analysis. Results MXSGD was found to alleviate IAV-induced pathological injury and suppress macrophage recruitment in mouse lung tissues, while downregulating the viral load and levels of inflammatory factors. The attenuation of macrophage recruitment was closely associated with pulmonary epithelial cells, potentially mediated through the miR-1260-targeted Sema3A-regulated PI3K/AKT/mTOR signaling pathway. In cellular experiments, both cell types in the co-culture system were restored following MXSGD treatment, accompanied by lowered intracellular viral load and levels of inflammatory factors. Additionally, macrophage apoptosis and miR-1260 expression were significantly reduced, whereas the Sema3A fluorescence intensity in macrophages was markedly increased. By transfecting miR-1260 mimic, inhibitor, and Sema3A siRNA into co-cultured macrophages, the therapeutic effects of MXSGD were further confirmed to be associated with the miR-1260/Sema3A-mediated PI3K/AKT/mTOR pathway. In IAV-infected models established after pulmonary macrophage depletion, the therapeutic efficacy of MXSGD was partially weakened following elimination of alveolar macrophages. In general, above in vivo findings are consistent with the cellular experimental results, collectively supporting the involvement of the miR-1260-targeted Sema3A-mediated PI3K/AKT/mTOR signaling pathway in the therapeutic mechanism of MXSGD. Conclusions MXSGD was demonstrated to regulate the PI3K/AKT/mTOR pathway through the macrophage miR-1260/Sema3A axis, through which excessive inflammatory responses in IAV-induced pneumonia were effectively suppressed. The findings provide a novel therapeutic target for antiviral treatment in TCM.
研究领域:
免疫学
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病毒学
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分子生物学
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呼吸病学
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中药学
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产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）