Retinoic acid induced specific changes in the phosphoproteome of C17.2 neural stem cells.

Retinoic acid (RA), a vitamin A derivative, is an effective cell differentiating factor which plays critical roles in neuronal differentiation induction and the production of neurotransmitters in neurons. However, the specific changes in phosphorylation levels and downstream signalling pathways associated with RA remain unclear. This study employed qualitative and quantitative phosphoproteomics approaches based on mass spectrometry to investigate the phosphorylation changes induced by RA in C17.2 neural stem cells (NSCs). Dimethyl labelling, in conjunction with TiO2 phosphopeptide enrichment, was utilized to profile the phosphoproteome of self-renewing and RA-induced differentiated cells in C17.2 NSCs. The results of our study revealed that, qualitatively, 230 and 14 phosphoproteins were exclusively identified in the self-renewal and RA-induced groups respectively. Quantitatively, we successfully identified and quantified 177 unique phosphoproteins, among which 70 exhibited differential phosphorylation levels. Analysis of conserved phosphorylation motifs demonstrated enrichment of motifs corresponding to cyclin-dependent kinase and MAPK in the RA-induced group. Additionally, through a comprehensive literature and database survey, we found that the differentially expressed proteins were associated with the Wnt/ß-catenin and Hippo signalling pathways. This work sheds light on the changes in phosphorylation levels induced by RA in C17.2 NSCs, thereby expanding our understanding of the molecular mechanisms underlying RA-induced neuronal differentiation.

Landscape of clinical drug development of ADCs used for the pharmacotherapy of cancers: an overview of clinical trial registry data from 2002 to 2022.

BACKGROUND: To provide reference for clinical development of ADCs in the industry, we analyzed the landscape and characteristics of clinical trials about antibody-drug conjugates (ADCs). METHOD: Clinical trials to study ADCs used for the pharmacotherapy of cancers initiated by the sponsor were searched in the Cite line Pharma Intelligence (Trialtrove database), and the landscape and characteristics of these clinical trials were analyzed from multiple perspectives, such as the number, phases, status, indications, and targets of the clinical trials. RESULT: As of December 31, 2022, a total of 431 clinical trials have been initiated to study ADCs used for the pharmacotherapy of cancers, and the number of the last 10 years was 5.5 times as large as the first 11 years. These clinical trials involved 47 indications, including breast cancer, lymphoma (lymphoma, non-Hodgkin's and lymphoma, Hodgkin's), unspecified solid tumor, bladder cancer and lung cancer (lung, non-small cell cancer and lung, small cell cancer). As for each of these five indications, 50 + clinical trials have been carried out, accounting for as high as 48.50% (454/936). ADCs involve 38 targets, which are relatively concentrated. Among them, ERBB2 (HER2) and TNFRSF8 (CD30) involve in 100 + registered clinical trials, and TNFRSF17 (BCMA), NECTIN4 and CD19 in 10 + trials. The clinical trials for these five targets account for 79.02% (354/448) of the total number. Up to 93.97% (405/431) of these clinical trials explored the correlation between biomarkers and efficacy. Up to 45.91% (292/636) of Lots (lines of treatment) applied in the clinical trials were the second line. Until December 31, 2022, 54.52% (235/431) of the clinical trials have been completed or terminated. CONCLUSION: ADCs are a hotspot of research and development in oncology clinical trials, but the indications, targets, phases, and Lot that have been registered are seemingly relatively concentrated at present. This study provides a comprehensive analysis which can assist researchers/developer quickly grasp relevant knowledge to assess a product and also providing new clues and ideas for future research.

A phosphoproteomics study reveals a defined genetic program for neural lineage commitment of neural stem cells induced by olfactory ensheathing cell-conditioned medium.

Since both Olfactory ensheathing cells (OECs) and neural stem cells (NSCs) have shown certain efficacy in the cellular therapy of nerve injury and disease, there have been a series of investigations in recent years looking at the co-culture of NSCs and OECs. Protein phosphorylation forms the basis for identifying a variety of cellular signaling pathways responsible for regulating the self-renewal and differentiation of NSCs induced by OECs. To better understand the signaling cascades in the early phases of OEC-induced NSC differentiation, changes in the NSC proteome and phosphoproteome during the first 24 h were determined using dimethyl labeling and TiO2 phosphorylation enrichment coupled with Liquid chromatography-tandem mass spectrometry (LC-MS/MS). A total of 565 proteins and 2511 phosphorylation sites were identified. According to quantitative phosphoproteomics analyses of NSC differentiation induced by OECs during the first 12 and 24 h, it was speculated that there were at least two different signal waves: one peaking within 12 h after stimulation and the second upsurge after 24 h. In addition to understanding the dynamics of the proteome and phosphoproteome in the early stages of NSC differentiation, our analyses identified a key role of the TGF-ß3 protein secreted by OECs, which may be an initiating factor that promotes differentiation of NSCs into neurons induced by OECs. These findings not only redemonstrated a OECs-based therapeutic strategy in cell therapy, but also added a node to the regulatory network for the neural lineage commitment of NSCs induced by OECs.

Worldwide aflatoxin contamination of agricultural products and foods: From occurrence to control.

Aflatoxins represent a global public health and economic concern as they are responsible for significant adverse health and economic issues affecting consumers and farmers worldwide. Produced by fungal species from the Aspergillus genus, aflatoxins are a toxic, mutagenic, and carcinogenic group of fungal metabolites that routinely contaminate food and agricultural products. Climate and diet are essential factors in the aflatoxin contamination of food and subsequent human exposure process. Countri es with warmer climates and staple foods that are aflatoxin-susceptible shoulder a substantial portion of the global aflatoxins burden. Enactment of regulations, prevention of pre- and postharvest contamination, decontamination, and detoxification have been used to prevent human dietary exposure to aflatoxin. Exploiting their chemical and structural properties, means are devised to detect and quantify aflatoxin presence in foods. Herein, recent developments in several important aspects impacting aflatoxin contamination of the food supply, including: fungal producers of the toxin, occurrence in food, worldwide regulations, detection methods, preventive strategies, and removal and degradation methods were reviewed and presented. In conclusion, aflatoxin continues to be a major food safety problem, especially in developing countries where regulatory limits do not exist or are not adequately enforced. Finally, knowledge gaps and current challenges in each discussed aspect were identified, and new solutions were proposed.

Quantitative phosphoproteomics of lectin receptor-like kinase VI.4 dependent abscisic acid response in Arabidopsis thaliana.

Lectin receptor-like kinases (LecRKs) play important roles in the responses to adverse environment stress. Abscisic acid (ABA) is a plant hormone involved in plant growth, development and adverse environmental stress responses. Although some studies of ABA response LecRK genes have been reported, the molecular mechanisms of LecRKs regulation of downstream pathways under ABA induction are not well understood. The present study showed that LecRK-VI.4 responded to ABA and negatively regulated stomatal closure. Here, a quantitative phosphoproteomics approach based on mass spectrometry was employed to study the roles of LecRK-VI.4 in the ABA signaling pathway. Metal oxide affinity beads and C18 chromatography were used for phosphopeptide enrichment and separation. The isobaric tags for relative and absolute quantitation were used for profiling the phosphoproteome of mutant lecrk-vi.4-1 and wild-type Col-0 Arabidopsis under normal growth conditions or ABA treatments. In total, 475 unique phosphopeptides were quantified, including 81 phosphopeptides related to LecRK-VI.4 regulation. Gene ontology, protein-protein interaction and motif analysis were performed. The bioinformatics data showed that phosphorylated proteins regulated by LecRK-VI.4 had close relations with factors of stomatal function, which included aquaporin activity, H+ pump activity and the Ca2+ concentration in the cytoplasm. These data have expanded our understanding of how LecRK-VI.4 regulates ABA-mediated stomatal movements.

Specific antigen-based and emerging detection technologies of mycotoxins.

Mycotoxins are secondary fungal metabolites produced by certain types of filamentous fungi or molds, such as Aspergillus, Fusarium, Penicillium, and Alternaria spp. Mycotoxins are natural contaminants of agricultural commodities, and their prevalence may increase due to global warming. According to the Food and Agriculture Organization of the United Nations, approximately 25% of the world's food crops are annually contaminated with mycotoxins. Mycotoxin-contaminated food and feed pose a high risk to both human and animal health. For instance, they possess carcinogenic, immunosuppressive, hepatotoxic, nephrotoxic, and neurotoxic effects. Hence, various approaches have been used to assess and control mycotoxin contamination. Significant challenges still exist because of the complex heterogeneous nature of food and feed composition. The potential of antigen-based approaches, such as enzyme-linked immunosorbent assay, flow injection immunoassay, chemiluminescence immunoassay, lateral flow immunoassay, and flow-through immunoassay, would contribute to our understanding about mycotoxins' rapid identification, their isolation, and the basic principles of the detection technologies. Additionally, we address other emerging technologies of potential application in the detection of mycotoxins. The data included in this review focus on basic principles and results of the detection technologies and would be useful as benchmark information for future research. © 2019 Society of Chemical Industry.

Monitoring Metabolite Production of Aflatoxin Biosynthesis by Orbitrap Fusion Mass Spectrometry and a D-Optimal Mixture Design Method.

Aflatoxins, highly toxic and carcinogenic to humans, are synthesized via multiple intermediates by a complex pathway in several Aspergilli, including Aspergillus flavus. Few analytical methods are available for monitoring the changes in metabolite profiles of the aflatoxin biosynthesis pathway under different growth and environmental conditions. In the present study, we developed by a D-optimal mixture design a solvent system, methanol/dichloromethane/ethyl acetate/formic acid (0.36/0.31/0.32/0.01), that was suitable for extracting the pathway metabolites. The matrix effect from dilution of cell extracts was negligible. To facilitate the identification of these metabolites, we constructed a fragmentation ion library. We further employed liquid chromatography coupled with high-resolution mass spectroscopy (UHPLC-HRMS) for simultaneous quantification of the metabolites. The limit of detection (LOD) and limit of quantitation (LOQ) were 0.002-0.016 and 0.008-0.05 µg/kg, respectively. The spiked recovery rates ranged from 81.3 to 100.3% with intraday and interday precision less than 7.6%. Using the method developed to investigate the time-course aflatoxin biosynthesis, we found that precursors, including several possible toxins (with a carcinogenic group similar to aflatoxin B1), occurred together with aflatoxin, and that production increased rapidly at the early growth stage, peaked on day four, and then decreased substantially. The maximum production of aflatoxin B1 and aflatoxin B2 occurred 1 day later. Moreover, the dominant branch pathway was the one for aflatoxin B1 formation. We revealed that the antiaflatoxigenicity mechanism of Leclercia adecarboxylata WT16 was associated with a factor upstream of the aflatoxin biosynthesis pathway. The design strategies can be applied to characterize or detect other secondary metabolites to provide a snapshot of the dynamic changes during their biosynthesis.

Multispecies Adulteration Detection of Camellia Oil by Chemical Markers.

Adulteration of edible oils has attracted attention from more researchers and consumers in recent years. Complex multispecies adulteration is a commonly used strategy to mask the traditional adulteration detection methods. Most of the researchers were only concerned about single targeted adulterants, however, it was difficult to identify complex multispecies adulteration or untargeted adulterants. To detect adulteration of edible oil, identification of characteristic markers of adulterants was proposed to be an effective method, which could provide a solution for multispecies adulteration detection. In this study, a simple method of multispecies adulteration detection for camellia oil (adulterated with soybean oil, peanut oil, rapeseed oil) was developed by quantifying chemical markers including four isoflavones, trans-resveratrol and sinapic acid, which used liquid chromatography tandem mass spectrometry (LC-MS/MS) combined with solid phase extraction (SPE). In commercial camellia oil, only two of them were detected of daidzin with the average content of 0.06 ng/g while other markers were absent. The developed method was highly sensitive as the limits of detection (LODs) ranged from 0.02 ng/mL to 0.16 ng/mL and the mean recoveries ranged from 79.7% to 113.5%, indicating that this method was reliable to detect potential characteristic markers in edible oils. Six target compounds for pure camellia oils, soybean oils, peanut oils and rapeseed oils had been analyzed to get the results. The validation results indicated that this simple and rapid method was successfully employed to determine multispecies adulteration of camellia oil adulterated with soybean, peanut and rapeseed oils.

Degradable polyvinyl alcohol/chitosan@carnauba wax superhydrophobic composite membrane for water-in-oil emulsion separation and heavy metal adsorption.

At present, many oil-water separation membranes are being developed to purify oily wastewater. However, oily wastewater often contains heavy metal, which are often difficult to dispose during separation. Furthermore, most of the oil-water separation membranes cannot be degraded after scrap, producing pollution to environment. Herein, the polyvinyl alcohol/chitosan@carnauba wax (PCGCW) membrane with heavy metal adsorption and biodegradation performance was acquired by electrospinning and spraying process. The acquired PCGCW membrane had excellent mechanical properties after crosslinking glutaraldehyde (GA). Furthermore, the composite membrane had excellent superhydrophobic property (WCA = 154°) with a rolling angle of 2°, due to the introduction of carnauba wax. Exhilaratingly, for emulsions with surfactant, it had a high separation flux with 19,217 L·m-2·h-1·bar-1 and splendid an oil purity over 99.9 %. Besides, the efficiency of oil purity and separation flux remained stable even after 10 separations. In addition, the PCGCW membrane had the ability to adsorb heavy metals with adsorption capacity of 51-106 mg/g for Cu2+, Fe3+, Co2+ ions. Foremost, the superhydrophobic PCGCW membrane was biodegradable, with degrading 29.76 % within 40 days. The prepared composite membrane had the advantages of low cost, high separation flux, great repeatability, adsorbable heavy metals and degradability, which had a vast application prospect.

Protonated cross-linkable nanocomposite coatings with outstanding underwater superoleophobic and anti-viscous oil-fouling properties for crude oil/water separation.

Superhydrophilic/underwater superoleophobic coatings that effectively prevent viscous oil contamination have been of considerable interest for the great potential in oil spill remediation and oilfield wastewater treatment. In the present work, a protonated cross-linkable nanocomposite coating with robust underwater superoleophobicity and intensified hydration capability is proposed through the synthesis of active polymeric nanocomplex (PNC), cross-linking reaction between PNC and hydrophilic chitosan (CS), and final protonation to further improve water affinity. Benefiting from the hierarchical structure and strong hydration capability induced by electrostatic interactions and hydrogen bondings, the nanocomposite coating coated textile exhibits excellent superhydrophilicity (within 0.28 s with water contact angle reaching 0°), underwater superoleophobicity (underwater crude oil contact angle at 160°), and ultralow oil adhesion even to highly viscous silicone oil. Moreover, the nanocomposite coating presents a robust chemical resistance, mechanical tolerance, and storage stability. Simultaneously, the nanocomposite coating adapts well to various porous substrates (e.g., stainless steel mesh and Ni sponge) with great anti-oil-fouling and self-cleaning performances. Importantly, the coating coated textile is successfully applied in crude oil/water separation with excellent efficiency and repeatability. The findings conceivably stand out as a new methodology to fabricate superhydrophilic/underwater superoleophobic materials with outstanding anti-viscous oil-fouling property for practically treating oily wastewater.

Fungi population metabolomics and molecular network study reveal novel biomarkers for early detection of aflatoxigenic Aspergillus species.

Mycotoxins threaten global food safety, public health and cause huge socioeconomic losses. Early detection is an effective preventive strategy, yet efficient biomarkers for early detection of aflatoxigenic Aspergillus species are lacking. Here, we proposed to use untargeted metabolomics and machine learning to mine biomarkers of aflatoxigenic Aspergillus species. We systematically delineated metabolic differences across 568 extensive field sampling A. flavus and performed biomarker analysis. Versicolorin B, 11-hydroxy-O-methylsterigmatocystin et.al metabolites shown a high correlation (from 0.71 to 0.95) with strains aflatoxin-producing capacity. Molecular networking analysis deciphered the connection of aflatoxins and biomarkers as well as potential emerging mycotoxins. We then developed a model using the biomarkers as variables to discern aflatoxigenic Aspergillus species with 97.8% accuracy. A validation dataset and metabolome from other 16 fungal isolates confirmed the robustness and specificity of these biomarkers. We further demonstrated the solution feasibility in agricultural products by early detection of biomarkers, which predicted aflatoxin contamination risk 35-47 days in advance. A developed operable decision rule by the XGBoost algorithm help regulators to intuitively assess the risk prioritization with 87.2% accuracy. Our research provides novel insights into global food safety risk assessment which will be crucial for early prevention and control of mycotoxins.

Skin-inspired thermoelectric nanocoating for temperature sensing and fire safety.

Temperature sensing enables flammable materials to respond intelligently at high temperature, which is conducive to further improving their fire safety. However, it is still challenging to develop a smart nanocoating with sensitive temperature-sensing and efficient flame retardancy. Inspired by human skin, a thermoelectric flame retardant (TE-FR) nanocoating was fabricated by combining a dermis-mimicking thermoelectric (TE) layer and an epidermis-mimicking flame retardant (FR) layer. The TE-FR nanocoating exhibited accurate temperature sensing at 100-300 ℃ and repeatable fire-warning capability. When being burned, the fire-warning response time of the TE-FR nanocoating was only 2.0 s, and it retriggered the fire-warning device within 2.8 s when it was reburned. Meanwhile, the TE-FR nanocoating exhibited outstanding flame retardancy. The coated polypropylene self-extinguished in the horizontal and vertical burning tests. Besides, its peak heat release rate, total heat release, and peak smoke production rate were significantly reduced. This work proposed an ingenious strategy to fabricate smart nanocoating for temperature sensing and fire safety, which revealed an enticing prospect in the fields of fire protection, electronic skin, and temperature monitor.

Aspergillus flavus's Response to Antagonism Bacterial Stress Sheds Light on a Regulation and Metabolic Trade-Off Mechanism for Adversity Survival.

Biocontrol to combat the menace of Aspergillus flavus has gained considerable attention. However, the molecular mechanisms of A. flavus 's response to antagonism biotic stress are poorly deciphered. Here, we discovered that A. flavus switches an adaptive metabolic reprogramming to ensure its adversity survival by multiomics analyses (including four omics platform). Antifungal "weapons" lipopeptides and antibacterial metabolites of imizoquin were identified. The central metabolism fluxes were significantly depleted but the expressions of most corresponding genes were considerably increased in A. flavus. Secondary metabolism that does not contribute to stress was markedly suppressed. In contrast, A. flavus antibacterial "weapon arsenal" was activated to occupy an ecological niche. Our results revealed that interlinked mitochondrial central metabolism and secondary metabolism are central to A. flavus antagonism biotic stress response. This discovery contributes to the targeted design of biocontrol agents and smart regularization of rhizosphere microbiome homeostasis to realize long-term fungi pathogen control and mitigation mycotoxin contamination.

A green approach to fabricating nacre-inspired nanocoating for super-efficiently fire-safe polymers via one-step self-assembly.

Developing a high efficient, environmental-friendly and universal fire-safe strategy for combustible polymers is crucial but challengeable. Inspired by nacre, we developed a super-efficiently fire-safe nanocoating based on carboxymethyl chitosan (CCS) and modified montmorillonite (MMT) via one-step self-assembly. The nanocoating possessed well-arranged nacre-like hierarchical microstructure, exhibiting high transparency and specific nacre-like iridescence. More importantly, the nanocoating endowed many large-scale polymer substrates, such as polyester film, cotton fabric and polyurethane foam, with super-efficient fire-safety by dip-coating or spray-coating. All the coated substrates were self-extinguished in the burning tests. Meanwhile, their heat release and smoke production were decreased remarkably. Most notably, the peak heat release rate, total heat release, peak smoke production rate and total smoke production of polyurethane foam were decreased by 84.1%, 89.4%, 84.4% and 95.2%, respectively. Additionally, no organic solvent, halogen and phosphorus element were involved, which was environmental-friendly. Our findings provide a super-efficient, economical, universal and green fabrication strategy for fire-safe polymers.

Investigation and Characterization of Myroides odoratimimus Strain 3J2MO Aflatoxin B1 Degradation.

Aflatoxin B1 (AFB1), is a type I carcinogen that is one of the strongest naturally occurring aflatoxins and can be injurious to humans and livestock upon ingestion, inhalation, or skin contact, with carcinogenic and mutagenic effects. It causes significant hazardous effects to the food- and animal-production industries. We found a bacterial strain, 3J2MO, that degraded AFB1 well, and here we tested and characterized its AFB1-degradation ability. The strain degraded about 93.82% of the AFB1 after incubation for 48 h in Luria-Bertani (LB) medium at 37 °C with a final concentration of 100 ppb and an inoculation quantity of 1 × 107 cfu/mL. High-performance liquid chromatography-fluorescence detection (HPLC-FLD) was used to determine AFB1 amounts. The maximum degradation rates were 89.23% at pH 8.5; 55.78% at an inoculation quantity of 1 × 108 cfu/mL; and 71.50 and 71.21% at 34 and 37 °C, respectively. Treatment with sucrose and soluble starch as carbon sources and beef extract and ammonium acetate as nitrogen sources stimulated the degradation rate. Mg2+ and Ca2+ ions were activators for AFB1 degradation; however, Mn2+, Fe3+, Zn2+, and Cu2+ were strong inhibitors. This bacterial strain has potential in bioremediation and the detoxification of aflatoxin contamination for biocontrol strategies in both agricultural products and food-industry matrices.

Mechanistic insights into Nav1.7-dependent regulation of rat prostate cancer cell invasiveness revealed by toxin probes and proteomic analysis.

Voltage-gated sodium channels are involved in tumor metastasis, as potentiating or attenuating their activities affects the migration and invasion process of tumor cells. In the present study, we tested the effect of two peptide toxins, JZTX-I and HNTX-III which function as Nav1.7 activator and inhibitor, respectively, on the migration and invasion ability of prostate cancer (PCa) cell line Mat-LyLu. These two peptides showed opposite effects, and subsequently a comparative proteomic analysis characterized 64 differentially expressed membrane proteins from the JZTX-I- and HNTX-III-treated groups. Among these, 15 proteins were down-regulated and 49 proteins were up-regulated in the HNTX-III group. Bioinformatic analysis showed eight proteins are cytoskeleton proteins or related regulators, which might play important roles in the metastasis of Mat-LyLu cells. The altered expressions of four of these proteins, fascin, muskelin, annexin A2, and cofilin-1, were validated by western blot analysis. Further function network analysis of these proteins revealed that the Rho family GTPases RhoA and Rac1 might be of particular importance for the rat PCa cell invasion. Pharmacological data revealed that JZTX-I and HNTX-III could modulate the Rho signaling pathway in a Nav1.7-dependent manner. In summary, this study suggests that the Nav1.7-dependent regulation of Rho GTPase activity plays a vital role in Mat-LyLu cell migration and invasion and provides new insights into the treatment of PCa.

Hypoxic Culture Promotes Dopaminergic-Neuronal Differentiation of Nasal Olfactory Mucosa Mesenchymal Stem Cells via Upregulation of Hypoxia-Inducible Factor-1α.

Olfactory mucosa mesenchymal stem cells (OM-MSCs) display significant clonogenic activity and may be easily propagated for Parkinson's disease therapies. Methods of inducing OM-MSCs to differentiate into dopaminergic (DAergic) neurons using olfactory ensheathing cells (OECs) are thus an attractive topic of research. We designed a hypoxic induction protocol to generate DAergic neurons from OM-MSCs using a physiological oxygen (O2) level of 3% and OEC-conditioned medium (OCM; HI group). The normal induction (NI) group was cultured in O2 at ambient air level (21%). The role of hypoxia-inducible factor-1α (HIF-1α) in the differentiation of OM-MSCs under hypoxia was investigated by treating cells with an HIF-1α inhibitor before induction (HIR group). The proportions of ß-tubulin- and tyrosine hydroxylase (TH)-positive cells were significantly increased in the HI group compared with the NI and HIR groups, as shown by immunocytochemistry and Western blotting. Furthermore, the level of dopamine was significantly increased in the HI group. A slow outward potassium current was recorded in differentiated cells after 21 d of induction using whole-cell voltage-clamp tests. A hypoxic environment thus promotes OM-MSCs to differentiate into DAergic neurons by increasing the expression of HIF-1α and by activating downstream target gene TH. This study indicated that OCM under hypoxic conditions could significantly upregulate key transcriptional factors involved in the development of DAergic neurons from OM-MSCs, mediated by HIF-1α. Hypoxia promotes DAergic neuronal differentiation of OM-MSCs, and HIF-1α may play an important role in hypoxia-inducible pathways during DAergic lineage specification and differentiation in vitro.

Intracellular expression of an anti-idiotypic antibody single-chain variable fragment reduces porcine reproductive and respiratory syndrome virus infection in MARC-145 cells.

BACKGROUND: Porcine reproductive and respiratory syndrome virus (PRRSV) is the causative agent of porcine reproductive and respiratory syndrome; it is one of the most economically important viral diseases affecting the swine industry worldwide. At present, neither live-attenuated nor inactivated PRRSV vaccines can provide sustainable disease control. Our previous studies have demonstrated that PRRSV infection can produce the auto-anti-idiotypic antibodies (aAb2s) specific to the idiotypic antibodies against PRRSV GP5, which plays an important role in the host immune responses to PRRSV infection. In the present study, a single-chain variable antibody fragment (scFv) from the monoclonal anti-idiotypic antibody specific for the idiotypic antibody against GP5 was expressed in MARC-145 cells and its effect on virus infection in vitro was evaluated. METHODS: An scFv was constructed from the anti-idiotypic antibody (Mab2-5G2) and was named 5G2scFv. The lentiviral vector system was used as a vehicle to deliver 5G2scFv into MARC-145 cells. The effect of 5G2scFv expression in MARC-145 was analysed by determining the PRRSV N protein level and the virus titre in the supernatant. Virus attachment and the level of type I interferon (IFN) were determined to elucidate the mechanism of the scFv effect. RESULTS: 5G2scFv was delivered in MARC-145 cells using the lentiviral vector system as confirmed by the western blot and indirect immunofluorescence assays. The PRRSV challenge experiments demonstrated that expressed 5G2scFv in MARC-145 strongly reduced PRRSV infection and replication by inhibiting protein synthesis and progeny virus production. This effect was not due to the change of viability or virus binding, but increased IFN-α at messenger RNA and protein levels. CONCLUSIONS: The expression of the anti-idiotypic antibody 5G2scFv in MARC-145 cells has the interferential effect on PRRSV infection in the cells by induction of IFN-α, which provides a novel therapeutic approach for PRRSV infection.