Mycobacterium tuberculosis infection depressed cytotoxic T cells activity owing to decreasing NKG2C and increasing NKG2A expression.

Cytotoxic T lymphocytes (CTLs) play protective roles in immunity against tuberculosis (TB) infection by strongly inhibiting intracellular mycobacterial growth. In TB infection, the impairing mechanism of CTLs function remains unclear. In this study, we identified that the cytotoxic granule molecules expression levels of perforin (PRF) and granulysin (GNLY) in CD3+ and CD8+ CTL cells were significantly depressed in TB patients compared to those in healthy donors. The frequencies of T-CTLs, co-expressing granzyme B (GZMB), PRF and GNLY, were obviously decreased in TB patients. Moreover, NKG2C highly expressed in T-CTLs, was an effective activator of cytotoxic activity of CD3+ T cells. And, NKG2C+CD3+ T cells potently inhibited intracellular mycobacterial growth. The proportions of NKG2C+ cells in CD3+ and CD8+ T cells were dramatically decreased in TB patients. Contrarily, NKG2A, an inhibitor of T cells cytotoxic activities, was highly expressed in T-CTLs of CD3+ and CD8+ T cells in TB patients. Here, we successfully discovered that depressed CTLs activities in TB patients were attributed to low expression of cytotoxic granule molecules and high expression of inhibitory NKG2A receptor, suppression of agonist receptor NKG2C. Thus, NKG2 receptors were potential targets for immunotherapy of tuberculosis, especially for multidrug-resistant tuberculosis.

Circular RNA circTRAPPC6B Enhances IL-6 and IL-1ß Expression and Repolarizes Mycobacteria Induced Macrophages from M2- to M1-Like Phenotype by Targeting miR-892c-3p.

Mycobacterium tuberculosis (Mtb) infection elicits macrophage polarization into M2 phenotype to block the host's protective immune response. However, it remains unclear how Mtb regulates macrophage polarization. Recent studies have suggested that noncoding RNA may play a role in macrophage polarization. In this study, we investigated the potential involvement of circTRAPPC6B, a circular RNA that is downregulated in tuberculosis (TB) patients, in regulating macrophage polarization. We found that Mtb infection downregulated M1-related IL-6 and IL-1ß while highly expressed M2-related CCL22 and CD163. Overexpressed circTRAPPC6B had switched Mtb-infected macrophages from M2- to M1-like phenotype, accompanied by upregulation of IL-6 and IL-1ß. Meanwhile overexpressed circTRAPPC6B significantly inhibited Mtb growth in macrophages. Our findings suggest that circTRAPPC6B may regulate macrophage polarization by targeting miR-892c-3p, which is highly expressed in TB patients and M2-like macrophages. And miR-892c-3p inhibitor decreased intracellular Mtb growth in macrophages. Thus, TB-inhibited circTRAPPC6B could specifically induce IL-6 and IL-1ß expression to switch/antagonize Mtb-induced macrophage polarization from M2- to M1-like phenotype by targeting miR-892c-3p, leading to enhanced host clearance of Mtb. Our results reveal a potential role for circTRAPPC6B in regulating macrophage polarization during Mtb infection and provide new insights into the molecular mechanisms underlying host defense against Mtb.

Hormesis effects of sulfoxaflor on Aphis gossypii feeding, growth, reproduction behaviour and the related mechanisms.

Sulfoxaflor, an important alternative insecticide in integrated pest management (IPM) strategies, can effectively control sap-feeding insect pests such as Aphis gossypii. Although the side effects of sulfoxaflor have recently attracted widespread attention, its toxicological characteristics and mechanisms are still largely undefined. Therefore, the biological characteristics, life table and feeding behaviour of A. gossypii were studied to evaluate the hormesis effect of sulfoxaflor. Then, the potential mechanisms of induced fecundity associated with the vitellogenin (Ag. Vg) and vitellogenin receptor (Ag. VgR) genes were investigated. Although the LC10 and LC30 concentrations of sulfoxaflor significantly reduced the fecundity and net reproduction rate (R0) of the directly exposed sulfoxaflor-resistant and susceptible aphids, hormesis effects on fecundity and R0 were observed in the F1 generation of Sus A. gossypii when the parental generation was exposed to the LC10 of sulfoxaflor. Moreover, the hormesis effects of sulfoxaflor on phloem feeding were observed in both A. gossypii strains. Additionally, enhanced expression levels and protein content of Ag. Vg and Ag. VgR were observed in progeny generations when F0 was subjected to the trans- and multigenerational sublethal sulfoxaflor exposure. Therefore, sulfoxaflor-induced resurgence might occur in A. gossypii after exposure to sublethal concentrations. Our study could contribute to a comprehensive risk assessment and provide convincing reference to optimize sulfoxaflor in IPM strategies.

Sulfoxaflor adversely influences the biological characteristics of Coccinella septempunctata by suppressing vitellogenin expression and predation activity.

Sulfoxaflor is a widely used sulfoximine insecticide that has been regarded as an important alternative insecticide for IPM strategies, but a comprehensive study of its potential ecological toxicity is still lacking. In the present work, the growth, longevity, predation and reproduction toxicity of Coccinella septempunctata caused by sulfoxaflor were evaluated. In addition, the potential mechanisms of decreased fecundity in C. septempunctata were investigated by analyzing the transcriptional and protein levels of reproduction-related gene vitellogenin (Vg). In a 20-day acute contact toxicity test, decreased survival proportion, pupation rate, adult emergence ratio, and increased hazard quotient (HQ) values were observed. Moreover, sublethal dosages of sulfoxaflor significantly inhibited the predation, longevity, fecundity and net reproduction rate of progeny. In addition, LR30 of sulfoxaflor dramatically down-regulate the mRNA-expression (F0: 65.38-fold, F1: 2.24-fold) and protein content (F0: 1.35-fold, F1: 1.36-fold) of Vg in the F0 and F1 generations. These results suggested that sulfoxaflor could inhibit the gene and protein content of Vg, thereby reducing the fecundity of C. septempunctata. Our study indicated that sulfoxaflor has potential risks to parent and progeny generations of C. septempunctata. These results provide valuable reference for optimal usage of sulfoxaflor in IPM systems.

Seed treatment with chlorantraniliprole and carbaryl mixture for managing fall armyworm on maize: systemic synergism, control efficiency and synergistic mechanism.

BACKGROUND: Fall armyworm (Spodoptera frugiperda) is one of the major invasive pests in China, and has been widely controlled by labor-intensive foliar sprays of agrochemicals in maize (Zea mays L.). RESULTS: Systemic bioassay showed that mixtures of chlorantraniliprole (Chlor) and carbaryl (Carb) had dramatically synergistic effect on toxicity to S. frugiperda. Particularly, a mixture of Chlor with Carb at a mass ratio of 2:1 (MCC) exhibited the highest toxicity to S. frugiperda. Therefore, seed treatment of Chlor mixed with Carb was studied as a simple, accurate, efficient and low-cost control technology. Our results showed that MCC treatment shortened the median lethal time and 90% lethal time to S. frugiperda compared to Chlor- and Carb-alone treatments. Meanwhile, smaller leaf consumption by S. frugiperda was recorded under MCC treatment compared to Chlor- and Carb-alone treatments. In field trial, maize-seed treatment with MCC showed efficacy up to 39 days post-emergence in preventing S. frugiperda foliar damage at a low infestation pressure. Moreover, chemical quantification by ultrahigh-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) showed that Chlor residues were more absorbed and concentrated in maize leaves of MCC treatment, compared to that of Chlor-alone treatment. CONCLUSION: These results suggested that seed treatment with MCC can be applied to increase the control efficacy and reduce the cost of Chlor-alone treatment for controlling S. frugiperda. The present study provided evidence of an enhanced translocation and accumulation of Chlor residues in maize leaves under MCC treatment, which likely contributed to a synergistic effect against S. frugiperda. © 2022 Society of Chemical Industry.

Molecular characterization of Listeria monocytogenes strains isolated from imported food in China from 14 countries/regions, 2003-2018.

Listeria monocytogenes (Lm) is associated with severe foodborne infections and ubiquitous in the nature. Identification of characteristics of Lm transmission through trading of food products is essential for rapidly tracking Lm sources and controlling dissemination of listeriosis. In this study, a total of 44 Lm strains were isolated from food products originating from 14 countries/regions during 2003-2018 at the Shanghai port. The genomes of these Lm strains were sequenced by high-throughput sequencing. Multilocus sequence typing (MLST) analysis showed that 43 isolates were divided into 17 sequence types (STs). The distribution of STs was decentralized, with the dominant ST2 accounting for only 18.18% of the strains. The LM63 strain did not match with any of the existing STs. Core-genome MLST (cgMLST) analysis based on 1748 core genes categorized the 44 strains into 30 cgMLST types (CTs), with CT10153 and CT7892 as the most predominant CTs. Notably, LM63 and LM67 shared the same CT in the cgMLST analysis. The phylogenetic analysis based on single-copy homologous genes revealed that the 44 Lm strains were primarily classified into two lineages. The SNP analysis also indicated that these strains were roughly divided into two clades, with strains in the first clade mainly collected earlier than those in the second clade, which were predominantly collected from 2010 onwards. The analysis using the virulence factor database (VFDB) indicated that the virulence gene inlJ was the most prevalent among these 44 strains. Notably, ddrA, msbA, and sugC were enriched in this dataset, requiring further clarification of their roles in Listeria through future studies. These results might provide a clue for understanding of the global epidemiology and surveillance of Lm and present insights for implementing effective measures to reduce or prevent Listeria contamination outbreaks in imported food products.

An insecticide phenotypic resistance diagnostic kit for cotton aphid Aphis gossypii Glover (Hemiptera Aphididae).

BACKGROUND: Aphis gossypii is a notorious pest worldwide, and evidence of resistance of A. gossypii to various insecticides has been documented. Diagnostic tools for the rapid and accurate assessment of insecticide resistance are urgently needed to implement effective pest control and insecticide resistance management strategies. RESULTS: Using this diagnostic kit based on the glass vial bioassay, detection results can be obtained in 3 h and the values of 897.86, 133.57, 12 037.45, 2849.26, 19 457.33 and 215.60 ng/cm2 were finally identified as the actual diagnostic doses of imidacloprid, acetamiprid, thiamethoxam, nitenpyram, dinotefuran and sulfoxaflor, respectively. The regression equation between mortalities under diagnostic doses and actual resistance ratios tested by the leaf-dipping method were conducted in different strains of A. gossypii, and the diagnostic mortality of A. gossypii was negatively correlated with the resistance ratio to imidacloprid (r = -0.986, P = 0.002), acetamiprid (r = -0.964, P = 0.008), thiamethoxam (r = -0.930, P = 0.022), nitenpyram (r = -0.950, P = 0.013), dinotefuran (r = -0.976, P = 0.004) and sulfoxaflor (r = -0.937, P = 0.019). Moreover, four A. gossypii field populations were selected to apply the diagnostic kit in the field. CONCLUSIONS: A diagnostic kit based on the glass vial bioassay for the rapid detection of resistance to imidacloprid, acetamiprid, thiamethoxam, nitenpyram, dinotefuran and sulfoxaflor in A. gossypii was developed. The insecticide diagnostic kit for A. gossypii can be a useful screening tool to determine effective insecticides quickly and accurately. © 2022 Society of Chemical Industry.

Comparative Toxicity and Joint Effects of Chlorantraniliprole and Carbaryl Against the Invasive Spodioptera frugiperda (Lepidoptera: Noctuidae).

Fall armyworm, Spodoptera frugiperda, is one of the most devastating invasive pests in China. Chlorantraniliprole (CH) is currently the main agent for controlling S. frugiperda. Carbaryl (CA) has been widely used as a foliar treatment to control S. frugiperda, although the pest has become highly resistant to it. This study investigates the comparative toxicity and joint effects of CH and CA on S. frugiperda. Time-toxicity results showed that CH had high toxicity to 1st and 3rd instar larvae, whereas CA had very low toxicity to 1st and 3rd instar larvae. The mixtures of CH and CA at different mass ratios showed strong synergistic effects on toxicity, and the mass ratio of 2:1 exhibited the highest toxicity to S. frugiperda. Furthermore, the synergistic toxicity of CH and CA at the 2:1 mass ratio (CH+CA) was also verified in field populations of S. frugiperda. The life-history parameters showed that CH+CA dramatically decreased the survival rate and fecundity of the parent population (F0) compared with CH treatment at the same concentration. Besides, CH and CH+CA mixture showed induction effect on cytochrome P450s and glutathione-S-transferases (GSTs) activities in S. frugiperda, with cytochrome P450s enzyme responding the fastest. In conclusion, this research found CH+CA provided synergistic effects on the toxicity and the sublethal effect on larvae. The joint effects on the life-history parameters and the detoxifying enzymes in S. frugiperda, may be useful for implementing IPM programs against this Lepidoptera pest.

MIR337-3p Enhances Mycobacterial Pathogenicity Involving TLR4/MYD88 and STAT3 Signals, Impairing VDR Antimicrobial Response and Fast-Acting Immunity.

Active form of vitamin D (VitD) enhances human innate immunity against Mycobacterium tuberculosis (Mtb) infection. Our previous studies showed that MIR337-3p was highly expressed in lymphocytes of tuberculosis (TB) patients. Here, we identified the mechanism of MIR337-3p in the regulation of fast-acting anti-TB immunity by inhibiting VitD-dependent antimicrobial response pathways. While high-level MIR337-3p expression was induced by mycobacterial infection in cellular models and mice, TB patients exhibited significantly increased MIR337-3p in CD14+ monocytes/macrophages, innate-like Vγ2+ T cells, and CD8+ lymphocytes containing natural killer (NK)/innate lymphoid cells. MIR337-3p promoted the mycobacterial entry/infection and replication/growth in host target cells: macrophages and lung epithelial cells. Such MIR337-3p-enhanced pathogenicity coincided with the MIR337-3p depression of VitD-dependent antimicrobial response of cytochrome P450, family 27, subfamily b, polypeptide 1 (CYP27B1)/Beta-defensin 4 (DEFB4A)/ cathelicidin antimicrobial peptide CAMP pathways. Surprisingly, single MIR337-3p species could specifically target both the Toll-like receptor 4 (TLR4) and signal transducer and activator of transcription 3 (STAT3) 3'-untranslated regions (UTRs) to depress the TLR4/MYD88 and STAT3 signals and impair either of the two signals inhibiting the VitD-dependent antimicrobial pathways in macrophages. Concurrently, human peripheral blood mononuclear cells (PBMCs) expressing high-level MIR337-3p exhibited a reduced ability of innate cell populations to mount fast-acting cellular immunity against intracellular mycobacterial infection. Furthermore, a higher expression of Mir337-3p after mycobacterial infection of mice coincided with much greater colony-forming unit (CFU) counts in lungs and even the death of infected animals, whereas Mir337-3p inhibitor treatment of infected mice reduced Mir337-3p levels and reversed Mir337-3p-mediated increases in CFU counts. Thus, TB-driven single MIR337-3p species could specifically target/impair both TLR4/MYD88 and STAT3 activation signals, inhibiting VitD-dependent antimicrobial response and fast-acting anti-TB immunity, leading to enhanced pathogenicity.

Field-evolved resistance to 11 insecticides and the mechanisms involved in Helicoverpa armigera (Lepidoptera: Noctuidae).

BACKGROUND: To understand the ongoing resistance of cotton bollworm, Helicoverpa armigera, the sensitivity of five field populations to commonly used insecticides, indoxacarb, abamectin, methoxyfenozide, chlorfenapyr, chlorantraniliprole, spinetoram, lambda-cyhalothrin, carbosulfan, metaflumizone, chlorpyrifos, and flufenoxuron, were evaluated. Furthermore, the biochemical and molecular mechanisms of field-evolved resistance in H. armigera were also investigated. RESULTS: Five field populations of H. armigera showed moderate resistance to indoxacarb, chlorantraniliprole, metaflumizone, methoxyfenozide, carbosulfan and lambda-cyhalothrin. The resistance ratio (RR) of indoxacarb was significantly correlated with glutathione-S-transferases (GSTs) activity (r = 0.913, P = 0.011). Methoxyfenozide RR was largely correlated with cytochrome P450s activity (r = 0.860, P = 0.028). Besides, six cytochrome P450s genes of CYP4L5 in AQP, CYP6B7 and CYP9A14 in HDP and BDP, CYP9A17V2 in HDP and YSP, CYP332A1 in HDP, LFP, AQP and YSP, CYP337B1 in YSP, and two GSTs genes of GSTd1 and GSTs1 in HDP were overexpressed (>5-fold). Moreover, indoxacarb RR was positively correlated with the overexpression of GSTs1, GSTd1 and CYP9A14 genes (r = 0.880, 0.98 and 0.86, P = 0.021, 0.001 and 0.028, respectively). The transcript of CYP9A17V2 and CYP337B1 were found to be correlated with metaflumizone RR (r = 0.950, P = 0.004) and carbosulfan RR (r = 0.850, P = 0.033), respectively. CONCLUSION: H. armigera can be effectively controlled using abamectin, chlorfenapyr, chlorpyrifos and spinetoram in Hebei and Shandong provinces. The present study demonstrated that the relative expression level of GSTs1, GSTd1, CYP9A14, CYP9A17V2 and CYP337B1 genes were significantly correlated with the resistance ratio to indoxacarb, metaflumizone and carbosulfan in field H. armigera.

Overexpression of Multiple UDP-Glycosyltransferase Genes Involved in Sulfoxaflor Resistance in Aphis gossypii Glover.

UDP-glycosyltransferases (UGTs) are major phase II enzymes involved in the metabolic detoxification of xenobiotics. In this study, two UGT-inhibitors, 5-nitrouracil and sulfinpyrazone, significantly increased sulfoxaflor toxicity against sulfoxaflor-resistant (Sul-R) Aphis gossypii, whereas there were no synergistic effects in susceptible (Sus) A. gossypii. The activity of UGTs in the Sul-R strain was significantly higher (1.35-fold) than that in the Sus strain. Further, gene expression determination demonstrated that 11 of 23 UGT genes were significantly upregulated (1.40- to 5.46-fold) in the Sul-R strain, among which the expression levels of UGT350A2, UGT351A4, UGT350B2, UGT342C2, and UGT343C2 could be induced by sulfoxaflor. Additionally, knockdown of UGT350A2, UGT351A4, UGT350B2, and UGT343C2 using RNA interference (RNAi) significantly increased sensitivity (1.57- to 1.76-fold) to sulfoxaflor in the Sul-R strain. These results suggested that UGTs might be involved in sulfoxaflor resistance in A. gossypii. These findings will facilitate further work to validate the functional roles of these UGT genes in sulfoxaflor resistance.

Overexpression of ATP-binding cassette transporters associated with sulfoxaflor resistance in Aphis gossypii glover.

BACKGROUND: Sulfoxaflor is a new insecticide for controlling against Aphis gossypii in the field. ATP-binding cassette (ABC) transporters belong to a large superfamily of proteins and play an important role in the detoxification process. However, the potential role of ABC transporters in sulfoxaflor resistance in A. gossypii is unknown. RESULTS: In this study, an ABC transporter inhibitor, verapamil, dramatically increased the toxicity of sulfoxaflor in the resistant population with a synergistic ratio of 8.55. However, verapamil did not synergize sulfoxaflor toxicity in the susceptible population. The contents of ABC transporters were significantly increased in the Sul-R population. Based on RT-qPCR analysis, 10 of 23 ABC transcripts, ABCA1, ABCA2, ABCB1, ABCB5, ABCD1, ABCG7, ABCG16, ABCG26, ABCG27, and MRP7, were up-regulated in the Sul-R population compared to the Sus population. Meanwhile, inductive effects of ABCA1, ABCD1, ABCG7 and ABCG26 by sulfoxaflor were found in A. gossypii. Furthermore, knockdown of ABCA1 and ABCD1 using RNAi significantly increased the sulfoxaflor sensitivity in Sul-R aphids. CONCLUSION: These results suggested that ABC transporters, especially the ABCA1 and ABCD1 genes, might be related with sulfoxaflor resistance in A. gossypii. This study will promote further work to validate the functional roles of these ABCs in sulfoxaflor resistance and might be helpful for the management of sulfoxaflor-resistant A. gossypii.