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Toxoplasmosis is an important zoonotic disease caused by Toxoplasma gondii that can infect almost all warm-blooded animals worldwide, including humans. The high prevalence of T. gondii infection and its ability to cause serious harm to humans and animals, especially immunodeficient individuals, make it a key public health issue. Accurate diagnostic tools with high sensitivity are needed for controlling T. gondii infection. In the current study, we compared the performance of recombinant SAG2, GRA6, and GRA7 in ELISA for the serological diagnosis of T. gondii infection in cats. We further investigated the antigenicity of recombinant dense granule protein 3 (rGRA3), rGRA5, rGRA8, and rSRS29A expressed in a plant-based, cell-free expression system for detecting antibodies in T. gondii-infected cats. In summary, our data suggest that GRA7 is more sensitive than the other two antigens for the serodiagnosis of T. gondii infection in cats, and GRA3 expressed in the cell-free system is also a priming antigen in serological tests for detecting T. gondii infection in cats.
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BACKGROUND: Dysregulation of vascular smooth muscle cell (VSMC) function leads to a variety of diseases such as atherosclerosis and hyperplasia after injury. However, antiproliferative drug targeting VSMC exhibits poor specificity. Therefore, there is an urgent to develop highly specific antiproliferative drugs to prevention and treatment VSMC dedifferentiation associated arteriosclerosis. Kanglexin (KLX), a new anthraquinone compound designed by our team, has potential to regulate VSMC phenotype according to the physicochemical properties. PURPOSE: This project aims to evaluate the therapeutic role of KLX in VSMC dedifferentiation and atherosclerosis, neointimal formation and illustrates the underlying molecular mechanism. METHODS: In vivo, the ApoE-/- mice were fed with high-fat diet (HFD) for a duration of 13 weeks to establish the atherosclerotic model. And rat carotid artery injury model was performed to establish the neointimal formation model. In vitro, PDGF-BB was used to induce VSMC dedifferentiation. RESULTS: We found that KLX ameliorated the atherosclerotic progression including atherosclerotic lesion formation, lipid deposition and collagen deposition in aorta and aortic sinus in atherosclerotic mouse model. In addition, The administration of KLX effectively ameliorated neointimal formation in the carotid artery following balloon injury in SD rats. The findings derived from molecular docking and surface plasmon resonance (SPR) experiments unequivocally demonstrate that KLX had potential to bind PDGFR-ß. Mechanism research work proved that KLX prevented VSMC proliferation, migration and dedifferentiation via activating the PDGFR-ß-MEK -ERK-ELK-1/KLF4 signaling pathway. CONCLUSION: Collectively, we demonstrated that KLX effectively attenuated the progression of atherosclerosis in ApoE-/- mice and carotid arterial neointimal formation in SD rats by inhibiting VSMC phenotypic conversion via PDGFR-ß-MEK-ERK-ELK-1/KLF4 signaling. KLX exhibits promising potential as a viable therapeutic agent for the treatment of VSMC phenotype conversion associated arteriosclerosis.
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Antraquinonas , Desdiferenciación Celular , Factor 4 Similar a Kruppel , Músculo Liso Vascular , Neointima , Animales , Masculino , Ratones , Ratas , Antraquinonas/farmacología , Arteriosclerosis/tratamiento farmacológico , Arteriosclerosis/prevención & control , Aterosclerosis/tratamiento farmacológico , Becaplermina/farmacología , Traumatismos de las Arterias Carótidas/tratamiento farmacológico , Desdiferenciación Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Dieta Alta en Grasa , Modelos Animales de Enfermedad , Factores de Transcripción de Tipo Kruppel/metabolismo , Ratones Endogámicos C57BL , Simulación del Acoplamiento Molecular , Músculo Liso Vascular/efectos de los fármacos , Miocitos del Músculo Liso/efectos de los fármacos , Neointima/tratamiento farmacológico , Ratas Sprague-Dawley , Receptores del Factor de Crecimiento Derivado de Plaquetas/metabolismo , Transducción de Señal/efectos de los fármacosRESUMEN
Introduction: Reverse genetics has become an indispensable tool to gain insight into the pathogenesis of viruses and the development of vaccines. The yeast-based synthetic genomics platform has demonstrated the novel capabilities to genetically reconstruct different viruses. Methods: In this study, a transformation-associated recombination (TAR) system in yeast was used to rapidly rescue different strains of feline infectious peritonitis virus, which causes a deadly disease of cats for which there is no effective vaccine. Results and discussion: Using this system, the viruses could be rescued rapidly and stably without multiple cloning steps. Considering its speed and ease of manipulation in virus genome assembly, the reverse genetics system developed in this study will facilitate the research of the feline coronaviruses pathogenetic mechanism and the vaccine development.
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Psoriasis is a chronic inflammatory disease whose etiology is directly related to the dysregulation of cutaneous immune homeostasis. However, how to finely modulate the skin immune microenvironment to restore homeostasis remains an important challenge. Inspired by the natural attribute of tumor exosomes in the immune escape, the tumor-derived exosomes as an active targeting nanoplatform for the effective treatment of inflammatory skin disorder were first reported. As keratinocytes and immune cells express high PD-1 during the onset of psoriasiform skin inflammation, the PD-L1-positive exosomes derived from melanoma cells carrying pristimerin with extremely anti-inflammatory potential were yielded to treat psoriasis. The PD-L1+ exosomes carrying pristimerin were characterized, and the cellular uptake was performed to evaluate the PD-1 target capability. The anti-inflammatory action of PD-L1+ exosomes carrying pristimerin was observed in both in vitro and in vivo models of psoriasis. Our exosomes substantially increased pristimerin uptake with CD4+ T cells and keratinocytes, significantly inhibited the proliferation of Th17 cells, and promoted Treg differentiation in a psoriasis-like model. Obviously, PD-L1+ exosomes carrying pristimerin significantly and safely reversed imiquimod (IMQ)-induced psoriasis in mice, indicated by reducing epidermal thickness, decreasing plaque formation, and suppressed excessive inflammatory response, due to its dual targeting of both CD4+ T cells and keratinocytes gathering around the lesion. The inflammatory cell infiltration and pro-inflammatory cytokine production in psoriasis were suppressed by our engineered exosomes. Besides, PD-L1+ exosomes carrying pristimerin treatment alleviated ferroptosis-related changes in psoriatic skin, thereby dampening excessive inflammation and, in turn, decreasing the abnormal proliferation of keratinocytes in psoriatic lesions. This study demonstrates that our engineered exosomes can not only act as a treat-to-target strategy for psoriasis treatment but also provide insight in clinical application of inflammatory disorders.
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Exocytosis is a key active process in cells by which proteins are released in bulk via the fusion of exocytic vesicles with the plasma membrane. Soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) protein-mediated vesicle fusion with the plasma membrane is essential in most exocytotic pathways. In mammalian cells, the vesicular fusion step of exocytosis is normally mediated by Syntaxin-1 (Stx1) and SNAP25 family proteins (SNAP25 and SNAP23). However, in Toxoplasma gondii, a model organism of Apicomplexa, the only SNAP25 family protein, with a SNAP29-like molecular structure, is involved in vesicular fusion at the apicoplast. Here, we reveal that an unconventional SNARE complex comprising TgStx1, TgStx20, and TgStx21 mediates vesicular fusion at the plasma membrane. This complex is essential for the exocytosis of surface proteins and vesicular fusion at the apical annuli in T. gondii.
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Toxoplasma , Animales , Toxoplasma/metabolismo , Proteínas de Transporte Vesicular/metabolismo , Membrana Celular/metabolismo , Exocitosis , Fusión de Membrana , Proteínas Qa-SNARE/genética , Proteínas Qa-SNARE/metabolismo , MamíferosRESUMEN
Psoriasis is a chronic inflammatory skin disease. Inflammation and oxidative stress play crucial roles in the pathogenesis of psoriasis. Cannabinoid receptor type 2 (CB2R) is an attractive target for treating various inflammatory disorders. However, the precise role and mechanism of CB2R activation in psoriasis remain to be further elucidated. In this study, imiquimod (IMQ)-induced experimental psoriasis mice and tumor necrosis factor-α (TNF-α)-activated keratinocytes (HaCaT) were used to examine the effect of CB2R activation on psoriasis-like lesions and the mechanism in vivo and in vitro. Our results demonstrated that activation of CB2R by the specific agonist GW842166X (GW) significantly ameliorated IMQ-induced psoriasiform skin lesions in mice by reducing epidermal thickness and decreasing plaque thickness. On the one hand, GW alleviated inflammation by decreasing inflammatory cytokines and abating inflammatory cell infiltration. On the other hand, this treatment reduced the level of iNOS and downregulated the expression of CB2R in psoriatic skin tissue. Further studies suggested that the Kelch-like ECH-associated protein 1/nuclear factor erythroid-2-related factor (Keap1/Nrf2) signaling pathway might be involved. Our findings reveal that selective activation of CB2R may serve as a new strategy for the treatment of psoriasis.
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Psoriasis , Enfermedades de la Piel , Animales , Ratones , Proteína 1 Asociada A ECH Tipo Kelch/metabolismo , Factor 2 Relacionado con NF-E2/metabolismo , Psoriasis/inducido químicamente , Psoriasis/tratamiento farmacológico , Psoriasis/metabolismo , Enfermedades de la Piel/patología , Piel/metabolismo , Queratinocitos/metabolismo , Inflamación/metabolismo , Imiquimod/efectos adversos , Imiquimod/metabolismo , Citocinas/metabolismo , Estrés Oxidativo , Modelos Animales de Enfermedad , Ratones Endogámicos BALB CRESUMEN
OBJECTIVE: This experiment was designed to demonstrate Mesenchymal stem cells (MSCs) derived from kidney can alleviate cisplatin-induced kidney injury and renal cell apoptosis through paracrine pathway. METHODS: Firstly, MSCs were isolated from kidney of young rats, and their surface-specific markers were identified by Reverse Transcription-Polymerase Chain Reaction (RT-PCR) and immunofluorescence staining. Self-renewal ability of Kidney Mesenchymal Stem Cells (KMSCs) was observed by cell counting and 5-Bromo-2'-deoxyuridine (BrdU) fluorescence staining. KMSCs at logarithmic growth stage were traced and injected into rat through tail vein. RESULTS: The results showed that KMSCs homed in the kidney tissues, decreased the secretion of inflammatory factors (CRP, TNFα, IL-1ß, IL-6), and alleviated renal function. Hematoxylin and Eosin (Hï¼E), Masson and Periodic Acid-silver Methenamine (PASM) staining showed that KMSCs could alleviate pathological damage in rats. Terminal Deoxynucleotidyl Transferase mediated dUTP Nick-End Labeling (TUNEL) assay showed that KMSCs could reduce the apoptosis of rat kidney cells induced by cisplatin. Finally, Immunohistochemistry (IHC) results showed that cisplatin could induce higher expression of the pro-apoptotic protein Bax and lower expression of anti-apoptotic Bcl-2 in kidney tissues. However, KMSCs could reverse the pro-apoptotic effect of cisplatin on kidney cells and improve the survival rate of rats. CONCLUSIONS: In conclusion, KMSCs were successfully isolated from kidney tissues, and KMSCs have therapeutic effects on rat kidney injury induced by cisplatin.
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Cisplatino , Células Madre Mesenquimatosas , Ratas , Animales , Cisplatino/toxicidad , Ratas Sprague-Dawley , Riñón/metabolismo , Apoptosis , Células Madre Mesenquimatosas/metabolismoRESUMEN
The efficacy of immunotherapy combined with other therapeutic modalities in the management of cancer has been extensively studied. However, no effective strategy to improve the antitumor effects of immunotherapy at the tumor site has been developed. In this study, we describe a nanoformulation (CP) that integrates ferroptosis-inducing cannabinoid nanoparticles with immunostimulatory Poly(I:C) to enhance antitumor immune responses by activating ferroptosis-immunotherapy pathways. The results indicated that CP nanoformulation effectively induced ferroptosis, cellular immunogenic death, and anti-tumor immune responses which initiate T cell responses leading to the inhibition of established tumors. In addition, CP nanoformulations reversed the tumor immunosuppressive microenvironment and promoted tumor ferroptosis. These results indicated that the self-amplifying nanoformulation may be an effective strategy for broad-spectrum cancer immunotherapy.
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Ferroptosis , Neoplasias , Humanos , Terapia de Inmunosupresión , Inmunoterapia , Microambiente Tumoral , Inmunosupresores , Línea Celular TumoralRESUMEN
In apicomplexan parasites, the macroautophagy/autophagy machinery is repurposed to maintain the plastid-like organelle apicoplast. Previously, we showed that in Toxoplasma and Plasmodium, ATG12 interacts with ATG5 in a non-covalent manner, in contrast to the covalent interaction in most organisms. However, it remained unknown whether apicomplexan parasites have functional orthologs of ATG16L1, a protein that is essential for the function of the covalent ATG12-ATG5 complex in vivo in other organisms. Furthermore, the mechanism used by the autophagy machinery to maintain the apicoplast is unclear. We report that the ATG12-ATG5-ATG16L complex exists in Toxoplasma gondii (Tg). This complex is localized on isolated structures at the periphery of the apicoplast dependent on TgATG16L. Inducible depletion of TgATG12, TgATG5, or TgATG16L caused loss of the apicoplast and affected parasite growth. We found that a putative soluble N-ethylmaleimide sensitive factor attachment protein receptor (SNARE) protein, synaptosomal-associated protein 29 (TgSNAP29, Qbc SNARE), is required to maintain the apicoplast in T. gondii. TgSNAP29 depletion disrupted TgATG8 localization at the apicoplast. Additionally, we identified a putative ubiquitin-interacting motif-docking site (UDS) of TgATG8. Mutation of the UDS site abolished TgATG8 localization on the apicoplast but not lipidation. These findings suggest that the TgATG12-TgATG5-TgATG16L complex is required for biogenesis of the apicoplast, in which TgATG8 is translocated to the apicoplast via vesicles in a SNARE -dependent manner in T. gondii.Abbreviations: AID: auxin-inducible degron; CCD: coiled-coil domain; HFF: human foreskin fibroblast; IAA: indole-3-acetic acid; LAP: LC3-associated phagocytosis; NAA: 1-naphthaleneacetic acid; PtdIns3P: phosphatidylinositol-3-phosphate; SNARE: soluble N-ethylmaleimide sensitive factor attachment protein receptor; UDS: ubiquitin-interacting motif-docking site; UIM: ubiquitin-interacting motif.
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Apicoplastos , Parásitos , Toxoplasma , Animales , Humanos , Toxoplasma/genética , Toxoplasma/metabolismo , Apicoplastos/genética , Apicoplastos/metabolismo , Etilmaleimida/metabolismo , Autofagia , Ubiquitinas/metabolismo , Proteínas Protozoarias/genética , Proteína 12 Relacionada con la Autofagia/metabolismo , Proteínas Qb-SNARE/metabolismo , Proteínas Qc-SNARE , Proteína 5 Relacionada con la Autofagia/metabolismoRESUMEN
BACKGROUND: Allergic rhinitis (AR) is a common disease worldwide. Imbalances in T helper (Th) cell differentiation and the dysregulation of related cytokines form the immunological basis of AR. miR-126 may play an important regulatory role in AR as a new marker and predictor of the disease. Therefore, the aim of this study was to explore the regulatory effects of miR-126 on Th cell differentiation and cytokine expression in AR. METHODS: T lymphocytes and rat models were transfected with a miR-126 mimic and an inhibitor. The expression of miR-126 and Th cell-related cytokines was detected by RT-qPCR and western blotting. The serum IgE levels were detected using ELISA. In the nasal mucosa, pathological changes were observed by HE staining, protein expression was detected by immunohistochemistry, and the differentiation ratio of Th cell subsets was detected by flow cytometry. RESULTS: During the occurrence and development of AR, the expression of miR-126 and the IgE levels were increased in the AR group. The number of Treg cell subsets decreased in the AR rats, increased after the miR-126 agomir intervention and decreased after miR-126 antagomir intervention. The number of Th1 and Th2 cell subsets increased in the AR rats, decreased after miR-126 agomir intervention and increased after the miR-126 antagomir intervention. CONCLUSION: We propose that miR-126 may be involved in the pathogenesis of AR by positively regulating the expression of Treg cytokines and negatively regulating the expression of the Th1 and Th2 cytokines.
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MicroARNs , Rinitis Alérgica , Animales , Antagomirs/metabolismo , Diferenciación Celular , Citocinas/metabolismo , Modelos Animales de Enfermedad , Inmunoglobulina E/metabolismo , Inmunoglobulina E/farmacología , MicroARNs/genética , MicroARNs/metabolismo , Ratas , Rinitis Alérgica/metabolismo , Linfocitos T ReguladoresRESUMEN
Toxoplasma gondii is a worldwide distributed zoonotic pathogen that threatens public health. However, there have been limited data for T. gondii infection in wild rats (Rattus norvegicus) in China. In the present study, a total of 227 wild rats were captured from three mink farms to investigate T. gondii infection in Shandong Province, eastern China. The DNA was extracted from 25 mg rats' brain tissues and subjected to a PCR amplification by targeting to the T. gondii B1. In 227 wild rat samples, 18 samples (7.93%) were positive for T. gondii. Then, the positive samples were further genotyped based on eight genetic markers, including eight nuclear loci (SAG1, 5'-SAG2, and 3'-SAG2, alternative SAG2, SAG3, GRA6, c29-2, and L358) and an apicoplast locus (Apico) by using the multilocus PCR-restriction fragment length polymorphism technology. Of these samples, eight were genotyped at nine nuclear loci, and two were genotyped at eight nuclear loci, forming three known genotypes (ToxoDB no. 43, ToxoDB no. 91, and ToxoDB no. 189) and two new genotypes. The closest ToxoDB genotypes were observed in wild rats, suggesting the differences in the population structure of the T. gondii between breed farm animals and wild rats. These data revealed the genetic variability of T. gondii in wild rats on mink farms in Shandong Province, with possible implication for public health.
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Enfermedades de los Roedores , Toxoplasma , Toxoplasmosis Animal , Animales , China/epidemiología , ADN Protozoario/genética , Granjas , Genotipo , Visón , Polimorfismo de Longitud del Fragmento de Restricción , Ratas , Enfermedades de los Roedores/epidemiología , Toxoplasma/genética , Toxoplasmosis Animal/diagnóstico , Toxoplasmosis Animal/epidemiologíaRESUMEN
The immunoproteasomes are specific proteasomes that clear oxidant-damaged proteins under inflammatory conditions in various diseases. Toxoplasma gondii (T. gondii) infects the central nervous system and causeencephalitis. However, the relationship between the immunoproteasomes and brain inflammation during T. gondii infection is not well characterized. In this study, we established an in vivo mouse model of T. gondii PLK strain infection via intraperitoneal injection and evaluated the expression of immunoproteasome subunits in the brains of infected mice. The results demonstrated that first, pathological changes in the brains of infected mice increase in severity over time. Second, following T. gondii infection, activated microglia and astrocytes undergo a series of functional alterations and morphological transformations, including proliferation and migration. Third, T. gondii infection induces expression of inflammatory cytokines, including IFN-γ, IL-1ß, TNF-α, and IL-6. Fourth, the immunoproteasome subunits low-molecular-weight polypeptide 2 (LMP2), LMP7, and LMP10 mRNA and protein levels are significantly upregulated in T. gondii-infected mouse brains, as shown by RT-qPCR and western blot analysis, compared with that in vehicle-treated brains, and their expression is localized in the microglia, astrocytes, and neurons of T. gondii-infected brains, as determined via immunofluorescence staining. Furthermore, the western blot mean gray value for the immunoproteasome subunits and the positive microglia and astrocyte immunohistochemical signals in the brains of T. gondii-infected mice were positively correlated, indicating that the observed relationships were highly significant. Therefore, it was concluded that the induction of the immunoproteasomes is a pathogenic mechanism underlying T. gondii infection-induced inflammation.
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Inflamación/genética , Complejo de la Endopetidasa Proteasomal/genética , Toxoplasmosis/genética , Animales , Astrocitos/metabolismo , Astrocitos/patología , Encéfalo/metabolismo , Encéfalo/microbiología , Encéfalo/patología , Humanos , Inflamación/microbiología , Inflamación/patología , Interferón gamma/genética , Interleucina-1beta/genética , Interleucina-6/genética , Ratones , Microglía/metabolismo , Microglía/patología , Neuronas/metabolismo , Neuronas/patología , Toxoplasma/genética , Toxoplasma/patogenicidad , Toxoplasmosis/microbiología , Toxoplasmosis/patología , Factor de Necrosis Tumoral alfa/genéticaRESUMEN
Radiation-induced lung injury (RILI) is a potentially fatal and dose-limiting complication of thoracic cancer radiotherapy. However, effective therapeutic agents for this condition are limited. Here, we describe a novel strategy to exert additive effects of a non-erythropoietic EPO derivative (ARA290), along with a free radical scavenger, superoxide dismutase (SOD), using a bioengineered nanoreactor (SOD@ARA290-HBc). ARA290-chimeric nanoreactor makes SOD present in a confined reaction space by encapsulation into its interior to heighten stability against denaturing stimuli. In a RILI mouse model, intratracheal administration of SOD@ARA290-HBc was shown to significantly ameliorate acute radiation pneumonitis and pulmonary fibrosis. Our investigations revealed that SOD@ARA290-HBc performs its radioprotective effects by protecting against radiation induced alveolar epithelial cell apoptosis and ferroptosis, suppressing oxidative stress, inhibiting inflammation and by modulating the infiltrated macrophage phenotype, or through a combination of these mechanisms. In conclusion, SOD@ARA29-HBc is a potential therapeutic agent for RILI, and given its multifaceted roles, it may be further developed as a translational nanomedicine for other related disorders.
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Lesión Pulmonar , Fibrosis Pulmonar , Traumatismos por Radiación , Animales , Pulmón , Ratones , NanotecnologíaRESUMEN
Vesicular trafficking is a fundamental cellular process involved in material transport in eukaryotes, but the diversity of the intracellular compartments has prevented researchers from obtaining a clear understanding of the specific functions of vesicular trafficking factors, including SNAREs, tethers, and Rab GTPases, in Apicomplexa. In this study, we analyzed the localization of SNAREs and investigated their roles in vesicular trafficking in Toxoplasma gondii. Our results revealed the specific localizations of SNAREs in the endoplasmic reticulum (ER) (T. gondii Stx18 [TgStx18] and TgStx19), Golgi stacks (TgGS27), and endosome-like compartment (TgStx10 and TgStx12). The conditional ablation of ER- and Golgi-residing SNAREs caused severe defects in the secretory system. Most importantly, we found an R-SNARE (TgVAMP4-2) that is targeted to the apicoplast; to our knowledge, this work provides the first information showing a SNARE protein on endosymbiotic organelles and functioning in vesicular trafficking in eukaryotes. Conditional knockout of TgVAMP4-2 blocked the entrance of TgCPN60, TgACP, TgATrx2, and TgATrx1 into the apicoplast and interfered with the targeting of TgAPT1 and TgFtsH1 to the outermost membrane of the apicoplast. Together, our findings revealed the functions of SNAREs in the secretory system and the transport of nucleus-encoded proteins to an endosymbiotic organelle in a model organism of Apicomplexa. IMPORTANCE SNAREs are essential for the fusion of the transport vesicles and target membranes and, thus, provide perfect targets for obtaining a global view of the vesicle transport system. In this study, we report that a novel Qc-SNARE (TgStx19) instead of Use1 is located at the ER and acts as a partner of TgStx18 in T. gondii. TgGS27 and the tethering complex TRAPP III are conserved and critical for the biogenesis of the Golgi complex in T. gondii. A novel R-SNARE, TgVAMP4-2, is found on the outermost membrane of the apicoplast. The transport of NEAT proteins into the secondary endosymbiotic organelle depends on its function. To our knowledge, this work provides the first mention of a SNARE located on endosymbiotic organelles that functions in vesicular trafficking in eukaryotes.
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Apicoplastos/fisiología , Proteínas Protozoarias/metabolismo , Proteínas SNARE/metabolismo , Toxoplasma/metabolismo , Retículo Endoplásmico/metabolismo , Aparato de Golgi/metabolismo , Biogénesis de Organelos , Transporte de Proteínas , Proteínas Protozoarias/genética , Proteínas SNARE/genética , Toxoplasma/genéticaRESUMEN
Metastasis is the leading cause of death in cancer patients. Eliciting anti-tumor immune responses against lung metastasis is hindered by the immunosuppressive microenvironment. This study explored a biomimetic nanoformulation, comprising a nanovaccine (OP) that delivers tumor antigens and adjuvants spatially and temporally in a virus-like manner, and a pulmonary surfactant-biomimetic liposome with an immunomodulator, JQ1 (PS-JQ1). The findings of this study showed that intratracheal administration of OP+PS-JQ1 activated lung immune cells without concomitant excess inflammation, enhanced tumor antigen cross-presentation, generated a significantly high antigen-specific CD8+ T cell response, and reshaped the immunocellular composition in B16 melanoma tumor-bearing lung. OP+PS-JQ1 nanoformulation exhibited a striking immunotherapeutic efficacy, induced local and systemic tumor suppression, improved survival of mice, initiated immune memory that prevents recurrence of secondary tumors. This stable and nontoxic nanoformulation provides a simple, flexible, and robust strategy for augmenting anti-tumor immunity for metastatic cancer. STATEMENT OF SIGNIFICANCE: Egg glue proteins are produced by female insects, which can make the eggs firmly attached to the oviposition sites, not affected by wind and rain. However, genes encoding insect egg glue proteins have not yet been reported, and the molecular mechanism underpinning their adhesion is still unknown. Our study makes a significant contribution to the literature as it identifies the sequence, structure, adhesive property, and mechanism of silkworm egg glue protein. Furthermore, it outlines key insights into the structure-function relationships associated with egg glue proteins. We believe that this paper will be of interest to the readership of your journal as it identifies the first complete sequence of insect egg glue proteins, thereby highlighting their potentials future applications in both the biomedical and technical fields.
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Biomimética , Recurrencia Local de Neoplasia , Adyuvantes Inmunológicos/farmacología , Animales , Antígenos de Neoplasias , Linfocitos T CD8-positivos , Femenino , Humanos , Inmunoterapia , Ratones , Ratones Endogámicos C57BL , Microambiente TumoralRESUMEN
The apicoplast is a non-photosynthetic relict plastid of Apicomplexa that evolved from a secondary symbiotic system. During its evolution, most of the genes derived from its alga ancestor were lost. Only genes involved in several valuable metabolic pathways, such as the synthesis of isoprenoid precursors, heme, and fatty acids, have been transferred to the host genome and retained to help these parasites adapt to a complex life cycle and various living environments. The biological function of an apicoplast is essential for most apicomplexan parasites. Considering their potential as drug targets, the metabolic functions of this symbiotic organelle have been intensively investigated through computational and biological means. Moreover, we know that not only organellar metabolic functions are linked with other organelles, but also their biogenesis processes have developed and evolved to tailor their biological functions and proper inheritance. Several distinct features have been found in the biogenesis process of apicoplasts. For example, the apicoplast borrows a dynamin-related protein (DrpA) from its host to implement organelle division. The autophagy system has also been repurposed for linking the apicoplast and centrosome during replication and the division process. However, many vital questions remain to be answered about how these parasites maintain and properly inherit this symbiotic organelle. Here we review our current knowledge about its biogenesis process and discuss several critical questions remaining to be answered in this field.
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Apicomplexa/fisiología , Apicoplastos/fisiología , Biogénesis de OrganelosRESUMEN
Methionine aminopeptidases (MAPs), which remove the N-terminal methionine from newly synthesised proteins, are present in all life forms. Three type I MAPs and one type II MAP are encoded in the genome of Toxoplasma gondii. In this study, we found that the inducible knockdown of each type I TgMAP (TgMAP1a-c) reduced the growth and proliferation of the parasite significantly. Among them, TgMAP1c was found to be localised to the apicoplast of the parasite. Inducible knockdown of TgMAP1c led to a defect in the abundance of apicoplast-encoded transcripts, and a later reduction in the apicoplast genome copy number and loss of the apicoplast structure. This finding indicates that transcription of the apicoplast genome was impaired upon knockdown of TgMAP1c. We also found that the function of TgMAP1c in apicoplast biogenesis depends on its enzymatic domain. Expression of a recombinant protein in which the active domain of TgMAP1c was replaced with that of TgMAP1a or TgMAP1b could not restore the defective growth and replication phenotype caused by knockdown of TgMAP1c, indicating that these three enzymes have distinct substrate preferences. An in vitro analysis also revealed that TgMAP1c is an active enzyme that acts specifically on the substrate H-Met-p-NA. In addition, inducible knockdown of TgMAP1c reduced the virulence of T. gondii in mice. Taken together, these results demonstrate that TgMAP1c plays a key role in the biogenesis and maintenance of the T. gondii apicoplast.
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Apicoplastos , Biogénesis de Organelos , Proteínas Protozoarias/genética , Toxoplasma , Animales , Apicoplastos/genética , Técnicas de Silenciamiento del Gen , Ratones , Toxoplasma/genéticaRESUMEN
Gamma interferon (IFN-γ)-induced innate immune responses play important roles in the inhibition of Toxoplasma gondii infection. It has been reported that IFN-γ stimulates non-acidification-dependent growth restriction of T. gondii in HeLa cells, but the mechanism remains unclear. Here, we found that γ-aminobutyric acid (GABA) receptor-associated protein-like 2 (GABARAPL2) plays a critical role in parasite restriction in IFN-γ-treated HeLa cells. GABARAPL2 is recruited to membrane structures surrounding parasitophorous vacuoles (PV). Autophagy adaptors are required for the proper localization and function of GABARAPL2 in the IFN-γ -induced immune response. These findings provide further understanding of a noncanonical autophagy pathway responsible for IFN-γ-dependent inhibition of T. gondii growth in human HeLa cells and demonstrate the critical role of GABARAPL2 in this response.