RESUMEN

Molecular-profiling-based cancer diagnosis has significant implications for predicting disease prognosis and selecting targeted therapeutic interventions. The analysis of cancer-derived extracellular vesicles (EVs) provides a noninvasive and sequential method to assess the molecular landscape of cancer. Here, we developed an all-in-one fusogenic nanoreactor (FNR) encapsulating DNA-fueled molecular machines (DMMs) for the rapid and direct detection of EV-associated microRNAs (EV miRNAs) in a single step. This platform was strategically designed to interact selectively with EVs and induce membrane fusion under a specific trigger. After fusion, the DMMs recognized the target miRNA and initiated nonenzymatic signal amplification within a well-defined reaction volume, thus producing an amplified fluorescent signal within 30 min. We used the FNRs to analyze the unique expression levels of three EV miRNAs in various biofluids, including cell culture, urine, and plasma, and obtained an accuracy of 86.7% in the classification of three major breast cancer (BC) cell lines and a diagnostic accuracy of 86.4% in the distinction between patients with cancer and healthy donors. Notably, a linear discriminant analysis revealed that increasing the number of miRNAs from one to three improved the accuracy of BC patient discrimination from 78.8 to 95.4%. Therefore, this all-in-one diagnostic platform performs nondestructive EV processing and signal amplification in one step, providing a straightforward, accurate, and effective individual EV miRNA analysis strategy for personalized BC treatment.

RESUMEN

BACKGROUND: Pancreatic ductal adenocarcinoma (PDAC) is an aggressive malignancy with a poor survival rate, largely due to the lack of early diagnosis. Although myeloid cells are crucial in the tumour microenvironment, whether their specific subset can be a biomarker of PDAC progression is unclear. METHODS: We analysed IL-22 receptor expression in PDAC and peripheral blood. Additionally, we analysed gene expression profiles of IL-10R2+/IL-22R1+ myeloid cells and the presence of these cells using single-cell RNA sequencing and murine orthotropic PDAC models, respectively, followed by examining the immunosuppressive function of IL-10R2+/IL-22R1+ myeloid cells. Finally, the correlation between IL-10R2 expression and PDAC progression was evaluated. RESULTS: IL-10R2+/IL-22R1+ myeloid cells were present in PDAC and peripheral blood. Blood IL-10R2+ myeloid cells displayed a gene expression signature associated with tumour-educated circulating monocytes. IL-10R2+/IL-22R1+ myeloid cells from human myeloid cell culture inhibited T cell proliferation. By mouse models for PDAC, we found a positive correlation between pancreatic tumour growth and increased blood IL-10R2+/IL-22R1+ myeloid cells. IL-10R2+/IL-22R1+ myeloid cells from an early phase of the PDAC model suppressed T cell proliferation and cytotoxicity. IL-10R2+ myeloid cells indicated tumour recurrence 130 days sooner than CA19-9 in post-pancreatectomy patients. CONCLUSIONS: IL-10R2+/IL-22R1+ myeloid cells in the peripheral blood might be an early marker of PDAC prognosis.

Asunto(s)

Biomarcadores de Tumor , Carcinoma Ductal Pancreático , Subunidad beta del Receptor de Interleucina-10 , Células Mieloides , Recurrencia Local de Neoplasia , Neoplasias Pancreáticas , Receptores de Interleucina , Carcinoma Ductal Pancreático/genética , Carcinoma Ductal Pancreático/patología , Carcinoma Ductal Pancreático/diagnóstico , Carcinoma Ductal Pancreático/sangre , Humanos , Animales , Neoplasias Pancreáticas/patología , Neoplasias Pancreáticas/genética , Neoplasias Pancreáticas/diagnóstico , Neoplasias Pancreáticas/sangre , Ratones , Biomarcadores de Tumor/genética , Biomarcadores de Tumor/sangre , Receptores de Interleucina/genética , Células Mieloides/metabolismo , Células Mieloides/patología , Recurrencia Local de Neoplasia/genética , Recurrencia Local de Neoplasia/patología , Subunidad beta del Receptor de Interleucina-10/genética , Femenino , Masculino , Microambiente Tumoral/genética , Línea Celular Tumoral

RESUMEN

BACKGROUND: Patients with fibro-calcific aortic valve disease (FCAVD) have lipid depositions in their aortic valve that engender a proinflammatory impetus toward fibrosis and calcification and ultimately valve leaflet stenosis. Although the lipoprotein(a)-autotaxin (ATX)-lysophosphatidic acid axis has been suggested as a potential therapeutic target to prevent the development of FCAVD, supportive evidence using ATX inhibitors is lacking. We here evaluated the therapeutic potency of an ATX inhibitor to attenuate valvular calcification in the FCAVD animal models. METHODS: ATX level and activity in healthy participants and patients with FCAVD were analyzed using a bioinformatics approach using the Gene Expression Omnibus datasets, enzyme-linked immunosorbent assay (ELISA), immunohistochemistry, and western blotting. To evaluate the efficacy of ATX inhibitor, interleukin-1 receptor antagonist-deficient (Il1rn-/-) mice and cholesterol-enriched diet-induced rabbits were used as the FCAVD models, and primary human valvular interstitial cells (VICs) from patients with calcification were employed. RESULTS: The global gene expression profiles of the aortic valve tissue of patients with severe FCAVD demonstrated that ATX gene expression was significantly upregulated and correlated with lipid retention (r = 0.96) or fibro-calcific remodeling-related genes (r = 0.77) in comparison to age-matched non-FCAVD controls. Orally available ATX inhibitor, BBT-877, markedly ameliorated the osteogenic differentiation and further mineralization of primary human VICs in vitro. Additionally, ATX inhibition significantly attenuated fibrosis-related factors' production, with a detectable reduction of osteogenesis-related factors, in human VICs. Mechanistically, ATX inhibitor prohibited fibrotic changes in human VICs via both canonical and non-canonical TGF-ß signaling, and subsequent induction of CTGF, a key factor in tissue fibrosis. In the in vivo FCAVD model system, ATX inhibitor exposure markedly reduced calcific lesion formation in interleukin-1 receptor antagonist-deficient mice (Il1rn-/-, P = 0.0210). This inhibition ameliorated the rate of change in the aortic valve area (P = 0.0287) and mean pressure gradient (P = 0.0249) in the FCAVD rabbit model. Moreover, transaortic maximal velocity (Vmax) was diminished with ATX inhibitor administration (mean Vmax = 1.082) compared to vehicle control (mean Vmax = 1.508, P = 0.0221). Importantly, ATX inhibitor administration suppressed the effects of a high-cholesterol diet and vitamin D2-driven fibrosis, in association with a reduction in macrophage infiltration and calcific deposition, in the aortic valves of this rabbit model. CONCLUSIONS: ATX inhibition attenuates the development of FCAVD while protecting against fibrosis and calcification in VICs, suggesting the potential of using ATX inhibitors to treat FCAVD.

Asunto(s)

Estenosis de la Válvula Aórtica , Válvula Aórtica/patología , Calcinosis , Humanos , Animales , Ratones , Conejos , Estenosis de la Válvula Aórtica/tratamiento farmacológico , Osteogénesis , Calcinosis/tratamiento farmacológico , Células Cultivadas , Fibrosis , Colesterol , Receptores de Interleucina-1 , Lípidos

RESUMEN

BACKGROUND: Toll-like receptor 4 (TLR4) conducts a highly regulated inflammatory process by limiting the extent of inflammation to avoid toxicity and tissue damage, even in bone tissues. Thus, it is plausible that strategies for the maintenance of normal bone-immunity to prevent undesirable bone damage by TLR4 activation can exist, but direct evidence is still lacking. METHODS: Osteoclast precursors (OCPs) obtained from WT or Slit3-deficient mice were differentiated into osteoclast (OC) with macrophage colony-stimulating factor (M-CSF), RANK ligand (RANKL) and lipopolysaccharide (LPS) by determining the number of TRAP-positive multinuclear cells (TRAP+ MNCs). To determine the alteration of OCPs population, fluorescence-activated cell sorting (FACS) was conducted in bone marrow cells in mice after LPS injection. The severity of bone loss in LPS injected WT or Slit3-deficient mice was evaluated by micro-CT analysis. RESULT: We demonstrate that TLR4 activation by LPS inhibits OC commitment by inducing the concomitant expression of miR-218-2-3p and its host gene, Slit3, in mouse OCPs. TLR4 activation by LPS induced SLIT3 and its receptor ROBO1 in BMMs, and this SLIT3-ROBO1 axis hinders RANKL-induced OC differentiation by switching the protein levels of C/EBP-ß isoforms. A deficiency of SLIT3 resulted in increased RANKL-induced OC differentiation, and the elevated expression of OC marker genes including Pu.1, Nfatc1, and Ctsk. Notably, Slit3-deficient mice showed expanded OCP populations in the bone marrow. We also found that miR-218-2 was concomitantly induced with SLIT3 expression after LPS treatment, and that this miRNA directly suppressed Tnfrsf11a (RANK) expression at both gene and protein levels, linking it to a decrease in OC differentiation. An endogenous miR-218-2 block rescued the expression of RANK and subsequent OC formation in LPS-stimulated OCPs. Aligned with these results, SLIT3-deficient mice displayed increased OC formation and reduced bone density after LPS challenge. CONCLUSION: Our findings suggest that the TLR4-dependent concomitant induction of Slit3 and miR-218-2 targets RANK in OCPs to restrain OC commitment, thereby avoiding an uncoordinated loss of bone through inflammatory processes. These observations provide a mechanistic explanation for the role of TLR4 in controlling the commitment phase of OC differentiation. Video Abstract.

Asunto(s)

Osteoclastos , Receptor Toll-Like 4 , Animales , Ratones , Proteína beta Potenciadora de Unión a CCAAT , Lipopolisacáridos/farmacología , Macrófagos , Proteínas de la Membrana/genética , Proteínas del Tejido Nervioso/genética , Receptores Inmunológicos/genética

RESUMEN

Dendritic cell immunoreceptor (DCIR) is an inhibitory C-type lectin receptor that acts as a negative regulator in the immune system and bone metabolism. We previously revealed that DCIR deficiency enhanced osteoclastogenesis and antigen presentation of dendritic cells, and that asialo-biantennary N-glycan (NA2) functions as a ligand for DCIR. NA2 binding to DCIR suppressed murine and human osteoclastogenesis that occurs in the presence of M-CSF and RANKL. The DCIR-NA2 axis, therefore, plays an important role in regulating osteoclastogenesis in both mice and humans, although the underlying mechanisms remain unclear. Here we found that Dcir -/- bone marrow-derived macrophages (BMMs) exhibited greater proliferative and differentiation responses to M-CSF and RANKL, respectively, than wild-type (WT) BMMs. Moreover, Dcir -/- osteoclasts (OCs) increased resorptive activity and cell fusion more significantly than WT OCs. DCIR deficiency affects gene expression patterns in OCs, and we found that the expression of neuraminidase 4 was increased in Dcir -/- OCs. Furthermore, DCIR-NA2 interaction in WT BMMs, but not Dcir -/- BMMs, decreased Akt phosphorylation in response to M-CSF and RANKL. These data suggest that DCIR regulates osteoclastogenesis by downregulating M-CSF and RANKL signaling, and that DCIR-mediated signaling may contribute to the terminal modification of oligosaccharides by controlling the expression of glycosylation enzymes.

Asunto(s)

Resorción Ósea , Factor Estimulante de Colonias de Macrófagos , Animales , Humanos , Ratones , Resorción Ósea/metabolismo , Proliferación Celular , Factor Estimulante de Colonias de Macrófagos/farmacología , Factor Estimulante de Colonias de Macrófagos/metabolismo , Osteoclastos/metabolismo , Transducción de Señal

RESUMEN

Dendritic cell immunoreceptor (DCIR; Clec4a2), a member of the C-type lectin receptor family, plays important roles in homeostasis of the immune and bone systems. However, the intestinal role of this molecule is unclear. Here, we show that dextran sodium sulfate (DSS)-induced colitis and azoxymethane-DSS-induced intestinal tumors are reduced in Clec4a2-/- mice independently of intestinal microbiota. STAT5 phosphorylation and expression of Csf2 and tight junction genes are enhanced, while Il17a and Cxcl2 are suppressed in the Clec4a2-/- mouse colon, which exhibits reduced infiltration of neutrophils and myeloid-derived suppressor cells. Granulocyte-macrophage colony-stimulating factor (GM-CSF) administration ameliorates DSS colitis associated with reduced Il17a and enhanced tight junction gene expression, whereas anti-GM-CSF exacerbates symptoms. Furthermore, anti-NA2, a ligand for DCIR, ameliorates colitis and prevents colorectal tumors. These observations indicate that blocking DCIR signaling ameliorates colitis and suppresses colonic tumors, suggesting DCIR as a possible target for the treatment of these diseases.

Asunto(s)

Colitis , Neoplasias Colorrectales , Animales , Colitis/patología , Sulfato de Dextran , Modelos Animales de Enfermedad , Factor Estimulante de Colonias de Granulocitos y Macrófagos/metabolismo , Lectinas Tipo C/genética , Lectinas Tipo C/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Factor de Transcripción STAT5/metabolismo

RESUMEN

Clec1A, a member of C-type lectin receptor family, has a carbohydrate recognition domain in its extracellular region, but no known signaling motif in the cytoplasmic domain. Clec1a is highly expressed in endothelial cells and weakly in dendritic cells. Although this molecule was reported to play an important role in the host defense against Aspergillus fumigatus by recognizing 1,8-dihydroxynaphthalene-melanin on the fungal surface, the roles of this molecule in un-infected animals remain to be elucidated. In this study, we found that Clec1a-/- mice develop milder symptoms upon induction of experimental autoimmune encephalomyelitis (EAE), an animal model for multiple sclerosis. The maximum disease score was significantly lower, and demyelination and inflammation of the spinal cord were much milder in Clec1a-/- mice compared to wild-type mice. No abnormality was detected in the immune cell composition in the draining lymph nodes and spleen on day 10 and 16 after EAE induction. Recall memory T cell proliferation after restimulation with myelin oligodendrocyte glycoprotein peptide (MOG35-55) in vitro was decreased in Clec1a-/- mice, and antigen presenting ability of Clec1a-/- dendritic cells was impaired. Interestingly, RNA-Seq and RT-qPCR analyses clearly showed that the expression of inflammatory cytokines including Il17a, Il6 and Il1b was greatly decreased in Clec1a-/- mice after induction of EAE, suggesting that this reduced cytokine production is responsible for the amelioration of EAE in Clec1a-/- mice. These observations suggest a novel function of Clec1A in the immune system.

Asunto(s)

Presentación de Antígeno , Células Dendríticas , Encefalomielitis Autoinmune Experimental , Interleucina-17 , Lectinas Tipo C , Animales , Citocinas/metabolismo , Células Dendríticas/metabolismo , Encefalomielitis Autoinmune Experimental/metabolismo , Encefalomielitis Autoinmune Experimental/patología , Células Endoteliales/metabolismo , Interleucina-17/metabolismo , Lectinas Tipo C/genética , Lectinas Tipo C/metabolismo , Ratones , Ratones Endogámicos C57BL

RESUMEN

Dendritic cell immunoreceptor (DCIR) is a C-type lectin receptor with a carbohydrate recognition domain and an immunoreceptor tyrosine-based inhibitory motif. Previously, we showed that Dcir-/- mice spontaneously develop autoimmune enthesitis and sialadenitis, and also develop metabolic bone abnormalities. However, the ligands for DCIR functionality remain to be elucidated. Here we showed that DCIR is expressed on osteoclasts and DCs and binds to an asialo-biantennary N-glycan(s) (NA2) on bone cells and myeloid cells. Osteoclastogenesis was enhanced in Dcir-/- cells, and NA2 inhibited osteoclastogenesis. Neuraminidase treatment, which exposes excess NA2 by removing the terminal sialic acid of N-glycans, suppressed osteoclastogenesis and DC function. Neuraminidase treatment of mice ameliorated collagen-induced arthritis and experimental autoimmune encephalomyelitis in a DCIR-dependent manner, due to suppression of antigen presentation by DCs. These results suggest that DCIR activity is regulated by the modification of the terminal sialylation of biantennary N-glycans, and this interaction is important for the control of both autoimmune and bone metabolic diseases.

Asunto(s)

Células Dendríticas/fisiología , Lectinas Tipo C/metabolismo , Glicoproteínas de Membrana/metabolismo , Osteoclastos/metabolismo , Osteogénesis/fisiología , Polisacáridos/metabolismo , Receptores Inmunológicos/metabolismo , Animales , Artritis Experimental/inducido químicamente , Artritis Experimental/tratamiento farmacológico , Células Cultivadas , Células Dendríticas/inmunología , Encefalomielitis Autoinmune Experimental/tratamiento farmacológico , Células HEK293 , Humanos , Lectinas Tipo C/genética , Glicoproteínas de Membrana/genética , Ratones Endogámicos C57BL , Ratones Mutantes , Ratones Transgénicos , N-Acetilglucosaminiltransferasas/genética , N-Acetilglucosaminiltransferasas/metabolismo , Neuraminidasa/metabolismo , Neuraminidasa/farmacología , Receptores Inmunológicos/genética

RESUMEN

The interleukin-17 (IL-17) family consists of six family members (IL-17A-IL-17F) and all the corresponding receptors have been identified recently. This family is mainly involved in the host defense mechanisms against bacteria, fungi and helminth infection by inducing cytokines and chemokines, recruiting neutrophils, inducing anti-microbial proteins and modifying T-helper cell differentiation. IL-17A and some other family cytokines are also involved in the development of psoriasis, psoriatic arthritis and ankylosing spondylitis by inducing inflammatory cytokines and chemokines, and antibodies against IL-17A as well as the receptor IL-17RA are being successfully used for the treatment of these diseases. Involvement in the development of inflammatory bowel disease, multiple sclerosis, rheumatoid arthritis and tumors has also been suggested in animal disease models. In this review, we will briefly review the mechanisms by which IL-17 cytokines are involved in the development of these diseases and discuss possible treatment of inflammatory diseases by targeting IL-17 family members.

Asunto(s)

Artritis Reumatoide/inmunología , Enfermedades Inflamatorias del Intestino/inmunología , Interleucina-17/inmunología , Esclerosis Múltiple/inmunología , Neoplasias/inmunología , Animales , Humanos

RESUMEN

Aflatoxins (AFs) are biologically active toxic metabolites, which are produced by certain toxigenic Aspergillus sp. on agricultural crops. In this study, five edible mushroom-forming fungi were analyzed using high-performance liquid chromatography fluorescence detector (HPLC-FLD) for their ability to remove aflatoxin B1 (AFB1), one of the most potent naturally occurring carcinogens known. Bjerkandera adusta and Auricularia auricular-judae showed the most significant AFB1 removal activities (96.3% and 100%, respectively) among five strains after 14-day incubation. The cell lysate from B. adusta exhibited higher AFB1 removal activity (35%) than the cell-free supernatant (13%) after 1-day incubation and the highest removal activity (80%) after 5-day incubation at 40 °C. In addition, AFB1 analyses using whole cells, cell lysates, and cell debris from B. adusta showed that cell debris had the highest AFB1 removal activity at 5th day (95%). Moreover, exopolysaccharides from B. adusta showed an increasing trend (24-48%) similar to whole cells and cell lysates after 5- day incubation. Our results strongly suggest that AFB1 removal activity by whole cells was mainly due to AFB1 binding onto cell debris during early incubation and partly due to binding onto cell lysates along with exopolysaccharides after saturation of AFB1 binding process onto cell wall components.

Asunto(s)

Aflatoxina B1/metabolismo , Aflatoxina B1/toxicidad , Agaricales/metabolismo , Aspergillus/química , Aspergillus/metabolismo , Biodegradación Ambiental , Contaminación de Alimentos , Auricularia/metabolismo , Coriolaceae/metabolismo , Productos Agrícolas/microbiología , Hericium/metabolismo , República de Corea , Hongos Shiitake/metabolismo , Wolfiporia/metabolismo

RESUMEN

Understanding the host anti-fungal immunity induced by beta-glucan has been one of the most challenging conundrums in the field of biomedical research. During the last couple of decades, insights on the role of beta-glucan in fungal disease progression, susceptibility, and resistance have been greatly augmented through the utility of various beta-glucan cognate receptor-deficient mouse models. Analysis of dectin-1 knockout mice has clarified the downstream signaling pathways and adaptive effector responses triggered by beta-glucan in anti-fungal immunity. On the other hand, assessment of CR3-deficient mice has elucidated the compelling action of beta-glucans in neutrophil-mediated fungal clearance, and the investigation of EphA2-deficient mice has highlighted its novel involvement in host sensing and defense to oral mucosal fungal infection. Based on these accounts, this review focuses on the recent discoveries made by these gene-targeted mice in beta-glucan research with particular emphasis on the multifaceted aspects of fungal immunity.

Asunto(s)

Hongos/inmunología , Micosis/inmunología , beta-Glucanos/inmunología , Inmunidad Adaptativa , Animales , Modelos Animales de Enfermedad , Eliminación de Gen , Humanos , Inmunidad , Lectinas Tipo C/genética , Lectinas Tipo C/inmunología , Antígeno de Macrófago-1/genética , Antígeno de Macrófago-1/inmunología , Ratones , Ratones Noqueados , Micosis/genética , Micosis/microbiología , Receptor EphA2/genética , Receptor EphA2/inmunología

RESUMEN

TARM1 is a member of the leukocyte immunoglobulin-like receptor family and stimulates macrophages and neutrophils in vitro by associating with FcRγ. However, the function of this molecule in the regulation of the immune system is unclear. Here, we show that Tarm1 expression is elevated in the joints of rheumatoid arthritis mouse models, and the development of collagen-induced arthritis (CIA) is suppressed in Tarm1-/- mice. T cell priming against type 2 collagen is suppressed in Tarm1-/- mice and antigen-presenting ability of GM-CSF-induced dendritic cells (GM-DCs) from Tarm1-/- mouse bone marrow cells is impaired. We show that type 2 collagen is a functional ligand for TARM1 on GM-DCs and promotes DC maturation. Furthermore, soluble TARM1-Fc and TARM1-Flag inhibit DC maturation and administration of TARM1-Fc blocks the progression of CIA in mice. These results indicate that TARM1 is an important stimulating factor of dendritic cell maturation and could be a good target for the treatment of autoimmune diseases.

Asunto(s)

Artritis Experimental/metabolismo , Artritis Experimental/patología , Colágeno/metabolismo , Células Dendríticas/patología , Receptores Inmunológicos/metabolismo , Animales , Presentación de Antígeno , Factor Estimulante de Colonias de Granulocitos y Macrófagos/farmacología , Proteínas Fluorescentes Verdes/metabolismo , Inmunización , Ligandos , Ratones Endogámicos C57BL , Receptores Inmunológicos/deficiencia

RESUMEN

This study investigated the distribution of twelve mycotoxins (aflatoxins B1, B2, G1, and G2; ochratoxin A; fumonisins B1 and B2; deoxynivalenol; nivalenol; zearalenone; T-2 toxin; and HT-2 toxin) in corn and corn by-products (corn bran, cornstarch, corn gluten, corn gluten feed, corn germ, light steep water, and corn steep liquor) produced by wet-milling in Korea. Fifty-two samples were collected from three factories producing cornstarch and other corn by-products. The samples were pretreated on an immunoaffinity column (IAC), and then the levels of the 12 mycotoxins were analyzed simultaneously by liquid chromatography-coupled triple-quadrupole mass spectrometry (LC-MS/MS). Fusarium mycotoxins were mainly found in raw corn and corn gluten feed samples. Other mycotoxins-such as aflatoxins, ochratoxin A, and HT-2 toxin-were detected in tiny amounts below the limit of quantification (LOQ) in cornstarch, corn germ, and corn bran. Ochratoxin A and nivalenol were mainly carried over into cornstarch. Aflatoxin B1, deoxynivalenol, T-2 toxin, HT-2 toxin, and the fumonisins were concentrated in corn gluten feed. Zearalenone was evenly distributed in all corn by-products except cornstarch during the milling process.

Asunto(s)

Contaminación de Alimentos/análisis , Micotoxinas/análisis , Zea mays/química , Cromatografía Liquida , Grano Comestible/química , Manipulación de Alimentos , República de Corea , Espectrometría de Masas en Tándem

RESUMEN

IL-36α (gene symbol Il1f6), a member of the IL-36 family, is closely associated with inflammatory diseases, including colitis and psoriasis. In this study, we found that Il1f6-/- mice developed milder psoriasiform dermatitis upon treatment with imiquimod, a ligand for TLR ligand 7 (TLR7) and TLR8, whereas Il1f6-/- mice showed similar susceptibility to dextran sodium sulfate-induced colitis to wild-type mice. These effects were observed in both cohoused and separately housed conditions, and antibiotic treatment did not cancel the resistance of Il1f6-/- mice to imiquimod-induced dermatitis. Bone marrow (BM) cell transfer revealed that IL-36α expression in skin-resident cells is important for the pathogenesis of dermatitis in these mice. Following stimulation with IL-36α, the expression of Il1f6 and Il1f9 (IL-36γ), but not Il1f8 (IL-36ß), was enhanced in murine BM-derived Langerhans cells (BMLCs) and murine primary keratinocytes but not in fibroblasts from mice. Upon stimulation with agonistic ligands of TLRs and C-type lectin receptors (CLRs), Il1f6 expression was induced in BMLCs and BM-derived dendritic cells. Furthermore, IL-36α stimulation resulted in significantly increased gene expression of psoriasis-associated Th17-related cytokines and chemokines such as IL-1α, IL-1ß, IL-23, CXCL1, and CXCL2 in BMLCs and fibroblasts, and IL-1α, IL-1ß, IL-17C, and CXCL2 in keratinocytes. Collectively, these results suggest that TLR/CLR signaling-induced IL-36α plays an important role for the development of psoriasiform dermatitis by enhancing Th17-related cytokine/chemokine production in skin-resident cells via a local autoamplification loop.

Asunto(s)

Adyuvantes Inmunológicos/toxicidad , Quimiocinas/biosíntesis , Colitis/patología , Imiquimod/toxicidad , Interleucina-1/metabolismo , Queratinocitos/metabolismo , Psoriasis/patología , Piel/patología , Células Th17/inmunología , Animales , Células de la Médula Ósea/citología , Trasplante de Médula Ósea , Células Cultivadas , Colitis/inducido químicamente , Células Dendríticas/metabolismo , Sulfato de Dextran/toxicidad , Fibroblastos/metabolismo , Interleucina-1/genética , Células de Langerhans/metabolismo , Glicoproteínas de Membrana/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Psoriasis/tratamiento farmacológico , Psoriasis/genética , Piel/citología , Receptor Toll-Like 7/metabolismo , Receptor Toll-Like 8/metabolismo

RESUMEN

The Fusarium fungi produce toxic substances called mycotoxins, which can cause disease and harmful effects in grains, livestock, and humans. Deoxynivalenol (DON), also known as vomitoxin, is one of the Fusarium mycotoxins that is known to cause vomiting in livestock. This study shows the occurrence of deoxynivalenol in feedstuffs (compound feed and feed ingredients) between 2009 and 2016 in South Korea. A total of 653 domestic samples were collected at five time points, including 494 compound feed samples and 159 feed ingredient samples. DON contamination levels were analyzed using high-performance liquid chromatography (HPLC) with pretreatment using an immunoaffinity column (IAC). The limit of detection (LOD) and the limit of quantification (LOQ) were estimated at 1-10 µg/kg and 3-35 µg/kg, respectively. Two compound feeds (two gestating sow feed samples) out of 160 pig feed samples exceeded the European Commission (EC) guidance value, while no feed ingredient samples exceeded the EC or South Korean guidance values. There were statistically significant differences in the mean contamination levels of compound feed and feed ingredients that indicated a decreasing trend over time.

Asunto(s)

Alimentación Animal/análisis , Contaminación de Alimentos/análisis , Tricotecenos/análisis , Cromatografía Líquida de Alta Presión , Monitoreo del Ambiente , Límite de Detección , República de Corea

RESUMEN

Several plant pathogen Fusarium species produce fumonisins (FUMs); which can end up in food and feed and; when ingested; can exhibit harmful effects on humans and livestock. Mycotoxin intoxication by fumonisin B1 (FB1) and fumonisin B2 (FB2) can cause porcine pulmonary edema; leukoencephalomalacia in equines; esophageal cancer and birth defects by natural contamination. Herein; the occurrence of FB1 and FB2 in feedstuff (compound feed and feed ingredients) was investigated between 2011 and 2016 in South Korea. A total of 535 animal feed samples (425 compound feed samples and 110 feed ingredients) produced domestically were sampled four times between 2011 and 2016 (2011; 2012; 2014 and 2016) from feed factories in South Korea. The limit of detection (LOD) for FB1 and FB2 was 20 µg/kg and 25 µg/kg; respectively; and the limit of quantitation (LOQ) was 30 µg/kg and 35 µg/kg; respectively. The recovery range (%) was between 86.4% and 108.8%; and the relative standard deviation (RSD) (%) was 4.7-12.1%. Seven (swine feed samples) out of the 425 feed samples exceeded the European Union (EU) and South Korea commission regulations over the six-year test period; and no feed ingredients exceeded the guidelines.

Asunto(s)

Carcinógenos Ambientales/toxicidad , Contaminación de Alimentos , Fumonisinas/toxicidad , Alimentación Animal , Animales , Caballos , Humanos , República de Corea , Porcinos

RESUMEN

Mycotoxins produced by Fusarium plant pathogen species have harmful effects on humans and livestock by natural contamination in food and feed. Zearalenone, one of the well-known Fusarium mycotoxins, causes hyperestrogenism and toxicosis resulting in reproductive dysfunction in animals. This study investigated the occurrence of zearalenone in feedstuffs (compound feeds, feed ingredients) between 2009 and 2016 in South Korea to obtain information on zearalenone contamination in feeds for management. A total of 653 animal feed samples (494 compound feeds, 159 feed ingredients) produced domestically were sampled five times from 2009 to 2016 (2009, 2010, 2012, 2014, and 2016) from feed factories in South Korea. The levels of zearalenone were analyzed every year by high-performance liquid chromatography (HPLC) after pretreatment with an immunoaffinity column showing limit of detection (LOD) and limit of quantification (LOQ) of 0.1-3 µg/kg and 0.3-8 µg/kg, respectively. Four feed samples out of 494 compound feeds exceeded the EU and South Korea commission regulations over the eight-year test period, and no feed ingredients exceeded the guidelines.

Asunto(s)

Alimentación Animal/análisis , Contaminación de Alimentos/análisis , Zearalenona/análisis , Cromatografía Líquida de Alta Presión , Monitoreo del Ambiente/estadística & datos numéricos , República de Corea

RESUMEN

Interleukin-17 (IL-17)-producing γδ T (γδ17) cells have been implicated in inflammatory diseases, but the underlying pathogenic mechanisms remain unclear. Here, we show that both CD4(+) and γδ17 cells are required for the development of autoimmune arthritis in IL-1 receptor antagonist (IL-1Ra)-deficient mice. Specifically, activated CD4(+) T cells direct γδ T-cell infiltration by inducing CCL2 expression in joints. Furthermore, IL-17 reporter mice reveal that the Vγ6(+) subset of CCR2(+) γδ T cells preferentially produces IL-17 in inflamed joints. Importantly, because IL-1Ra normally suppresses IL-1R expression on γδ T cells, IL-1Ra-deficient mice exhibit elevated IL-1R expression on Vγ6(+) cells, which play a critical role in inducing them to produce IL-17. Our findings demonstrate a pathogenic mechanism in which adaptive and innate immunity induce an autoimmune disease in a coordinated manner.

Asunto(s)

Artritis/inmunología , Enfermedades Autoinmunes/metabolismo , Proteína Antagonista del Receptor de Interleucina 1/metabolismo , Interleucina-17/metabolismo , Activación de Linfocitos/fisiología , Subgrupos de Linfocitos T/fisiología , Animales , Artritis/metabolismo , Quimiocina CCL2/genética , Quimiocina CCL2/metabolismo , Regulación de la Expresión Génica/inmunología , Proteína Antagonista del Receptor de Interleucina 1/genética , Interleucina-17/genética , Articulaciones/metabolismo , Articulaciones/patología , Ratones , Ratones Noqueados , Receptores de Antígenos de Linfocitos T gamma-delta , Receptores Tipo I de Interleucina-1/genética , Receptores Tipo I de Interleucina-1/metabolismo , Subgrupos de Linfocitos T/inmunología

RESUMEN

Dendritic cell immunoreceptor (DCIR) is a C-type lectin receptor mainly expressed in DCs. Dcir (-/-) mice spontaneously develop autoimmune enthesitis and ankylosis accompanied by fibrocartilage proliferation and ectopic ossification. However, the mechanisms of new bone/cartilage formation in Dcir (-/-) mice remain to be elucidated. In this study, we show that DCIR maintains bone homeostasis by regulating IFN-γ production under pathophysiological conditions. DCIR deficiency increased bone volume in femurs and caused aberrant ossification in joints, whereas these symptoms were abolished in Rag2(-/-)Dcir(-/-) mice. IFN-γ-producing T cells accumulated in lymph nodes and joints of Dcir(-/-) mice, and purified Dcir(-/-) DCs enhanced IFN-γ(+) T cell differentiation. The ankylotic changes and bone volume increase were suppressed in the absence of IFN-γ. Thus, IFN-γ is a positive chondrogenic and osteoblastogenic factor, and DCIR is a crucial regulator of bone metabolism; consequently, both factors are potential targets for therapies directed against bone metabolic diseases.

Asunto(s)

Huesos/metabolismo , Homeostasis/genética , Homeostasis/inmunología , Interferón gamma/biosíntesis , Lectinas Tipo C/genética , Subgrupos de Linfocitos T/inmunología , Subgrupos de Linfocitos T/metabolismo , Factores de Edad , Animales , Anquilosis/diagnóstico por imagen , Anquilosis/genética , Anquilosis/inmunología , Anquilosis/patología , Densidad Ósea/genética , Huesos/diagnóstico por imagen , Huesos/patología , Diferenciación Celular/genética , Diferenciación Celular/inmunología , Condrocitos/citología , Condrocitos/metabolismo , Proteínas de Unión al ADN/deficiencia , Células Dendríticas/inmunología , Células Dendríticas/metabolismo , Masculino , Ratones , Ratones Noqueados , Osteoblastos/citología , Osteoblastos/metabolismo , Subgrupos de Linfocitos T/citología , Microtomografía por Rayos X