RESUMEN
Pregnant women are exposed to various microbes, some of which can harm the mother and/or fetus and can lead to life-long morbidity and even death. The syncytiotrophoblast (STB) covers the placental villi and comes into direct contact with pathogens contained in the maternal blood and plays a key role in placental host defense. However, the precise mechanisms whereby the STB recognizes and responds to pathogenic microbes remain unclear. In this study, we comprehensively analyzed the expression of functional pattern recognition receptors, which are responsible for tissue defense against pathogens, in a primary STB model differentiated from highly purified human term cytotrophoblasts (CTBs). Screening for mRNA expression and multiplex cytokine/chemokine production demonstrated that differentiated CTBs (dCTBs) predominantly expressed dsRNA receptors, including TLR3, MDA5, and RIG-I. We confirmed that term human placentas also expressed TLR3. Transcriptome analysis revealed common and unique responses of dCTBs to a synthetic dsRNA (polyinosinic-polycytidylic acid) compared with human peripheral mononuclear cells. Moreover, polyinosinic-polycytidylic acid induced the release of type I and type III IFNs (IFN-ß, IFN-λ1, IFN-λ2, IFN-λ3), as well as mRNA expression of IFN-stimulated genes (IFIT1, MX1, and OAS1). dCTBs underwent apoptosis via the mitochondrial pathway in response to dsRNA stimulation. These results suggest that dsRNA receptors expressed on the STB are key players in antiviral defense in the placenta. Elucidation of the underpinnings of these defense processes can help us better understand the pathophysiology of viral infections during pregnancy.
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Placenta , Trofoblastos , Humanos , Femenino , Embarazo , Placenta/metabolismo , Poli I-C/farmacología , Receptor Toll-Like 3/metabolismo , Receptores de Reconocimiento de Patrones/genética , ARN Bicatenario , ARN MensajeroRESUMEN
This study aimed to (1) determine whether the expression of angiotensin-converting enzyme 2 (ACE2) and transmembrane protease serine 2 is increased in tobacco smokers, which potentially increases their susceptibility to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, and (2) assess whether eye rinsing can reduce susceptibility. This prospective study included 20 eyes of 10 smokers and 18 eyes of nine healthy non-smokers (control) for reverse-transcription polymerase chain reaction. This study also included 28 eyes of 14 smokers and 16 eyes of eight healthy non-smokers (control) for enzyme-linked immunosorbent assay. Tear and impression cytology samples were collected from the right eye of each patient. The left eye was then rinsed for 30 s, and after 5 min, the tear and impression cytology samples were collected in the same manner. The expression of the ACE2 gene was significantly higher in the conjunctiva of smokers (n = 17; median 3.07 copies/ng of total RNA) than in those of non-smokers (n = 17; median 1.92 copies/ng of total RNA, p = 0.003). Further, mRNA expression and protein levels of ACE2 were weakly correlated in smokers (r = 0.49). ACE2 protein levels in Schirmer's strip samples were significantly reduced from 5051 to 3202 pg/mL after eye washing (n = 10; p = 0.001). Ocular surface cells are susceptible to SARS-CoV-2 infection. Smoking may be a risk factor for SARS-CoV-2 infection, and eye rinsing may reduce the risk of infection.
Asunto(s)
Enzima Convertidora de Angiotensina 2 , COVID-19 , Humanos , Enzima Convertidora de Angiotensina 2/genética , Enzima Convertidora de Angiotensina 2/metabolismo , Conjuntiva/metabolismo , COVID-19/metabolismo , COVID-19/prevención & control , Estudios Prospectivos , ARN/metabolismo , SARS-CoV-2/metabolismo , Fumadores , Ojo/metabolismoRESUMEN
BACKGROUND: Atopic keratoconjunctivitis (AKC) is a chronic allergic conjunctival disease. However, a mouse model of AKC to investigate the underlying mechanism of the therapeutic agents and estimate their efficacy has not been established. We recently generated mice in which Ikk2 is specifically deleted in facial skin fibroblasts and found that these mice spontaneously develop atopic dermatitis (AD)-like facial skin inflammation and scratching behaviors; thus, we named them facial AD with scratching (FADS) mice. OBJECTIVE: We sought to evaluate whether the ocular lesions that FADS mice spontaneously develop are similar to those of patients with AKC and to estimate the efficacy of topical treatments with tacrolimus and betamethasone for FADS mice by using tear periostin, a novel biomarker for allergic conjunctival disease. METHODS: FADS mice, in which Ikk2 is deleted in dermal fibroblasts, were generated by crossing female Ikk2Flox/Flox mice to male Nestincre; Ikk2Flox/+ mice. We conducted histologic analysis of the ocular lesions in FADS mice. Furthermore, we measured periostin in the tears collected from FADS mice untreated or treated with tacrolimus or betamethasone. RESULTS: The FADS mice exhibited severe blepharitis and scratch behaviors for their faces. In these mice, corneal epithelium and stroma showed hyperplasia and infiltration of eosinophils, mast cells, and TH2/TC2 cells. Periostin was significantly expressed in the lesions and tear periostin was upregulated. Betamethasone showed more suppressive effects than did tacrolimus on severe corneal lesions and increased tear periostin level. CONCLUSIONS: The FADS mouse is a novel mouse model of AKC and is useful to examine the therapeutic effects of anti-AKC agents.
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Blefaritis/genética , Fibroblastos/fisiología , Hipersensibilidad Inmediata/genética , Quinasa I-kappa B/genética , Queratoconjuntivitis/genética , Nestina/genética , Piel/patología , Animales , Blefaritis/inmunología , Moléculas de Adhesión Celular/metabolismo , Modelos Animales de Enfermedad , Humanos , Hipersensibilidad Inmediata/inmunología , Inmunidad Celular , Queratoconjuntivitis/inmunología , Ratones , Ratones Noqueados , Lágrimas/metabolismoRESUMEN
BACKGROUND: Group 2 innate lymphoid cells (ILC2s) play critical roles in induction and exacerbation of allergic airway inflammation. Thus clarification of the mechanisms that underlie regulation of ILC2 activation has received significant attention. Although innate lymphoid cells are divided into 3 major subsets that mirror helper effector T-cell subsets, counterpart subsets of regulatory T cells have not been well characterized. OBJECTIVE: We sought to determine the factors that induce regulatory innate lymphoid cells (ILCregs). METHODS: IL-10+ ILCregs induced from ILC2s by using retinoic acid (RA) were analyzed with RNA-sequencing and flow cytometry. ILCregs were evaluated in human nasal tissue from healthy subjects and patients with chronic rhinosinusitis with nasal polyps and lung tissue from house dust mite- or saline-treated mice. RESULTS: RA induced IL-10 secretion by human ILC2s but not type 2 cytokines. IL-10+ ILCregs, which were converted from ILC2s by means of RA stimulation, expressed a regulatory T cell-like signature with expression of IL-10, cytotoxic T lymphocyte-associated protein 4, and CD25, with downregulated effector type 2-related markers, such as chemoattractant receptor-homologous molecule on TH2 cells and ST2, and suppressed activation of CD4+ T cells and ILC2s. ILCregs were rarely detected in human nasal tissue from healthy subjects or lung tissue from saline-treated mice, but numbers were increased in nasal tissue from patients with chronic rhinosinusitis with nasal polyps and in lung tissue from house dust mite-treated mice. Enzymes for RA synthesis were upregulated in airway epithelial cells during type 2 inflammation in vivo and by IL-13 in vitro. CONCLUSION: We have identified a unique immune regulatory and anti-inflammatory pathway by which RA converts ILC2s to ILCregs. Interactions between airway epithelial cells and ILC2s play an important roles in the generation of ILCregs.
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Antiinflamatorios/farmacología , Linfocitos/efectos de los fármacos , Tretinoina/farmacología , Animales , Línea Celular , Citocinas/inmunología , Células Epiteliales/inmunología , Humanos , Inmunidad Innata , Pulmón/inmunología , Linfocitos/inmunología , Ratones Endogámicos C57BL , Senos Paranasales/inmunologíaRESUMEN
BACKGROUND: Severe atopic keratoconjunctivitis (AKC) is a relatively rare disease, and some cases are refractory to conventional steroid treatment. OBJECTIVE: To examine the efficacy of 0.1% tacrolimus ophthalmic suspension in treating severe AKC during a 1-year follow-up. METHODS: This was a single-center, retrospective clinical study. Sixty eyes from 30 patients with severe AKC who were treated with 0.1% tacrolimus ophthalmic suspension 4 times per day, were included. The mean age of the patients was 21.5 ± 13.7 years. The severity of objective signs was observed at baseline (before treatment), at 2 weeks, and at 1, 2, 3, 6, and 12 months after treatment initiation. Ten objective signs of palpebral conjunctiva, bulbar conjunctiva, limbus, and cornea were assessed using 4 grades (0 = normal; 1+ = mild; 2+ = moderate; 3+ = severe). Safety was assessed based on the incidence and the severity of adverse events. RESULTS: The total score of the 10 clinical signs significantly decreased from baseline 2 weeks after initiating tacrolimus eye drop treatment, except at 2 months. The mean total score of clinical signs was 13.6 ± 6.6 at the beginning of treatment, and decreased to 5.4 ± 4.8 12 months after initiation. Treatment was gradually tapered, with increasing intervals between applications. Additional medications were required to provide relief in 18 patients during follow-up. No patient discontinued treatment due to adverse drug effects. Herpes keratitis was observed in 3 cases during follow-up. However, these cases were completely controlled. CONCLUSION: The 0.1% tacrolimus ophthalmic suspension is effective for the treatment of severe AKC refractory to standard conventional treatments throughout a full year.
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Conjuntivitis Alérgica/tratamiento farmacológico , Inmunosupresores/administración & dosificación , Queratoconjuntivitis/tratamiento farmacológico , Tacrolimus/administración & dosificación , Adolescente , Adulto , Niño , Femenino , Humanos , Inmunosupresores/efectos adversos , Masculino , Soluciones Oftálmicas , Estudios Retrospectivos , Índice de Severidad de la Enfermedad , Tacrolimus/efectos adversos , Resultado del Tratamiento , Adulto JovenRESUMEN
BACKGROUND: Migration of cancer cell correlates with distant metastasis and local invasion, which are good targets for cancer treatment. An optically accessible device "TAXIScan" was developed, which provides considerably more information regarding the cellular dynamics and less quantity of samples than do the existing methods. Here, we report the establishment of a system to analyze the nature of pancreatic cancer cells using TAXIScan and we evaluated lysophosphatidic acid (LPA)-elicited pancreatic cell migration. METHODS: Pancreatic cancer cell lines, BxPC3, PANC-1, AsPC1, and MIAPaCa-2, were analyzed for adhesion as well as migration towards LPA by TAXIScan using parameters such as velocity and directionality or for the number of migrated cells by the Boyden chamber methods. To confirm that the migration was initiated by LPA, the expression of LPA receptors and activation of intracellular signal transductions were examined by quantitative reverse transcriptase polymerase reaction and western blotting. RESULTS: Scaffold coating was necessary for the adhesion of pancreatic cancer cells, and collagen I and Matrigel were found to be good scaffolds. BxPC3 and PANC-1 cells clearly migrated towards the concentration gradient formed by injecting 1 µL LPA, which was abrogated by pre-treatment with LPA inhibitor, Ki16425 (IC50 for the directionality ≈ 1.86 µM). The LPA dependent migration was further confirmed by mRNA and protein expression of LPA receptors as well as phosphorylation of signaling molecules. LPA1 mRNA was highest among the 6 receptors, and LPA1, LPA2 and LPA3 proteins were detected in BxPC3 and PANC-1 cells. Phosphorylation of Akt (Thr308 and Ser473) and p42/44MAPK in BxPC3 and PANC-1 cells was observed after LPA stimulation, which was clearly inhibited by pre-treatment with a compound Ki16425. CONCLUSIONS: We established a novel pancreatic cancer cell migration assay system using TAXIScan. This assay device provides multiple information on migrating cells simultaneously, such as their morphology, directionality, and velocity, with a small volume of sample and can be a powerful tool for analyzing the nature of cancer cells and for identifying new factors that affect cell functions.
Asunto(s)
Movimiento Celular/fisiología , Lisofosfolípidos/farmacología , Dispositivos Ópticos/estadística & datos numéricos , Neoplasias Pancreáticas/patología , Movimiento Celular/efectos de los fármacos , Humanos , Técnicas In Vitro , Neoplasias Pancreáticas/tratamiento farmacológico , Receptores del Ácido Lisofosfatídico/metabolismo , Transducción de Señal/efectos de los fármacos , Células Tumorales CultivadasRESUMEN
BACKGROUND: Chronic ocular allergic diseases such as vernal keratoconjunctivitis (VKC) and atopic keratoconjunctivitis (AKC) are accompanied by serious comorbidities; however, the underlying pathogenesis remains obscure. Furthermore, diagnosing conjunctival lesions in patients with atopic dermatitis and estimating the severity in AKC are important for the treatment of ocular allergic diseases. OBJECTIVE: We addressed whether periostin, a novel mediator and biomarker in allergic inflammation, is involved in the pathogenesis of ocular allergic diseases and whether periostin can be a biomarker for these diseases. METHODS: We investigated tear periostin in patients with seasonal allergic conjunctivitis (SAC), VKC, and AKC and allergic patients without conjunctivitis and compared it with tear IL-13 and serum periostin. Furthermore, in patients with AKC, we measured tear periostin before and after topical treatment with tacrolimus. RESULTS: Tears from patients with ocular allergic disease showed significantly high periostin levels than did tears from allergic patients without conjunctivitis and from patients with AKC, VKC, and SAC in descending order. Tear periostin was associated with serious comorbidities such as large papilla formation and corneal damage in AKC, although both tear IL-13 and serum periostin had little to no such abilities. Furthermore, after topical tacrolimus treatment, tear periostin tended to decrease in most patients with AKC along with their clinical improvement. CONCLUSIONS: Periostin produced in conjunctival tissues stimulated by IL-13 may contribute to the pathogenesis of ocular allergic diseases. Furthermore, tear periostin can be potentially applied as a biomarker to diagnose conjunctivitis in allergic patients and to evaluate disease severity as well as the efficacy of treatments in AKC.
Asunto(s)
Moléculas de Adhesión Celular/metabolismo , Oftalmopatías/diagnóstico , Oftalmopatías/metabolismo , Hipersensibilidad/diagnóstico , Hipersensibilidad/metabolismo , Lágrimas/metabolismo , Biomarcadores , Estudios de Casos y Controles , Enfermedad Crónica , Conjuntivitis Alérgica/diagnóstico , Conjuntivitis Alérgica/metabolismo , Manejo de la Enfermedad , Ensayo de Inmunoadsorción Enzimática , Células Epiteliales/metabolismo , Femenino , Fibroblastos/metabolismo , Humanos , Interleucina-13/sangre , Interleucina-13/metabolismo , MasculinoRESUMEN
BACKGROUND: Although platelets play a key role in allergic inflammation in addition to their well-established role in hemostasis, the precise mechanisms of how platelets modulate allergic inflammation are not fully understood. IL-33 is an essential regulator of innate immune responses and allergic inflammation. OBJECTIVE: We sought to determine the expression of IL-33 protein by platelets and its functional significance in airway inflammation. METHODS: IL-33 protein in human platelets, the human megakaryocyte cell line MEG-01, and bone marrow-derived mouse megakaryocytes was detected by using Western blot analysis and fluorescent immunostaining. We examined the functional relevance of IL-33 protein in platelets by comparing platelet-intact and platelet-depleted groups in a murine model of IL-33-dependent airway eosinophilia elicited by intranasal administration of papain. We further compared the additive effect of administration of platelets derived from wild-type versus IL-33-deficient mice on the papain-induced eosinophilia. RESULTS: Platelets and their progenitor cells, megakaryocytes, constitutively expressed IL-33 protein (31 kDa). Papain-induced IL-33-dependent airway eosinophilia in mice was significantly attenuated by platelet depletion. Conversely, concomitant administration of platelets derived from wild-type mice but not IL-33-deficient mice enhanced the papain-induced airway eosinophilia. CONCLUSIONS: Our novel findings suggest that platelets might be important cellular sources of IL-33 protein in vivo and that platelet-derived IL-33 might play a role in airway inflammation. Therefore platelets might become an attractive novel therapeutic target for asthma and probably allergic inflammation.
Asunto(s)
Plaquetas/inmunología , Citocinas/inmunología , Eosinofilia Pulmonar/inmunología , Animales , Líquido del Lavado Bronquioalveolar/citología , Líquido del Lavado Bronquioalveolar/inmunología , Células COS , Recuento de Células , Línea Celular , Citocinas/genética , Femenino , Humanos , Pulmón/inmunología , Ratones Endogámicos C57BL , Ratones Noqueados , Papaína , Eosinofilia Pulmonar/inducido químicamente , ARN Mensajero/metabolismoRESUMEN
BACKGROUND: Eosinophils play important roles in asthma, especially airway remodeling, by producing various granule proteins, chemical mediators, cytokines, chemokines and proteases. However, protease production by eosinophils is not fully understood. In the present study, we investigated the production of eosinophil-specific proteases/proteinases by transcriptome analysis. METHODS: Human eosinophils and other cells were purified from peripheral blood by density gradient sedimentation and negative/positive selections using immunomagnetic beads. Protease/proteinase expression in eosinophils and release into the supernatant were evaluated by microarray analysis, qPCR, ELISA, flow cytometry and immunofluorescence staining before and after stimulation with eosinophil-activating cytokines and secretagogues. mRNAs for extracellular matrix proteins in human normal fibroblasts were measured by qPCR after exposure to recombinant protease serine 33 (PRSS33) protein (rPRSS33), created with a baculovirus system. RESULTS: Human eosinophils expressed relatively high levels of mRNA for metalloproteinase 25 (MMP25), a disintegrin and metalloprotease 8 (ADAM8), ADAM10, ADAM19 and PRSS33. Expression of PRSS33 was the highest and eosinophil-specific. PRSS33 mRNA expression was not affected by eosinophil-activating cytokines. Immunofluorescence staining showed that PRSS33 was co-localized with an eosinophil granule protein. PRSS33 was not detected in the culture supernatant of eosinophils even after stimulation with secretagogues, but its cell surface expression was increased. rPRSS33 stimulation of human fibroblasts increased expression of collagen and fibronectin mRNAs, at least in part via protease-activated receptor-2 activation. CONCLUSIONS: Activated eosinophils may induce fibroblast extracellular matrix protein synthesis via cell surface expression of PRSS33, which would at least partly explain eosinophils' role(s) in airway remodeling.
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Eosinófilos/metabolismo , Expresión Génica , Serina Proteasas/genética , Serina Proteasas/metabolismo , Asma/inmunología , Asma/metabolismo , Asma/patología , Biomarcadores , Degranulación de la Célula/inmunología , Células Cultivadas , Eosinófilos/inmunología , Fibroblastos/metabolismo , Citometría de Flujo , Perfilación de la Expresión Génica , Humanos , Leucocitos/inmunología , Leucocitos/metabolismo , Especificidad de Órganos/genética , Transporte de Proteínas , TranscriptomaRESUMEN
ABSTRACT: A 76-year-old woman with liver and bone metastasis of a duodenal neuroendocrine tumor received peptide receptor radionuclide therapy with 177 Lu-DOTATATE. Scintigraphy with SPECT/CT performed 4 days after the treatment demonstrated 177 Lu-DOTATATE uptake as multifocal ground glass opacities in the bilateral lungs. This uptake was considered to be due to COVID-19 pneumonia because the patient was infected with the virus 7 days prior to the treatment. The lung opacities became smaller, showing a decreased uptake, 2 months later, after the second treatment. 177 Lu-DOTATATE may be taken up during the active phase of COVID-19 pneumonia.
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COVID-19 , Pulmón , Octreótido , Compuestos Organometálicos , Neumonía Viral , Humanos , COVID-19/diagnóstico por imagen , COVID-19/complicaciones , Anciano , Femenino , Pulmón/diagnóstico por imagen , Octreótido/análogos & derivados , Octreótido/uso terapéutico , Neumonía Viral/diagnóstico por imagen , Neumonía Viral/complicaciones , Pandemias , Tomografía Computarizada por Tomografía Computarizada de Emisión de Fotón Único , Infecciones por Coronavirus/diagnóstico por imagen , Infecciones por Coronavirus/complicaciones , Tumores Neuroendocrinos/diagnóstico por imagen , Tumores Neuroendocrinos/radioterapiaRESUMEN
Immune check point inhibitors (ICIs) have durable antitumor effects. However, autoimmune toxicities, termed immune-related adverse events, occur in some patients. We report a case of severe immune aplastic anemia (AA) in a patient with non-small cell lung cancer who was receiving atezolizumab with bevacizumab/carboplatin/paclitaxel. Although the cancer has not recurred, his bone marrow is depleted and he did not respond to immunosuppressive therapy. He has survived for 1.5 years with blood transfusions and infection control. Immune AA associated with ICIs is rare, and a treatment has not yet been established. This case report provides information on the management and treatment response of patients with AA caused by ICIs. Further studies should investigate the mechanism and pathogenesis of immune AA caused by ICIs.
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Anemia Aplásica , Anticuerpos Monoclonales Humanizados , Protocolos de Quimioterapia Combinada Antineoplásica , Bevacizumab , Carboplatino , Carcinoma de Pulmón de Células no Pequeñas , Neoplasias Pulmonares , Paclitaxel , Humanos , Carcinoma de Pulmón de Células no Pequeñas/tratamiento farmacológico , Anemia Aplásica/inducido químicamente , Neoplasias Pulmonares/tratamiento farmacológico , Anticuerpos Monoclonales Humanizados/efectos adversos , Anticuerpos Monoclonales Humanizados/administración & dosificación , Masculino , Protocolos de Quimioterapia Combinada Antineoplásica/efectos adversos , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Carboplatino/administración & dosificación , Carboplatino/efectos adversos , Bevacizumab/efectos adversos , Bevacizumab/administración & dosificación , Paclitaxel/administración & dosificación , Paclitaxel/efectos adversos , Inhibidores de Puntos de Control Inmunológico/efectos adversos , Inhibidores de Puntos de Control Inmunológico/administración & dosificación , Resultado del Tratamiento , Persona de Mediana Edad , Índice de Severidad de la EnfermedadRESUMEN
A 73-year-old man with lung squamous cell carcinoma was administered carboplatin + nab-paclitaxel + pembrolizumab for four cycles. Subsequently, he presented with multiple purpuras on his extremities, joint swelling on his fingers, abdominal pain, and diarrhea, accompanied by acute kidney injury (AKI), increased proteinuria, hematuria, and elevated C-reactive protein levels. Skin biopsy showed leukocytoclastic vasculitis as well as IgA and C3 deposition in the vessel walls. Based on these findings, the patient was diagnosed with IgA vasculitis as an immune-related adverse event (irAE) induced by carboplatin + nab-paclitaxel + pembrolizumab. After discontinuation of pembrolizumab and glucocorticoids, the symptoms immediately resolved. Regular monitoring of skin, blood tests, and urinalysis are necessary, and the possibility of irAE IgA vasculitis should be considered in cases of purpura and AKI during treatment with immune checkpoint inhibitors.
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Albúminas , Anticuerpos Monoclonales Humanizados , Protocolos de Quimioterapia Combinada Antineoplásica , Carboplatino , Carcinoma de Células Escamosas , Neoplasias Pulmonares , Paclitaxel , Humanos , Masculino , Anciano , Carboplatino/efectos adversos , Carboplatino/administración & dosificación , Carboplatino/uso terapéutico , Paclitaxel/efectos adversos , Paclitaxel/administración & dosificación , Paclitaxel/uso terapéutico , Anticuerpos Monoclonales Humanizados/efectos adversos , Anticuerpos Monoclonales Humanizados/uso terapéutico , Anticuerpos Monoclonales Humanizados/administración & dosificación , Neoplasias Pulmonares/tratamiento farmacológico , Protocolos de Quimioterapia Combinada Antineoplásica/efectos adversos , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Carcinoma de Células Escamosas/tratamiento farmacológico , Albúminas/efectos adversos , Albúminas/administración & dosificación , Vasculitis por IgA/inducido químicamente , Vasculitis por IgA/diagnóstico , Inmunoglobulina A , Inhibidores de Puntos de Control Inmunológico/efectos adversosRESUMEN
BACKGROUND: Air pollution from road traffic is a serious public health problem. Epidemiologic studies have demonstrated adverse health effects associated with environmental pollution. Diesel exhaust is a major contributor to ambient particulate matter air pollution. We studied the effects of exposure to diesel exhaust particles on allergic conjunctivitis using cultured conjunctival epithelial cells obtained from healthy people. OBJECTIVE: To identify the factors involved in the human conjunctival epithelial response to diesel exhaust in vitro. METHODS: Healthy individuals underwent conjunctival biopsy, and the samples were incubated on conjunctival epithelial sheets. We investigated the effects of exposure to diesel exhaust using GeneChip arrays. The adhesion molecules and cytokines showing increased expression on GeneChip arrays were verified by real-time reverse transcription polymerase chain reaction and enzyme-linked immunosorbent assay. RESULTS: The GeneChip array showed increased expression of adhesion molecules, cytokines, chemokines, and growth factors after exposure to diesel exhaust. Real-time reverse transcription polymerase chain reaction and enzyme-linked immunosorbent assay confirmed that the expression of intercellular adhesion molecule 1 and interleukin 6, in particular, were significantly upregulated. CONCLUSION: Our experimental data confirm that exposure to diesel exhaust particles increases inflammatory factor expression in human conjunctiva and thereby contributes to allergic conjunctival responses.
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Conjuntiva/metabolismo , Emisiones de Vehículos/toxicidad , Moléculas de Adhesión Celular/genética , Células Cultivadas , Conjuntiva/citología , Citocinas/genética , Células Epiteliales/metabolismo , Humanos , Análisis de Secuencia por Matrices de Oligonucleótidos , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
BACKGROUND: Eosinophils appear to be key cells in the pathogenesis of conjunctival inflammation in atopic keratoconjunctivitis (AKC). Chemoattractant receptor homologous molecule expressed on TH2 cells (CRTH2) mediates prostaglandin D2 (PGD2)-dependent migration of eosinophils. However, it is unclear whether CRTH2/PGD2-dependent eosinophil migration is upregulated in allergic diseases. OBJECTIVE: To compare the chemotactic responses of peripheral blood eosinophils to prostaglandin D2 in patients with severe AKC and healthy individuals. METHODS: We used an enzyme immunoassay system to measure PGD2 levels in tears and blood samples from healthy individuals and patients with AKC. CRTH2 expression on peripheral blood eosinophils was determined using reverse-transcriptase polymerase chain reaction (RT-PCR), flow cytometry, and an oligonucleotide array system. Chemotaxis experiments were performed using a modified Boyden chamber technique and an optical assay system. RESULTS: The PGD2 concentrations were higher in tears from patients with severe AKC compared with healthy individuals. RT-PCR (severe and mild cases), flow cytometry (mild cases), and GeneChip analyses revealed a significantly higher expression of CRTH2 on peripheral blood eosinophils in patients with AKC than in healthy individuals. PGD2 and its stable metabolite 13,14-dihydro-15-keto-PGD2, a CRTH2 agonist, induced chemotaxis of eosinophils from patients with AKC; chemotaxis was significantly enhanced in eosinophils from patients with severe AKC compared with those from healthy individuals. CONCLUSION: CRTH2 is more abundantly expressed on eosinophils from patients with AKC and promoted PGD2-dependent migration to a greater extent than in healthy individuals.
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Quimiotaxis de Leucocito/fisiología , Eosinófilos/metabolismo , Hipersensibilidad/metabolismo , Queratoconjuntivitis/metabolismo , Prostaglandina D2/metabolismo , Adolescente , Adulto , Niño , Ensayo de Inmunoadsorción Enzimática , Femenino , Citometría de Flujo , Humanos , Masculino , Análisis de Secuencia por Matrices de Oligonucleótidos , Receptores Inmunológicos/biosíntesis , Receptores de Prostaglandina/biosíntesis , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Adulto JovenRESUMEN
BACKGROUND: Chronic rhinosinusitis (CRS) is characterized by local inflammation of the sinonasal tissues. CRS patients with nasal polyps and asthma often develop acute exacerbation of sinonasal symptoms after upper respiratory tract infections. However, the influence of concomitant asthma on the nasal immune response to viral infection remains unclear. METHODS: Specimens of nasal polyp and mucosal tissues were obtained from 3 groups of CRS patients (n = 14 per group): 1) patients without asthma (CRS group), 2) patients with aspirin-tolerant asthma (ATA group), and 3) patients with aspirin-intolerant asthma (AIA group). Nasal fibroblasts isolated from the specimens were stimulated with poly I:C. CXCL10 expression was analyzed by the quantitative real-time polymerase chain reaction and enzyme-linked immunoadsorbent assay. Biopsy specimens from CRS patients without asthma were subjected to immunohistochemistry for detection of T-bet and GATA-3 expression in CD3+ T cells by double labeling. RESULTS: Nasal fibroblasts from the ATA and AIA groups showed significantly enhanced expression of CXCL10 mRNA and protein after poly I:C stimulation compared with cells from the CRS group and the control group (normal nasal mucosa). In addition to T helper (Th)2 cells, there was more abundant infiltration of Th1 cells into tissues from the AIA and ATA groups. CONCLUSIONS: Our findings suggest that CRS associated with asthma may become intractable through the over-production of CXCL10 in response to viral infection.
Asunto(s)
Asma Inducida por Aspirina/inmunología , Asma/inmunología , Quimiocina CXCL10/metabolismo , Pólipos Nasales/inmunología , Senos Paranasales/inmunología , Rinitis/inmunología , Sinusitis/inmunología , Adolescente , Adulto , Anciano , Asma/complicaciones , Asma Inducida por Aspirina/complicaciones , Complejo CD3/metabolismo , Células Cultivadas , Quimiocina CXCL10/genética , Enfermedad Crónica , Resistencia a Medicamentos , Femenino , Fibroblastos/inmunología , Factor de Transcripción GATA3/metabolismo , Humanos , Masculino , Persona de Mediana Edad , Pólipos Nasales/complicaciones , Poli I-C/inmunología , Rinitis/complicaciones , Sinusitis/complicaciones , Proteínas de Dominio T Box/metabolismo , Células TH1/inmunología , Células Th2/inmunología , Regulación hacia Arriba , Adulto JovenRESUMEN
Objectives: To compare vaccinated-side axillary lymph node uptake on 18F-fluorodeoxyglucose (FDG) positron emission tomography/computed tomography (PET/CT) after coronavirus disease-2019 (COVID-19) and influenza vaccination. Methods: We retrospectively analyzed 177 patients who underwent 18F-FDG PET/CT after COVID-19 or influenza vaccination. We compared the uptake of the vaccinated-side axillary lymph nodes of 109 COVID-19 vaccinated patients with those of a lot of influenza-vaccinated patients. We also compared the uptake between 66 patients who received the first COVID-19 vaccination with 43 who received the second COVID-19 vaccination. Results: 18F-FDG-avid axillary lymph nodes on the vaccinated side were significantly more frequently observed in the COVID-19 group (45%) than in the influenza group (19%) (p<0.001). When the interval between vaccination to PET/CT was within 7 days, there was no significant difference in the frequency of 18F-FDG-avid vaccinated-side axillary lymph nodes between the groups (COVID-19 group: 41% vs. influenza group: 45%, p=0.724). When the interval was over 7 days, 18F-FDG-avid lymph nodes were much more frequent in the COVID-19 group (47%) than in the influenza group (7%) (p<0.001). Comparing the first and second COVID-19 groups, 18F-FDG-avid lymph nodes were more frequent in the second vaccination group than in the first vaccination group, but the difference was not significant. Conclusion: 18F-FDG-avid vaccinated-side axillary lymph nodes were more frequently observed in the COVID-19 group than in the influenza group. In the case of the COVID-19 vaccine, a delay of 18F-FDG PET/CT examination is recommended by a longer interval from vaccination than in the influenza vaccine.
RESUMEN
PURPOSE: To date, mouse lacrimal gland epithelial cells have been cultured successfully but only in cases involving newborn mouse lacrimal glands. In this work, we attempted to cultivate and characterize adult mouse lacrimal gland epithelial cells. METHODS: Lacrimal glands were removed from newborn mice (C57B/6) and isolated lacrimal gland epithelial cells were seeded onto tissue culture treated or low adherent culture dishes in Cnt-07 culture medium with or without cholera toxin. Cultivated cells were characterized by immunostaining with pan-cytokeratin, α-smooth muscle actin, and lactoferrin antibodies. Lacrimal gland cells from 7-week-old green fluorescent protein (GFP) and non-GFP (C57B/6) mice were mixed and seeded onto uncoated dishes to assess sphere-forming efficiency. Cells were also seeded onto 3T3 cell feeder layers to assess colony forming efficiency. RESULTS: Lacrimal gland epithelial cells were selectively cultured with cholera toxin, and cell type was verified by pan-cytokeratin and α-smooth muscle actin immunostaining. Sphere formation from single cells of adult mice was observed using specific medium and low adherent culture dishes. These cells could also undergo colony formation on 3T3 feeder cells. CONCLUSIONS: Adult mouse lacrimal gland epithelial cells were successfully cultivated in cholera toxin-containing medium, and were observed to form spheres from single cells.