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Changes in both the social environment (e.g., the increased use of electronic media) and the atmospheric environment (e.g., air pollution and dust) have contributed to an increasing incidence of eye disease and an increased need for eye care. Notably, the signs and symptoms of dry eye syndrome can impact the daily quality of life for various age groups, including the elderly, and usually requires active treatment. The symptoms of dry eye syndrome include tear film instability, hyperosmolarity, ocular surface inflammation and damage, and neurosensory abnormalities. As treatments for dry eye are being developed, a standardized guideline is needed to increase the efficiency of drug development and improve the quality of clinical trial data. In this paper, we present general considerations for the pharmaceutical industry and clinical trial investigators designing clinical trials focused on the development of drugs to treat dry eye syndrome.
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Introduction: As minimal invasive techniques for depressed calcaneal fracture treatment have become more common, great progress has been achieved with various surgical methods. While these techniques are still currently utilized, new methods have been developed. This study aimed to report the radiologic and clinical outcomes of depressed calcaneal fracture treatment with the "push-out molding" technique and to propose its clinical utility. Materials and methods: From March 2009 to October 2020, a retrospective study was conducted with 52 patients, who received the "push-out molding" technique to treat depressed intra-articular calcaneal fractures (Sanders type II, III, IV). Exclusion criteria were as follows: patients with bilateral calcaneal fractures, open fractures, and a follow-up period <12 months. Radiologic parameters were assessed at following periods: preoperative, postoperative, 3-month follow-up, and last follow-up. Limitation of range of motion (ROM), subjective satisfaction, and complications were assessed at the last follow-up period. Repeated measures ANOVA was used to analyze values at preoperative, postoperative, 3-month, and last follow-up periods. Results: Significant differences in the talo-calcaneal angle (p < 0.001), Böhler's angle (p=<0.001), Gissane's angle (p = 0.023), distance from the lower cortical border of calcaneus to the anterior (p=<0.001) and posterior (p=<0.001) points of posterior articular surface, calcaneal length (p = 0.019), and talo-calcaneal height (p=<0.001). Postoperatively, the posterior articular surface was well maintained, while 21.2% retained a ROM limitation by 20° or higher. Subjective satisfaction was as follows: excellent (42.3%), good (48.1%), fair (9.6%), and poor (0%). Conclusion: The "push-out molding" is a simple technique with the advantage of not requiring much force to treat depressed calcaneal fractures. It can be used as a beneficial surgical technique with minimal damage to the soft tissue, owing to the reduction from the depressed interior part and less severe ROM limitation.
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BACKGROUND: Osteomyelitis due to Candida krusei are extremely rare, given that only six cases have been reported, all of which are limited to the patients with immunocompromising risk factors. Here we report a case of C. krusei osteomyelitis in an immunocompetent patient, presenting with multiple huge cystic lesions of talus. CASE PRESENTATION: A 66-year-old female presented with one year history of painful swelling of right ankle and a draining sinus around lateral malleolus. Five months and three months ago, she had undergone arthroscopic synovectomy and bursectomy which revealed no causative organism. Open bursectomy with sinus tract excision was performed and intravenous antibiotic was administered. Two year after the surgery, the patient revisited the clinic for recurrent painful swelling with pus drainage at the same location. Multiple huge cystic lesions with osteolysis and sclerotic rim of talus were found and C. krusei was isolated from tissue culture. The patient received surgical debridement and prolonged antifungal treatment comprising caspofungin and voriconazole. CONCLUSIONS: In this case, C. krusei infection showed atypically aggressive osteolysis shown as multiple huge cystic abscess. High index of suspicion is critical for early diagnosis and treatment to prevent such devastating results even in an immunocompetent patient.
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Osteólisis , Osteomielitis , Astrágalo , Anciano , Antifúngicos/uso terapéutico , Femenino , Humanos , Osteomielitis/complicaciones , Osteomielitis/diagnóstico por imagen , Osteomielitis/cirugía , Pichia , Astrágalo/diagnóstico por imagen , Astrágalo/cirugíaRESUMEN
The global crisis caused by the coronavirus disease 2019 (COVID-19) led to the most significant economic loss and human deaths after World War II. The pathogen causing this disease is a novel virus called the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). As of December 2020, there have been 80.2 million confirmed patients, and the mortality rate is known as 2.16% globally. A strategy to protect a host from SARS-CoV-2 is by suppressing intracellular viral replication or preventing viral entry. We focused on the spike glycoprotein that is responsible for the entry of SARS-CoV-2 into the host cell. Recently, the US Food and Drug Administration/EU Medicines Agency authorized a vaccine and antibody to treat COVID-19 patients by emergency use approval in the absence of long-term clinical trials. Both commercial and academic efforts to develop preventive and therapeutic agents continue all over the world. In this review, we present a perspective on current reports about the spike glycoprotein of SARS-CoV-2 as a therapeutic target.
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BACKGROUND: This study aims to analyze the outcomes of subtalar distraction arthrodesis (SDA) for calcaneal malunion using structural freeze-dried iliac allograft (FDIA) compared to using autologous iliac bone (AIB). METHODS: We retrospectively evaluated 57 consecutive cases (51 patients) of calcaneal malunion between March 2006 and December 2017. All patients were followed for an average of 22.8 months. All cases were treated by SDA using structural FDIA (17 cases, group 1), or AIB (40 cases, group 2). The outcome measures included the American Orthopaedic Foot and Ankle Society ankle-hindfoot (AOFAS) scores, visual analog scale (VAS) pain scores, and radiographic measurements. RESULTS: The mean postoperative 3, 6, and 12 months of AOFAS scores and VAS pain score were significantly better in group 2 than those in group 1 (p < 0.05, for all). There were 3 cases (17.6%) of nonunion in the group 1, whereas the group 2 had 2 cases (5.0%), which did not shown significant difference between two groups (p = 0.492). Although the mean pre-, postoperative, and final follow-up radiologic parameters in both groups were similar, (p > 0.05, for all) the difference of talocalcaneal height, talocalcaneal angle, and talar declination angle from postoperative to final follow-up were significantly bigger in the group 1. (p < 0.05, for all). CONCLUSION: Although union rate was not significantly different between the two groups, we obtained more favorable clinical and radiologic outcomes in the autologous iliac bone group. Using FDIA without any orthobiological agent for SDA, there were significant more loss of radiological parameters due to inferior incorporation and biomechanical properties. When considering the SDA for calcaneal malunion, routine use of FDIA without any orthobiological agents as an interpositional graft for SDA is not recommended.
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Calcáneo , Articulación Talocalcánea , Artrodesis , Calcáneo/diagnóstico por imagen , Calcáneo/cirugía , Humanos , Ilion/diagnóstico por imagen , Ilion/cirugía , Estudios Retrospectivos , Articulación Talocalcánea/diagnóstico por imagen , Articulación Talocalcánea/cirugía , Resultado del TratamientoRESUMEN
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV2) is a positive-sense single-stranded RNA (+ssRNA) that causes coronavirus disease 2019 (COVID-19). The viral genome encodes twelve genes for viral replication and infection. The third open reading frame is the spike (S) gene that encodes for the spike glycoprotein interacting with specific cell surface receptor - angiotensin converting enzyme 2 (ACE2) - on the host cell membrane. Most recent studies identified a single point mutation in S gene. A single point mutation in S gene leading to an amino acid substitution at codon 614 from an aspartic acid 614 into glycine (D614G) resulted in greater infectivity compared to the wild type SARS-CoV2. We were interested in investigating the mutation region of S gene of SARS-CoV2 from Korean COVID-19 patients. New mutation sites were found in the critical receptor binding domain (RBD) of S gene, which is adjacent to the aforementioned D614G mutation residue. This specific sequence data demonstrated the active progression of SARS-CoV2 by mutations in the RBD of S gene. The sequence information of new mutations is critical to the development of recombinant SARS-CoV2 spike antigens, which may be required to improve and advance the strategy against a wide range of possible SARS-CoV2 mutations.
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BACKGROUND: Bunionette deformity is a painful bony prominence of the 5th metatarsal. We evaluated outcomes of using a Kramer osteotomy to treat this condition. METHODS: Retrospective study of patients treated with a Kramer osteotomy from 2003 and 2016. Outcome measures included Foot Functional Index (FFI) and radiographic measurements. RESULTS: 38 patients (43 feet) with an average follow-up of 55 months. Mean postoperative FFI1 was 19.4. Mean 4-5 IMA2 improved 3.9°, from 8.3° preoperatively to 4.4° on final postoperative films (p<0.01). Mean MTP-53 angle improved 13.2° from 13.6° preoperatively to 0.4° at final follow-up (p<0.01). There were 5 delayed unions (11.6%) and 1 non-union (2.3%). CONCLUSIONS: The Kramer osteotomy is an effective treatment option in patients with bunionette deformity, with significant correction of the 4-5 IM2 and MTP-53 angles and few complications.
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Juanete de Sastre/diagnóstico por imagen , Juanete de Sastre/cirugía , Osteotomía/métodos , Femenino , Humanos , Masculino , Huesos Metatarsianos/cirugía , Persona de Mediana Edad , Estudios Retrospectivos , Resultado del TratamientoRESUMEN
BACKGROUND: There is no consensus on the optimal treatment or preferred method of operation for the management of acute deltoid ligament injuries during an ankle fracture fixation. This study aimed to analyze the outcomes of repairing the deltoid ligament during the fixation of an ankle fracture compared to conservative management. METHODS: We retrospectively evaluated 78 consecutive cases of a ruptured deltoid ligament with an associated ankle fracture between 2001 and 2016. All of the ankle fractures were treated with a plate and screw fixation. Patients in the conservative treatment for ruptured deltoid ligament underwent management from 2001 to 2008 (37 fractures, group 1), while the operative treatment for ruptured deltoid ligament was included from 2009 to 2016 (41 fractures, group 2). The outcome measures included radiographic findings, the American Orthopaedic Foot & Ankle Society ankle-hindfoot scores, visual analog scale scores, and the Foot Function Index. All patients were followed for an average of 17 months. RESULTS: Radiologic findings in both groups were comparable, but the final follow-up of the medial clear space (MCS) was significantly smaller in the group 2 ( P < .01). Clinical outcomes were similar between the two groups ( P > .05). Comparing those who underwent syndesmotic fixation between both groups, group 2 showed a significantly smaller final follow-up MCS, and all clinical outcomes were better in group 2 ( P < .05). Linear regression analysis showed that the final follow-up MCS had a significant influence on clinical outcomes ( P < .05). CONCLUSION: Although the clinical outcomes were not significantly different between the 2 groups, we obtained a more favorable final follow-up MCS in the deltoid repair group. Particularly when accompanied by a syndesmotic injury, the final follow-up MCS and the clinical outcomes were better in the deltoid repair group. In the case of high-grade unstable fractures of the ankle with syndesmotic instability, a direct repair of the deltoid ligament was adequate for restoring medial stability. LEVEL OF EVIDENCE: Level III, retrospective comparative case series.
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Fracturas de Tobillo/cirugía , Articulación del Tobillo/cirugía , Fijación Interna de Fracturas/métodos , Rotura/fisiopatología , Traumatismos del Tobillo/cirugía , Tornillos Óseos , Humanos , Ortopedia , Estudios RetrospectivosRESUMEN
BACKGROUND: An infected Achilles tendon after tendon repair is particularly difficult to treat because of the poor vascularity of the tendon as well as the thin surrounding soft tissue. For treatment of an infected Achilles tendon following tendon repair, we first focused on complete debridement and then promoted fibrous scar healing of the Achilles tendon using functional treatment. METHODS: We retrospectively reviewed all of the medical records of 15 tertiary referral patients with postoperative infection of the Achilles tendon occurring between 2007 and 2012. The mean follow-up time was 33 months (range, 22 to 97 months). The infected tissue and the necrotic tendon were debrided, and the ankle was placed in a short leg splint for 2 weeks. The splint was then replaced with an ankle brace for the next 4 weeks. Partial weight-bearing was allowed immediately, and full weight-bearing was allowed at 2 weeks postoperatively. We assessed and recorded the physical parameters such as the range of motion, calf circumference, ability to perform a single-limb heel rise, patient satisfaction, and Arner-Lindholm scale. Laboratory tests, postoperative ultrasonography, and isokinetic plantar flexion power tests were also performed. RESULTS: At a mean time of 17 days (range, 8 to 30 days) after debridement, infection signs such as discharge from the wound, redness, and local warmth resolved. The wound had healed and the stitches were removed at a mean of 17 days following the wound repair. At the time of the latest follow-up, there were no signs of active infection. Achilles tendon continuity recovered in all patients by fibrous scar healing. Compared with the contralateral side, there was no difference in the ankle range of motion in 8 patients. According to the Arner-Lindholm scale, 9 of the 15 results were excellent and 6 were good. Ten patients were able to perform a single-limb heel rise. Eleven of 15 patients returned to their pre-injury recreational activities. Diffuse homogeneous echotexture of the Achilles tendon with continuity was observed on the ultrasonographic examination. CONCLUSIONS: In this retrospective series, radical debridement, combined with antibiotic therapy and functional rehabilitation, was successful in eradicating infection and maintaining function in patients with postoperative infection following Achilles tendon repair. LEVEL OF EVIDENCE: Therapeutic Level IV. See Instructions for Authors for a complete description of levels of evidence.
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Tendón Calcáneo/cirugía , Procedimientos Ortopédicos/efectos adversos , Rotura/cirugía , Infecciones Estafilocócicas/cirugía , Traumatismos de los Tendones/cirugía , Tendón Calcáneo/microbiología , Adulto , Anciano , Desbridamiento/métodos , Femenino , Humanos , Masculino , Persona de Mediana Edad , Procedimientos Ortopédicos/métodos , Complicaciones Posoperatorias/microbiología , Complicaciones Posoperatorias/rehabilitación , Complicaciones Posoperatorias/cirugía , Estudios Retrospectivos , Rotura/rehabilitación , Infecciones Estafilocócicas/etiología , Infecciones Estafilocócicas/rehabilitación , Traumatismos de los Tendones/rehabilitación , Resultado del Tratamiento , Soporte de Peso/fisiologíaRESUMEN
Although it has been established that diabetes increases susceptibility to infections, the role of insulin (INS) in the immune response is unknown. Here, we investigated the immunological function of INS. Proinsulin dimer (pINSd) was a potent immune stimulus that induced inflammatory cytokines, but mature INS was unable to induce an immune response. An affinity-purified rabbit polyclonal antibody raised against mature IL-1α recognized IL-1α and pINS but failed to detect mature INS and IL-1ß. Analysis of the pINS sequence revealed the existence of an INS/IL-1α motif in the C-peptide of pINS. Surprisingly, the INS/IL-1α motif was recognized by monoclonal antibody raised against IL-1α. Deleting the INS/IL-1α motif in pINSd and IL-1α changed their activities. To investigate the pINSd receptor, the reconstitution of IL-1 receptor 1 (IL-1R1) in Wish cells restored pINSd activity that was reversed by an IL-1R antagonist. These data suggested that pINSd needs IL-1R1 for inflammatory cytokine induction. Mouse embryo fibroblast cells of IL-1R1-deficient mice further confirmed that pINSd promotes immune responses through IL-1R1.
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Citocinas/biosíntesis , Mediadores de Inflamación/metabolismo , Interleucina-1alfa/metabolismo , Proinsulina/metabolismo , Receptores de Interleucina-1/metabolismo , Animales , Células Cultivadas , Interleucina-1alfa/química , Ratones , Proinsulina/químicaRESUMEN
Interleukin (IL)-33 is an inflammatory cytokine and belongs to the IL-1 family of cytokines. There are eleven members of the IL-1 family of cytokines and all have important roles in host defense against infections. Their levels are increased during infection and in various auto-inflammatory diseases. IL-33 is also associated with autoimmune diseases such as asthma, atopic dermatitis, rheumatoid arthritis, and atherosclerosis. IL-33 receptors consist of IL-1R4 and IL-1R3 to induce both Th1 and Th2 type immune response. Here we present the development of monoclonal antibodies (mAbs) against human mature IL-33. Recombinant human mature IL-33 protein was expressed in E. coli and purified by multi-step affinity chromatography. The human IL-33 activity was examined in HMC-1 and Raw 264.7 cells. Mice were immunized with the biologically active mature IL-33 to generate mAb against IL-33. The anti-IL-33 mAb (clone/4) was used as a capture antibody for a sandwich enzyme-linked immunosorbent assay (ELISA). This assay detects mature IL-33 with a high sensitivity (80 pg/mL) but does not recognize the biologically inactive precursor IL-33. This article describes the methods for a newly developed IL-33 ELISA kit that is specific for mature IL-33 and may be used to analyze bioactive mature IL-33 in various immunological diseases.
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Ensayo de Inmunoadsorción Enzimática/métodos , Interleucina-33/química , Interleucina-33/inmunología , Animales , Anticuerpos Monoclonales/inmunología , Reacciones Antígeno-Anticuerpo , Línea Celular , Humanos , Interleucina-33/genética , Ratones , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
Interleukin-33 (IL-33) receptors are composed of ST2 (also known as IL-1R4), a ligand binding chain, and IL-1 receptor accessory protein (IL-1RAcP, also known as IL-1R3), a signal transducing chain. IL-1R3 is a common receptor for IL-1α, and IL-1ß, IL-33, and three IL-36 isoforms. A549 human lung epithelial cells are highly sensitive to IL-1α and IL-1ß but not respond to IL-33. The lack of responsiveness to IL-33 is due to ST2 expression. ST2 was stably transfected into A549 cells to reconstitute its activity. RT-PCR and FACS analysis confirmed ST2 expression on the cell surface of A549/ST2 cells. Upon IL-33 stimulation, A549/ST2 cells induced IL-8 and IL-6 production in a dose dependent manner while A549/mock cells remained unresponsive. There was no difference in IL-1α and IL-1ß activity in A549/ST2 cells compared to A549/mock cells despite the fact that IL-33 shares IL-1R3 with IL-1α/ß. IL-33 activated inflammatory signaling molecules in a time- and dose-dependent manner. Anti-ST2 antibody and soluble recombinant ST2-Fc abolished IL-33-induced IL-6 and IL-8 production in A549/ST2 cells but the IL-1 receptor antagonist failed to block IL-33-induced cytokines. This result demonstrates for the first time the reconstitution of ST2 in A549 human lung epithelial cell line and verified its function in IL-33-mediated cytokine production and signal transduction.
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Proteína Antagonista del Receptor de Interleucina 1/metabolismo , Proteína 1 Similar al Receptor de Interleucina-1/metabolismo , Interleucina-1alfa/metabolismo , Interleucina-1beta/metabolismo , Interleucina-33/metabolismo , Transducción de Señal/fisiología , Células A549 , Regulación de la Expresión Génica/fisiología , Células Endoteliales de la Vena Umbilical Humana , Humanos , Proteína Antagonista del Receptor de Interleucina 1/genética , Proteína 1 Similar al Receptor de Interleucina-1/genética , Interleucina-1alfa/genética , Interleucina-1beta/genética , Interleucina-33/genética , Interleucina-6/sangre , Interleucina-6/genética , Interleucina-8/biosíntesis , Interleucina-8/genéticaRESUMEN
PURPOSE: This study was designed to evaluate the mid-term results and efficacy of subtalar distraction double bone-block arthrodesis for calcaneal malunion. MATERIALS AND METHODS: From January 2004 to June 2007, we operated on 6 patients (10 cases). There were 5 males (9 cases) and 1 female (1 case), four of which presented with bilateral calcaneal malunion. Seven cases were operated on initially. The period between initial injury and arthrodesis was 23 months, and the average follow up period was 58 months. In operation, we applied an extensile lateral approach and arthrodesis was performed through a tricortical double bone-block and cannulated screws. The American Orthopaedic Foot and Ankle Society (AOFAS) Ankle-Hindfoot scale was used for clinical evaluation. In radiologic analysis, plain X-ray and CT were examined to assess union and various parameters. RESULTS: The mean age of the patients was 41 years. All cases achieved radiologic union at the final follow-up. The mean AOFAS Ankle-Hindfoot scale (maximum of 94 points) increased from 43.3 points preoperatively to 85.4 points at the final follow-up. The radiologic analysis of the pre- and postoperative standing lateral radiographs showed improvements of 5.6 mm in talo-calcaneal height, 1.8° in talocalcaneal angle, 5.1° in talar declination angle and 5.3° in talo-first metatarsal angle. CONCLUSION: Subtalar distraction two bone-block arthrodesis provides overall good results not only in the short term but also the mid-term with significant improvement in clinical and radiologic outcomes. This procedure warrants consideration for managing calcaneal malunion with loss of height and subtalar arthritis.
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Calcáneo/lesiones , Fracturas Mal Unidas/cirugía , Adulto , Artrodesis/métodos , Calcáneo/cirugía , Femenino , Humanos , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , Resultado del TratamientoRESUMEN
Interleukin-32 (IL-32) is a cytokine and inducer of various proinflammatory cytokines such as TNFα, IL-1ß, and IL-6 as well as chemokines. There are five splicing variants (α, ß, γ, delta, and epsilon) and IL-32γ is the most active isoform. We generated human IL-32γ transgenic (IL-32γ TG) mice to express high level of IL-32γ in various tissues, including immune cells. The pathology of sepsis is based on the systemic inflammatory response that is characterized by upregulating inflammatory cytokines in whole body, particularly in response to gram-negative bacteria. We investigated the role of IL-32γ in a mouse model of experimental sepsis by using lipopolysaccharides (LPS). We found that IL-32γTG mice resisted LPS-induced lethal endotoxemia. IL-32γ reduced systemic cytokines release after LPS administration but not the local immune response. IL-32γTG increased neutrophil influx into the initial foci of the primary injected site, and prolonged local cytokines and chemokines production. These results suggest that neutrophil recruitment in IL-32γTG occurred as a result of the local induction of chemokines but not the systemic inflammatory cytokine circulation. Together, our results suggest that IL-32γ enhances an innate immune response against local infection but inhibits the spread of immune responses, leading to systemic immune disorder.
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Expresión Génica , Interleucinas/metabolismo , Lipopolisacáridos/toxicidad , Choque Séptico/inducido químicamente , Choque Séptico/prevención & control , Síndrome de Respuesta Inflamatoria Sistémica/inducido químicamente , Síndrome de Respuesta Inflamatoria Sistémica/prevención & control , Animales , Modelos Animales de Enfermedad , Resistencia a la Enfermedad , Humanos , Interleucinas/genética , Lipopolisacáridos/administración & dosificación , Ratones , Ratones Transgénicos , Choque Séptico/inmunología , Síndrome de Respuesta Inflamatoria Sistémica/inmunologíaRESUMEN
α1-Antitrypsin (AAT) is a member of the serine proteinase inhibitor family that impedes the enzymatic activity of serine proteinases, including human neutrophil elastase, cathepsin G and neutrophil proteinase 3. Here, we expressed recombinant AAT by fusing the intact AAT gene to the constant region of IgG1 to generate soluble recombinant AAT-Fc protein. The recombinant AAT-Fc protein was produced in Chinese hamster ovary (CHO) cells and purified using mini-protein A affinity chromatography. Recombinant AAT-Fc protein was tested for antiinflammatory function and AAT-Fc sufficiently suppressed tumor necrosis factor (TNF)-α-induced interleukin (IL)-6 in human peripheral blood mononuclear cells (PBMCs) and inhibited cytokine-induced TNFα by different cytokines in mouse macrophage Raw 264.7 cells. However, AAT-Fc failed to suppress lipopolysaccharide-induced cytokine production in both PBMCs and macrophages. In addition, our data showed that AAT-Fc blocks the development of hyperglycemia in a streptozotocin-induced mouse model of diabetes. Interestingly, we also found that plasma-derived AAT specifically inhibited the enzymatic activity of elastase but that AAT-Fc had no inhibitory effect on elastase activity.
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Antiinflamatorios/farmacología , Inmunoglobulina G/farmacología , Proteínas Recombinantes de Fusión/farmacología , alfa 1-Antitripsina/farmacología , Animales , Antiinflamatorios/uso terapéutico , Glucemia/análisis , Células CHO , Línea Celular , Células Cultivadas , Cricetinae , Cricetulus , Citocinas/farmacología , Diabetes Mellitus Experimental/tratamiento farmacológico , Diabetes Mellitus Experimental/metabolismo , Femenino , Humanos , Fragmentos Fc de Inmunoglobulinas/genética , Fragmentos Fc de Inmunoglobulinas/farmacología , Fragmentos Fc de Inmunoglobulinas/uso terapéutico , Inmunoglobulina G/genética , Inmunoglobulina G/uso terapéutico , Interleucina-6/metabolismo , Leucocitos Mononucleares/efectos de los fármacos , Leucocitos Mononucleares/metabolismo , Ratones , Ratones Endogámicos BALB C , Elastasa Pancreática/metabolismo , Proteínas Recombinantes de Fusión/uso terapéutico , Factor de Necrosis Tumoral alfa/metabolismo , alfa 1-Antitripsina/genética , alfa 1-Antitripsina/uso terapéuticoRESUMEN
IL-33 (IL-1F11) is a member of IL-1 family ligand, which stimulates the production of inflammatory cytokines. IL-33 receptor complex is comprised of IL-1 receptor accessory protein (IL-1RAcP) and ST2 that are activated by IL-33 ligand binding. ST2 is a ligand-binding chain of the IL-33 receptor component, and the soluble ST2 form possesses antagonistic activity. Here, we expressed the extracellular domain of ST2-fused to the immunoglobulin of IgG1 constant region in order to generate a soluble recombinant Fc-ST2. Human and mouse recombinant Fc-ST2 protein were expressed in Chinese hamster ovary cells and purified using a mini-protein A affinity chromatography. The recombinant Fc-ST2 protein was used to examine inhibitory function in IL-33-induced cytokine production in different cell types. The human Fc-ST2 abolished IL-33-induced IL-8 production in human mast cells, but mouse Fc-ST2 failed to inhibit IL-33-induced TNFα production in mouse Raw 264.7 macrophage cells. We further investigated the expression of IL-33 receptor component with various cell lines. IL-33 receptors expression pattern and Fc-ST2 inhibitory activity in different cell types suggest that IL-1RAcP and ST2 are necessary but insufficient for IL-33 activity. Our results suggest that an additional receptor component may participate in the biological activity of IL-33.
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Fragmentos Fc de Inmunoglobulinas/metabolismo , Proteína Accesoria del Receptor de Interleucina-1/inmunología , Interleucinas/inmunología , Macrófagos/efectos de los fármacos , Mastocitos/efectos de los fármacos , Receptores de Superficie Celular/inmunología , Receptores de Interleucina/inmunología , Animales , Células COS , Chlorocebus aethiops , Cromatografía de Afinidad , Cricetinae , Humanos , Fragmentos Fc de Inmunoglobulinas/genética , Proteína 1 Similar al Receptor de Interleucina-1 , Interleucina-33 , Interleucina-8/metabolismo , Macrófagos/inmunología , Mastocitos/inmunología , Ratones , Receptores de Superficie Celular/genética , Receptores de Interleucina/genética , Proteínas Recombinantes de Fusión/genética , Transducción de Señal/efectos de los fármacos , Transducción de Señal/inmunología , Especificidad de la Especie , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
The pro-inflammatory cytokines tumor necrosis factor-α (TNFα) and interleukin (IL)-1ß are crucial mediators involved in chronic inflammatory diseases. Inflammatory signal pathways regulate inflammatory cytokine expression-mediated by p38 mitogen activated protein kinase (p38MAPK). Therefore, considerable attention has been given to p38MAPK as a target molecule for the development of a novel anti-inflammatory therapeutics. BIRB 796, one of p38MAPK inhibitor, is a candidate of therapeutic drug for chronic inflammatory diseases. In this study, we investigated the effect of BIRB 796 on inflammatory cytokine productions by lipopolysaccharide (LPS) in different immune cell types. BIRB 796 reduced LPS-mediated IL-8 production in THP-1 cells but not in Raw 264.7 cells. Further analysis of signal molecules by western blot revealed that BIRB 796 sufficiently suppressed LPS-mediated phosphorylation of p38MAPK in both cell types whereas it failed to block inhibitor of kappa B (I-κB) degradation in Raw 264.7 cells. Taken together, these results suggest that the anti-inflammatory function of BIRB 796 depends on cell types.
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OBJECTIVE: To evaluate the role of IL-32 in granulomatosis with polyangiitis (GPA) patients and the relationship between IL-32 and disease activity, as PR3 has the ability to bind and activate IL-32, which has been described as a novel cytokine that induces inflammatory cytokines. METHODS: We investigated the level of IL-32, PR3, TNF-α and IL-6 in GPA patients by using ELISA. Northern blot was used to analyse the level of IL-32 mRNA in leucocytes of GPA patients. The intracellular colocalization of IL-32 and PR3 in leucocytes was examined by IF staining. RESULTS: We observed that IL-32 and PR3 levels in GPA patients were increased significantly when compared with normal individuals and each was tightly associated (P < 0.001). Northern blot analysis revealed that the mRNA level of IL-32 was prominently elevated in leucocytes of GPA patients. The intracellular colocalization of IL-32 and PR3 in leucocytes from GPA patients vs normal individuals was verified by IF staining. CONCLUSION: IL-32 level was elevated in GPA patients but its level was changed by treatment response. IL-32 could be an index in GPA and play a role in the aetiology of GPA.
Asunto(s)
Granulomatosis con Poliangitis/etiología , Interleucinas/fisiología , Adulto , Anciano , Northern Blotting , Ensayo de Inmunoadsorción Enzimática , Femenino , Granulomatosis con Poliangitis/metabolismo , Humanos , Interleucina-6/metabolismo , Interleucinas/metabolismo , Leucocitos/metabolismo , Masculino , Persona de Mediana Edad , Mieloblastina/metabolismo , ARN Mensajero/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo , Vasculitis/etiología , Vasculitis/metabolismoRESUMEN
Interferons (IFNs) are commonly grouped into type I and type II IFN. Type I IFNs are known as antiviral IFNs including IFN-α, IFN-ß, and IFN-ω whereas type II IFN is referred to immune IFN and IFN-γ is only member of the type II IFN. Type I IFNs are induced by virus invading however type II IFN is produced by mitogenic or antigenic stimuli. IFN-τ was first identified in ruminant ungulates as a pregnancy recognition hormone, trophoblastin. IFN-τ constitutes a new class of type I IFN, which possesses the common features of type I IFN, such as the ability to prevent viral infection and to limit cell proliferation. In addition, IFN-τ is unique in that it is induced by pregnancy unlike other type I IFNs. We cloned Bos taurus (B. T.) Coreanae IFN-τ from peripheral blood mononuclear cells. The amino acid sequence of B. T. Coreanae IFN-τ shares only 90.3% identity with that of Holstein dairy cow. Recombinant B. T. Coreanae and Holstein IFN-τ proteins were expressed in Escherichia coli and the antiviral activity of IFN-τ proteins were examined. Both recombinant proteins were active and protected human WISH and bovine MDBK cells from the cytopathic effect of vesicular stomatitis virus. The recombinant IFN-τ protein of B. T. Coreanae and Holstein properly induced the expression of antiviral genes including 2',5'-oligoadenylate synthetase (OAS) and Mx GTPase 1 (Mx-1).
Asunto(s)
Aminoácidos/química , Bovinos/metabolismo , Interferón Tipo I/química , Proteínas Gestacionales/química , 2',5'-Oligoadenilato Sintetasa/metabolismo , Secuencia de Aminoácidos , Animales , Antivirales/farmacología , Secuencia de Bases , Células Sanguíneas/efectos de los fármacos , Células Sanguíneas/inmunología , Línea Celular , Clonación Molecular , Efecto Citopatogénico Viral/efectos de los fármacos , Perros , GTP Fosfohidrolasas/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Interferón Tipo I/genética , Interferón Tipo I/metabolismo , Interferón Tipo I/farmacología , Datos de Secuencia Molecular , Proteínas Gestacionales/genética , Proteínas Gestacionales/metabolismo , Proteínas Gestacionales/farmacología , ARN Mensajero/genética , ARN Mensajero/metabolismo , Proteínas Recombinantes/aislamiento & purificación , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/farmacologíaRESUMEN
Histone modifications are important components of transcriptional regulation and chromatin-based regulatory processes. In addition, WD40-repeat protein and several other components are involved in these functions. Here we present the development of monoclonal antibodies (MAbs) against Arabidopsis HOS15, a WD40-repeat protein. Mice were immunized with recombinant HOS15 (rHOS15) protein for generating MAbs via the classic hybridoma production technique. We confirmed the specific activity of anti-HOS15 MAbs by tobacco transient expression assays. Based on immunoprecipitation assays, the anti-HOS15 MAb was able to detect endogenous HOS15 in Arabidopsis wild-type plants, but not in hos15 mutant plants. Finally, the anti-HOS15 MAbs are highly sensitive for detecting endogenous HOS15 protein. They can be used for immunological and immunoprecipitation assays to support other experimental strategies. In particular, the anti-HOS15 MAbs will be essential tools to investigate the role of HOS15 in the regulation of tolerance to environmental stresses in plants.