PURPOSE: In the Phase III SIROCCO trial (NCT01928771), benralizumab significantly reduced asthma exacerbations and improved lung function and symptoms for patients with severe, uncontrolled eosinophilic asthma. The aim of this subgroup analysis was to evaluate efficacy and safety of benralizumab for Korean patients in SIROCCO. METHODS: SIROCCO was a randomized, double-blind, parallel-group, placebo-controlled trial of 1,204 patients aged 12–75 years with severe asthma uncontrolled by high-dosage inhaled corticosteroids/long-acting β2-agonists (ICS/LABA). Patients received benralizumab 30 mg every 4 weeks (Q4W) or every 8 weeks (Q8W; first 3 doses Q4W) or placebo Q4W for 48 weeks. The primary analysis population comprised patients with blood eosinophil counts ≥ 300 cells/µL. This subgroup analysis evaluated Korean patients from this group. RESULTS: Of 122 Korean patients randomized, 86 had blood eosinophil counts ≥ 300 cells/µL. Benralizumab reduced the annual asthma exacerbation rate by 70% (Q4W: rate estimate 0.79, rate ratio 0.30 [95% confidence interval {CI}, 0.13–0.65], nominal P = 0.003; n = 28) and 85% (Q8W: rate estimate 0.40, rate ratio 0.15 [95% CI, 0.06–0.36], nominal P < 0.001; n = 30) vs. placebo (rate estimate 2.67, n = 28). Prebronchodilator forced expiratory volume in 1 second was increased with benralizumab treatment by 0.270 L (Q4W: 95% CI, 0.039–0.500, nominal P = 0.023; n = 28) and 0.362 L (Q8W: 95% CI, 0.143–0.582, nominal P = 0.002; n = 30) vs. placebo (n = 27). Total asthma symptom score was similar for patients receiving either benralizumab Q4W (−0.27 [95% CI, −0.83 to 0.30], nominal P = 0.356; n = 27) or benralizumab Q8W (0.10 [95% CI, −0.44 to 0.65], nominal P = 0.708; n = 30) vs. placebo (n = 28). Drug-related adverse events were experienced by 2%, 8%, and 5% of patients in the placebo, benralizumab Q4W, and benralizumab Q8W arms. CONCLUSIONS: Benralizumab reduced annual asthma exacerbation rates, increased lung function, and was well-tolerated by Korean patients with severe, uncontrolled eosinophilic asthma.
Arm ; Asthma ; Eosinophils ; Forced Expiratory Volume ; Humans ; Korea ; Lung ; Receptors, Interleukin-5

BACKGROUND: Androgenic alopecia (AGA) is the most common type of hair loss. It is likely inherited genetically and is promoted by dihydrotestosterone. 5α-reductase has been proven a good target through finasteride use. However, the pathogenesis of AGA cannot be fully explained based only on dihydrotestosterone levels. OBJECTIVE: To identify similar hairloss inhibition activity of RE-ORGA with mode of action other than finasteride. METHODS: We prepared RE-ORGA from Korean herb mixtures. We performed MTT assays for cytotoxicity, Cell Counting Kit-8 assays for cell proliferation, and western blot to identify expression levels of 5α-reductase and Bax. RNA-sequencing was performed for the expression patterns of genes in dihydrotestosterone-activated pathways. Anti-inflammatory activity was also assessed by the expression levels of tumor necrosis factor-alpha (TNF-α) and interleukin 6. RESULTS: REORGA could promote the proliferation of human dermal papilla cells and showed low cytotoxicity. It also inhibited the expression of 5α-reductases and Bax in the cells. RNA-sequencing results verified that the mRNA expressions of SRD5A1, Bax, transforming growth factor-beta 1 (TGF-β1), and TGF-β1 induced transcript 1 (TGFβ1I1) were decreased, whereas expression of protein tyrosine kinase 2 beta (PTK2β) was more elevated. REORGA also showed anti-inflammatory activity through decreased mRNA levels of TNF-α. CONCLUSION: Transcriptionally, up-regulation of PTK2β and concomitant down-regulation of TGFβ1I1 imply that RE-ORGA can modulate androgen receptor sensitivity, decreasing the expression of 5α-reductase type II and Bax together with TGF-β1 transcripts; RE-ORGA also showed partial anti-inflammatory activity. Overall, RE-ORGA is expected to alleviate hair loss by regulating 5α-reductase activity and the receptor's androgen sensitivity.
Alopecia ; Blotting, Western ; Cell Count ; Cell Proliferation ; Cholestenone 5 alpha-Reductase ; Dihydrotestosterone ; Down-Regulation ; Finasteride ; Hair ; Humans ; Interleukin-6 ; Protein-Tyrosine Kinases ; Receptors, Androgen ; RNA, Messenger ; Tumor Necrosis Factor-alpha ; Up-Regulation

BACKGROUND: Efficacy and safety of tiotropium bromide, a muscarinic receptor antagonist, in treatment of asthma have been reported. However, its effect on airway remodeling in chronic asthma of the elderly has not been clearly verified. The objective of this study was to investigate the effect of tiotropium and expression of muscarinic receptors as its related mechanism in an aged mouse model of chronic asthma with airway remodeling. METHODS: BALB/c female mice age 6 weeks, 9 and 15 months were sensitized and challenged with ovalbumin (OVA) for three months. Tiotropium bromide was administered during the challenge period. Airway hyperresponsiveness (AHR) and pulmonary inflammation were measured. Parameters of airway remodeling, and expression levels of M2 and M3 receptors were examined. RESULTS: Total cell with eosinophils, increased in the OVA groups by age, was decreased significantly after treatment with tiotropium bromide, particularly in the age group of 15 months. AHR and levels of interleukin (IL)-4, IL-5, and IL-13 were decreased, after tiotropium administration. In old aged group of 9- and 15-months-treated groups, hydroxyproline contents and levels of α-smooth muscle actin were attenuated. Tiotropium enhanced the expression of M2 but decreased expression of M3 in all aged groups of OVA. CONCLUSION: Tiotropium bromide had anti-inflammatory and anti-remodeling effects in an aged mouse model of chronic asthma. Its effects seemed to be partly mediated by modulating expression M3 and M2 muscarinic receptors. Tiotropium may be a beneficial treatment option for the elderly with airway remodeling of chronic asthma.
Actins ; Aged ; Airway Remodeling ; Animals ; Asthma* ; Eosinophils ; Female ; Humans ; Hydroxyproline ; Interleukin-13 ; Interleukin-5 ; Interleukins ; Mice* ; Ovalbumin ; Ovum ; Pneumonia ; Receptors, Muscarinic* ; Tiotropium Bromide*

Asthma, remains symptomatic despite ongoing treatment with high doses of inhaled corticosteroids (ICS) in conjunction with long-acting beta-agonists (LABA), is classified as “severe” asthma. In the course of caring for those patients diagnosed with severe asthma, stepping up from ICS/LABA to more aggressive therapeutic measures would be justified, though several aspects have to be checked in advance (including inhaler technique, adherence to therapy, and possible associated comorbidities). That accomplished, it would be advisable to step up care in accordance with the Global Initiative for Asthma (GINA) recommendations. Possible strategies include the addition of a leukotriene receptor antagonist or tiotropium (to the treatment regimen). The latter has been shown to be effective in the management of several subgroups of asthma. Oral corticosteroids have commonly been used for the treatment of patients with severe asthma in the past; however, the use of oral corticosteroids is commonly associated with corticosteroid-related adverse events and comorbidities. Therefore, according to GINA 2017 these patients should be referred to experts who specialize in the treatment of severe asthma to check further therapeutic options including biologics before starting treatment with oral corticosteroids.
Adrenal Cortex Hormones ; Asthma ; Biological Products ; Comorbidity ; Humans ; Immunoglobulin E ; Interleukin-5 ; Nebulizers and Vaporizers ; Receptors, Leukotriene ; Tiotropium Bromide

BACKGROUND/AIMS: Inhaled corticosteroids are the most effective treatment currently available for asthma, but their beneficial effect against airway remodeling is limited. The tyrosine kinase inhibitor nilotinib has inhibitory activity against c-kit and the platelet-derived growth factor receptor. We compared the effects of fluticasone and nilotinib on airway remodeling in a chronic asthma model. We also examined whether co-treatment with nilotinib and fluticasone had any synergistic effect in preventing airway remodeling. METHODS: We developed a mouse model of airway remodeling, including smooth muscle thickening, in which ovalbumin (OVA)-sensitized female BALB/c-mice were repeatedly exposed to intranasal OVA administration twice per week for 3 months. Mice were treated with fluticasone and/or nilotinib intranasally during the OVA challenge. RESULTS: Mice chronically exposed to OVA developed eosinophilic airway inflammation and showed features of airway remodeling, including thickening of the peribronchial smooth muscle layer. Both fluticasone and nilotinib attenuated airway smooth muscle thickening. However, only nilotinib suppressed fibrotic changes, demonstrating inhibition of collagen deposition. Fluticasone reduced pro-inflammatory cells, such as eosinophils, and several cytokines, such as interleukin 4 (IL-4), IL-5, and IL-13, induced by repeated OVA challenges. On the other hand, nilotinib reduced transforming growth factor β1 levels in bronchoalveolar lavage fluid and inhibited fibroblast proliferation significantly. CONCLUSIONS: These results suggest that fluticasone and nilotinib suppressed airway remodeling in this chronic asthma model through anti-inflammatory and anti-fibrotic pathways, respectively.
Adrenal Cortex Hormones ; Airway Remodeling ; Animals ; Asthma* ; Bronchoalveolar Lavage Fluid ; Collagen ; Cytokines ; Eosinophils ; Female ; Fibroblasts ; Fluticasone* ; Hand ; Humans ; Inflammation ; Interleukin-13 ; Interleukin-4 ; Interleukin-5 ; Mice ; Muscle, Smooth ; Ovalbumin ; Ovum ; Protein-Tyrosine Kinases ; Receptors, Platelet-Derived Growth Factor ; Transforming Growth Factors

OBJECTIVETo study the effect of Wenyang Decoction (WD) on the differentiation of CD34+ progenitor cells of occupational asthma (OA) model rats.

METHODSFifty healthy male SD rats were randomly divided into five groups, i.e., the model group, the blank control group,the WD group,the Western medicine group,the combined group, 10 in each group. Prednisone suspension (10 mg/kg) was administered to rats in the Western medicine group by gastrogavage. WD (20 g/kg) was administered to rats in the WD group by gastrogavage. Prednisone suspension plus WD was administered to rats in the combined group by gastrogavage. Normal saline was administered to rats in the model group and the blank control group by gastrogavage. The general condition of rats was observed. Expression levels of peripheral blood IL-5 and eotaxin, eosinophils (EOS), CD34+, CC chemokine receptor 3 (CCR3+) in bone marrow suspension were detected by ELISA, Wirght-Giemsa, and flow cytometry, respectively.

RESULTSCompared with the blank control group,expression levels of IL-5 and eotaxin in peripheral blood were significantly higher (P < 0.01), and the count of EOS and CD34+ cells, as well as CD34+ /CCR3+ significantly increased (P < 0.01) in the model group. Compared with the model group, expression levels of IL-5 and eotaxin, the count of EOS, CD34+ cells, CD34+ / CCR3+ were lowered in three treated groups (P < 0.01). Compared with the Western medicine group, the count of EOS and CD34+ / CCR3+ decreased in the combined group (P < 0.01). The count of EOS was significantly lower in the combined group than in the WD group (P < 0.01).

CONCLUSIONWD could reduce levels of in vivo inflammatory factors, and restrain the differentiation and recruitment of EOS,thereby alleviating the differentiation of CD34 progenitor cells to EOS.

Animals ; Antigens, CD34 ; Asthma, Occupational ; drug therapy ; Bone Marrow ; Cell Differentiation ; Chemokine CCL11 ; Drugs, Chinese Herbal ; therapeutic use ; Eosinophils ; Flow Cytometry ; Interleukin-5 ; Male ; Rats ; Rats, Sprague-Dawley ; Receptors, CCR3 ; Stem Cells

OBJECTIVETo detect the expression level and the mutantion of CD131 in acute myeloid leukemic (AML) cells, and to analyze the relationship of CD131 expression level with clinical features.

METHODSThe peripheral blood mononuclear cells (PBMNC) from 44 AML patients and 25 healthy donors were collected, and the expression level of CD131 mRNA was detected by RT-PCR. The full length coding sequence of CD131 from 15 patients and 5 healthy donors was amplified by RT-PCR, and linked to pGEM-T vector to clone TA, 10 positive recombinant clones of each sample were analyzed by DNA sequencing. The AML patients were divided into CD131 negative and CD131 positive groups according the expression level of CD131, and white blood cell counts, CD34(+) cells percentage, complete remission rate of patients were compared.

RESULTSCD131 was expressed at a lower level in AML than that in healthy donor. Five kinds of CD131 mutantions could be detected in both AML and healthy donor groups, but the mutation rate in AML (75.33%) was higher than that in healthy donors (18.0%). CD131 negative group showed a higher CD34(+) cells percentage (69.1% ± 20.8%), and lower complete remission rate (33.3%) than that in CD131 positive group (69.1% ± 20.8%, 33.3%). No statistically significant difference of WBC counts was found between 2 groups.

CONCLUSIONCD131 is expressed at a low level and shows high frequency of mutation in acute myeloid leukemic cells. CD131 negative AML correlats with high proportion of CD34(+) cells, and showed insensitive to chemotherapy.

Cytokine Receptor Common beta Subunit ; Humans ; Leukemia, Myeloid, Acute ; Leukocyte Count ; Leukocytes, Mononuclear ; Mutation ; RNA, Messenger ; Remission Induction

PURPOSE: Asthma is a pulmonary chronic inflammatory disease characterized by airway obstruction and hyperresponsiveness. Pattern recognition receptors are known to play a key role in the development of allergic diseases as well as host defenses against microbial infection. Receptor interacting protein 2 (RIP2), a serine/threonine kinase, is an adaptor molecule of NOD1 and NOD2, and genetic variation in this receptor is known to be associated with the severity of allergic asthma in children. In this study, we examined the role of RIP2 in the development of allergic airway inflammation in a mouse model. METHODS: Airway inflammation was induced in mice through intranasal administration of ovalbumin (OVA) after 2 intraperitoneal immunizations with OVA. Lung inflammation and mucus hypersecretion were examined histologically and total cell infiltration in bronchoalveolar (BAL) fluids was determined. Levels of the Th2-related cytokines, IL-5 and IL-13, in lung extracts were measured by ELISA. Serum antigen-specific IgE and IgG1 levels were also assessed. RESULTS: OVA-induced lung inflammation and mucus hypersecretion were not different between WT and RIP2-deficient mice. The IL-5 and IL-13 levels in the bronchoalveolar (BAL) fluids were also not impaired in RIP2-deficient mice compared to WT mice. Moreover, RIP2 deficiency did not affect serum OVA-specific IgG1 and IgE levels. CONCLUSIONS: Our results suggest that RIP2 is not associated with the development of allergic airway inflammation.
Administration, Intranasal ; Airway Obstruction ; Animals ; Asthma ; Child ; Cytokines ; Enzyme-Linked Immunosorbent Assay ; Genetic Variation ; Humans ; Immunization ; Immunoglobulin E ; Immunoglobulin G ; Inflammation* ; Interleukin-13 ; Interleukin-5 ; Lung ; Methods ; Mice* ; Mucus ; Ovalbumin ; Ovum ; Phosphotransferases ; Pneumonia ; Receptors, Pattern Recognition

Persistent eosinophil activation in both the upper and lower airway mucosa is a central feature of aspirin-exacerbated respiratory disease (AERD). Eosinophil activation and survival are profoundly influenced by interleukin 5 (IL-5) and its receptor, IL-5R. In patients susceptible to allergic disorders, IL-5 receptor alpha (IL5RA) polymorphisms have been reported; however, an association with AERD remains unclear. We hypothesize that IL5RA polymorphisms may contribute to eosinophil activation in AERD patients. We recruited 139 AERD patients, 171 aspirin-tolerant asthma patients and 160 normal controls. IL5RA polymorphisms (-5993G>A, -5567C>G and -5091G>A) were genotyped and functional activity of polymorphism was assessed by luciferase reporter assay and electrophoretic mobility shift assay (EMSA). There was no significant difference in the genotype frequency of the three polymorphisms among the three groups. AERD patients carrying the AA genotype at -5993G>A had a significantly higher presence of serum-specific immunoglobulin E (IgE) to staphylococcal enterotoxin A (P=0.008) than those with the GG/GA genotype. In vitro, the -5993A allele had a higher promoter activity compared with the -5993G allele in human mast cell (HMC-1; P=0.030) and human promyelocytic leukemia (HL-60; P=0.013) cells. In EMSA, a -5993A probe produced a specific shifted band than the -5993G had. These findings suggest that a functional polymorphism in IL5RA may contribute to eosinophil and mast cell activation along with specific IgE responses to staphylococcal enterotoxin A in AERD patients.
Adult ; Aspirin/*adverse effects ; Electrophoretic Mobility Shift Assay ; Female ; Gene Frequency/genetics ; Humans ; Interleukin-5 Receptor alpha Subunit/*genetics ; Male ; Middle Aged ; Phenotype ; Polymorphism, Single Nucleotide/*genetics ; Respiration Disorders/*chemically induced/*genetics ; Transcription, Genetic

Eosinophils arise from hematopoietic CD34+ stem cells in the bone marrow. They acquire IL-5Ralpha on their surface at a very early stage during eosinophilopoiesis, and differentiate under the strong influence of interleukin (IL)-5. They then exit to the bloodstream, and enter the lung upon exposure to airway inflammatory signals, including eotaxins. In inflamed tissues, eosinophils act as key mediators of terminal effector functions and innate immunity and in linking to adaptive immune responses. Transcription factors GATA-1, CCAAT/enhancer-binding protein, and PU.1 play instructive roles in eosinophil specification from multipotent stem cells through a network of cooperative and antagonistic interactions. Not surprisingly, the interplay of these transcription factors is instrumental in forming the regulatory circuit of expression of eosinophil-specific genes, encoding eosinophil major basic protein and neurotoxin, CC chemokine receptor 3 eotaxin receptor, and IL-5 receptor alpha. Interestingly, a common feature is that the critical cis-acting elements for these transcription factors are clustered in exon 1 and intron 1 of these genes rather than their promoters. Elucidation of the mechanism of eosinophil development and activation may lead to selective elimination of eosinophils in animals and human subjects. Furthermore, availability of a range of genetically modified mice lacking or overproducing eosinophil-specific genes will facilitate evaluation of the roles of eosinophils in the pathogenesis of asthma. This review summarizes eosinophil biology, focusing on development and regulation of eosinophil-specific genes, with a heavy emphasis on the causative link between eosinophils and pathological development of asthma using genetically modified mice as models of asthma.
Aluminum Hydroxide ; Animals ; Asthma ; Biology ; Bone Marrow ; Carbonates ; Eosinophil Major Basic Protein ; Eosinophils ; Exons ; Humans ; Immunity, Innate ; Interleukin-5 ; Interleukins ; Introns ; Lung ; Mice ; Multipotent Stem Cells ; Receptors, CCR3 ; Stem Cells ; Transcription Factors