Identification of Per a 13 as a novel allergen in American cockroach.

BACKGROUND: Cockroaches are an important source of indoor allergens. Environmental exposure to cockroach allergens is closely associated with the development of immunoglobulin E (IgE)-mediated allergic diseases. However, the allergenic components in the American cockroaches are not fully studied yet. In order to develop novel diagnostic and therapeutic strategies for cockroach allergy, it is necessary to comprehensively investigate this undescribed allergen in the American cockroach. METHODS: The full-length cDNA of the potential allergen was isolated from the cDNA library of the American cockroach by PCR cloning. Both the recombinant and natural protein molecules were purified and characterized. The allergenicity was further analyzed by enzyme linked immunosorbent assay, immunoblot, and basophil activation test using sera from cockroach allergic patients. RESULTS: A novel allergen belonging to glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was firstly identified in the American cockroach and named as Per a 13. The cDNA of this allergen is 1255 base pairs in length and contains an open reading frame of 999 base pairs, encoding 332 amino acids. The purified Per a 13 was fully characterized and assessed to react with IgEs from 49.3 % of cockroach allergic patients, and patients with allergic rhinitis were more sensitized to it. Moreover, the allergenicity was further confirmed by immunoblot and basophil activation test. CONCLUSIONS: We firstly identified GAPDH (Per a 13) in the American cockroach, which is a novel type of inhalant allergen derived from animal species. These findings could be useful in developing novel diagnostic and therapeutic strategies for cockroach allergy.

Prevalence of allergen-specific IgE in southern China: a multicenter research.

Identifying allergen distribution is meaningful and significant for effective diagnosis and treatment of allergic diseases. This study compared the allergen sensitivity in four southern China cities. We enrolled 55,432 participants (27,408 male, 28,024 female) between 2007 and 2019. The allergen-specific IgE levels were compared by the χ2 test. The five prevalent sensitivities were for mite mix (10,985, 19.82%), cockroach (4,860, 8.77%), crab (4,450, 8.03%), fish mix (3,874, 6.99%), and house dust (3,486, 6.29%). Almost all allergen sensitivities decreased with age, particularly from infant to middle aged participants (p < 0.05). An exception was Shenzhen, where food allergen positive rates remained constant in all age groups studied. The proportion of male sensitive to at least one food allergen (OR 1.130; 95% CI 1.088-1.174, p < 0.0025) or aeroallergen (OR, 1.117; 95% CI, 1.078-1.158, p < 0.0025) was higher than female in all four cities. Except for dog dander and tree mix, all aeroallergens differed significantly between seasons (p < 0.05). Liuzhou had the highest rates of food allergen- and aeroallergen-positive participants. The allergen-specific IgE distribution differed among the studied cities, with significant seasonal differences. Young age, male sex, and aeroallergens were risk factors for allergic disease.

Respiratory sensitization to insect allergens: Species, components and clinical symptoms.

Airborne insect particles have been identified as an important cause of respiratory allergies, including allergic asthma and rhinitis. In the literature, the significance of respiratory exposure to insect particles as a cause of occupational allergy has been well-documented. Indeed, many cases of occupational allergy have been reported including allergy to the larvae of flies and moths in anglers and occupationally exposed workers, to grain pests in bakers or other workers handling grains, and to crickets and/or locusts in researchers and workers in aquaculture companies. Furthermore, the prevalence of sensitization to insect allergens is considerably high among patients with asthma and/or rhinitis who are not occupationally exposed to insects, suggesting the clinical relevance of exposure to insects in indoor and outdoor environmental non-occupational settings. Exposure to cockroaches, a well-studied indoor insect, is associated with cockroach sensitization and the development and exacerbation of asthma. Booklice, another common indoor insect, were recently identified as a significant sensitizer of asthmatic patients in Japan and India, and potentially of asthma patients living in warm and humid climates around the world. Lip b 1 was identified as an allergenic protein contributing to the species-specific sensitization to booklice. Moths are considered a significant seasonal outdoor allergen and their allergens are considered to have the highest sensitization rate among Japanese patients. However, other than cockroaches, allergenic insect proteins contributing to sensitization have not been fully characterized to date.

CaMKII oxidation regulates cockroach allergen-induced mitophagy in asthma.

BACKGROUND: Autophagy plays an important role in causing inflammatory responses initiated by environmental pollutants and respiratory tract infection. OBJECTIVE: We sought to investigate the role of cockroach allergen-induced excessive activation of autophagy in allergic airway inflammation and its underlying molecular mechanisms. METHODS: Environmental allergen-induced autophagy was investigated in the primary human bronchial epithelial cells (HBECs) and lung tissues of asthmatic mouse model and patients. The role of autophagy in asthma development was examined by using autophagy inhibitor 3-methyladenine in an asthma mouse model. Furthermore, the involvements of reactive oxygen species (ROS) and oxidized Ca2+/calmodulin-dependent protein kinase II (ox-CaMKII) signaling in regulating autophagy during asthma were examined in allergen-treated HBECs and mouse model. RESULTS: Cockroach allergen activated autophagy in HBECs and in the lung tissues from asthmatic patients and mice. Autophagy inhibitor 3-methyladenine significantly attenuated airway hyperresponsiveness, TH2-associated lung inflammation, and ROS generation. Mechanistically, we demonstrated a pathological feedforward circuit between cockroach allergen-induced ROS and autophagy that is mediated through CaMKII oxidation. Furthermore, transgenic mice with ROS-resistant CaMKII MM-VVδ showed attenuation of TH2-associated lung inflammation and autophagy. Mitochondrial ox-CaMKII inhibition induced by adenovirus carrying mitochondrial-targeted inhibitor peptide CaMKIIN suppresses cockroach allergen-induced autophagy, mitochondrial dysfunction, mitophagy, and cytokine production in HBECs. Finally, mitochondrial CaMKII inhibition suppressed the expression of one of the key ubiquitin-binding autophagy receptors, optineurin, and its recruitment to fragmented mitochondria. Optineurin knockdown inhibited cockroach allergy-induced mitophagy. CONCLUSIONS: Our data suggest a previously uncovered axis of allergen-ROS-ox-CaMKII-mitophagy in the development of allergic airway inflammation and asthma.

Shrimp and cockroach co-sensitization in Southern China: Association with moth sensitization.

Background: Moth is a common allergen in southern China. Shrimp sensitization might be related to the moth allergen. Objective: This study investigated sensitization to moth allergen in patients in southern China sensitized to shrimp and explored the effect of moth sensitization on different allergic diseases. Methods: Serum samples from 212 patients sensitized to shrimp were tested for specific immunoglobulin E (sIgE) to Dermatophagoides pteronyssinus, crab, cockroach, and moth. Results: The patients sensitized to shrimp were co-sensitized to D. pteronyssinus (88.7%), crab (85.4%), cockroach (89.2%), and moth (92.0%). Overall, 75% of the patients sensitized to shrimp tested positive to the above allergens; only four patients were sensitized to shrimp alone. The median (interquartile range [IQR]) concentrations of sIgE to shrimp (2.66 kU/L [1.02-6.11 kU/L] versus 1.61 kU/L [0.70-3.67 kU/L]), crab (2.35 kU/L [0.83-4.18 kU/L] versus 1.30 kU/L [0.59-3.14 kU/L]), cockroach (3.78 kU/L [0.98-6.91 kU/L] versus 1.56 kU/L [0.85-3.17 kU/L]), and moth (4.70 kU/L [2.98-9.62 kU/L] versus 2.85 kU/L [1.16-7.01 kU/L]) in patients with skin allergic diseases was significantly higher than in patients with respiratory allergic diseases (all p < 0.05). The median (IQR) concentration of sIgE to cockroach in the young adults (2.33 kU/L [0.86-5.56 kU/L]) was the highest among all age groups as well as to moth (young adults: 4.14 kU/L [1.93-8.24 kU/L]). With the increasing positive class of shrimp allergen, the sIgE concentration of moth, cockroach, and crab also increased, and the optimal scaling analysis showed that the sIgE of crab, cockroach, and moth had a strong correlation with sIgE to shrimp (Cronbach α = 93.8%). Conclusion: This study found a high rate of co-sensitization between moth, D. pteronyssinus, cockroach, and crab among patients sensitized to shrimp and a strong correlation between shrimp, moth, and cockroach. Shrimp and cockroach co-sensitization might be related to moth allergens.

A Novel Serine Protease Inhibitor PE-BBI Ameliorates Cockroach Extract-Mediated Airway Epithelial Barrier Dysfunction.

Epithelial barrier dysfunction, characteristic of allergic airway disease may be, at least in part, due to the action of allergen-associated protease activities. Cockroach allergy is a major global health issue, with cockroaches containing considerable serine trypsin-like protease (TLP) activity. The present study sought to evaluate two novel protease inhibitors (PE-BBI and pLR-HL), recently isolated from amphibian skin secretions, for their potential to neutralise cockroach TLP activity and to determine any protective effect on cockroach-induced airway epithelial barrier disruption. Inhibitor potencies against the cockroach-associated activities were determined using a fluorogenic peptide substrate-based activity assay. 16HBE14o- cells (16HBE; a bronchial epithelial cell line) were treated with cockroach extract (CRE) in the presence or absence of the compounds in order to assess cell viability (RealTime Glo luminescent assay) and epithelial barrier disruption (transepithelial resistance and paracellular dextran flux). PE-BBI potently and selectively inhibited CRE TLP activity (pIC50 -8), but not host (16HBE) cell surface activity, which conferred protection of 16HBE cells from CRE-induced cell damage and barrier disruption. Novel protease inhibitor strategies such as PE-BBI may be useful for the treatment of allergic airway disease caused by cockroach proteases.

Effects of educational efforts in tribal homes and schools to reduce asthma triggers, symptoms and missed school days.

Native American asthma prevalence has been estimated higher than for the U.S. population average, and uncontrolled asthma results in absence from schools. This study analyzed effects of targeted education campaigns in both homes and schools on reducing asthma triggers and symptoms among 119 children with asthma, who were recruited from Cherokee and Nez Perce communities. The education campaigns were developed in collaboration with the researchers and the tribes, tailoring strategies to reduce asthma triggers adapted to tribal lifestyles. There was a special emphasis in identification of Traditional Ecological Knowledge (TEK) aspects, which were woven into the study plan and implementation. Some 62 study homes were investigated and the parents received targeted education at the beginning of the study, whereas the remaining control homes received the same education at the end of the study. In Cherokee homes, allergen levels were significantly increased in both control and study groups throughout the study. In Nez Perce homes, dog and cockroach allergen levels decreased significantly in the study homes. The parents reported asthma control test (ACT) scores of the children were significantly improved in both study and control groups, whereas respiratory illness days were reduced in the study group. In schools, allergen levels, particularly cat, dog and cockroach allergens, increased throughout the year in both study and control schools. However, high contact surface cleaning effectiveness based on adenosine tri phosphate (ATP) readings was improved in the study schools. It was also found that high contact surface cleaning effectiveness may have effects on students' absence rates tracked by schools. In conclusion, the results indicate partial improvements in parent reported health symptoms, although the improvements could not be definitively attributed to reduction of any specific exposure in the home environment. In the school environment, effective cleaning throughout the flu season could result in decreased absence rates.

Association between skin prick test and serum specific immunoglobulin E to American cockroach allergens in allergic rhinitis patients

INTRODUCTION AND OBJECTIVES: American cockroach is a common aeroallergen sensitization in allergic rhinitis (AR) patients. Association between skin prick test (SPT) and specific immunoglobulin E (sIgE) to American cockroach allergen remains uncertain. This study aimed to evaluate the association between SPT and sIgE to American cockroach allergen in patients with AR. MATERIALS AND METHODS: This cross-sectional study was conducted in Thai AR patients aged 6-25 years from September 2013 to October 2014. SPT and sIgE to American cockroach allergen were performed and the correlation was calculated using SPSS Statistics version 18. RESULTS: Sixty-seven AR patients, with median age of 15 years were included in this study. SPT and sIgE to American cockroach allergen showed a positive result in 68.7% and 58.2% cases, respectively. Positive SPT or positive sIgE to American cockroach was found in 79.1%. Thirty-two patients (47.8%) tested positive for both SPT and sIgE to American cockroach allergen. Fourteen from a total of 67 cases (20.9%) with negative sIgE had positive SPT to American cockroach, while seven cases (10.4%) with negative SPT had positive sIgE to American cockroach. Moderate correlation was observed between mean wheal diameter (MWD) and sIgE level to American cockroach (r = 0.465, p = 0.001). No significant correlation was found between MWD of SPT or sIgE level to American cockroach and AR severity. CONCLUSION: A moderate correlation was observed between MWD of SPT and sIgE level to American cockroach. If SPT is negative in allergic rhinitis patients highly suspected of having American cockroach allergy, serum sIgE should be considered and viceversa

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Association between serum immunoglobulin E levels and knee osteoarthritis in Korean adults.

OBJECTIVE: The objective of this study was to examine whether osteoarthritis (OA) in the knees was associated with total immunoglobulin E (IgE), allergen-specific IgE, or allergic sensitizations in a nationally representative population. METHODS: The study population comprised of 785 adults aged 50 years or more in the Korea National Health and Nutrition Examination Survey 2010. OA was diagnosed as radiographic (rOA) and symptomatic osteoarthritis (sxOA). We performed multivariable logistic regression analyses to investigate relationships of OA in a knee with serum total IgE, allergen (Dermatophagoides farinae, cockroach, and dog allergens)-specific IgE, and allergic sensitizations. RESULTS: Participants with the highest tertile of the total IgE had 92% and 242% increased risk of knee rOA and sxOA, respectively. Those with D. farinae-specific IgE had 2.2 times increased risk of knee sxOA compared to the lowest tertile. Participants with high total IgE (>150kU/L) had a 60% increased risk of knee rOA. Those with D. farinae-specific sensitization (>0.35kU/L) had 2.0 times increased risk of knee sxOA in compared to those without sensitization. Population-attributable fractions of knee rOA caused by high total IgE and knee sxOA caused by D. farinae-specific sensitization were 9.8% and 15.3%, respectively. CONCLUSIONS: Total IgE and D. farinae-specific IgE were significantly associated with OA in knees of Korean adults. High total IgE and D. farinae-specific sensitization were also associated with their OA.

Antigenic cross-reactivity between Schistosoma mansoni and allergenic invertebrates putatively due to shared glycanic epitopes.

Previous studies have shown that rabbit IgG antibodies against Schistosoma mansoni egg antigens (SmSEA) cross-react with allergens in natural rubber latex, peanuts and grass and tree pollens. Here we describe antigenic molecules that cross-react with rabbit anti-S. mansoni IgG antibodies in extracts of the house dust mite (HDM) Dermatophagoides farinae, the Australian cockroach (ACR) Periplaneta australasiae and in the venom of the honey bee Apis mellifera (HBV). Tandem mass spectrometry identified the cross-reactive allergens as Der f 15 in HDM, two homologues of the Periplaneta americana cockroach allergen Cr-PI/Per a 3 in ACR and two isoforms of the allergen Api m 1 (phospholipase A2: PLA2) in HBV. Cross-reactive rabbit anti-SmSEA IgG antibodies eluted from the three invertebrate allergens reacted with S. mansoni egg antigens and variably with schistosome cercarial and worm antigens. Treatment of the electroblotted allergens with sodium metaperiodate abrogated most of the cross-reactivity of the rabbit anti-SmSEA antibodies, suggesting it was due to cross-reactive carbohydrate determinants (CCDs). Furthermore, analyses of the allergens' amino acid sequences indicated that they had potential for both N- and O-linked glycosylation. A potential role for the CCDs shared by the schistosome and invertebrates in inducing an allergy-protective effect, as proposed by the hygiene hypothesis, is discussed.

Activation of aryl hydrocarbon receptor (AhR) in mesenchymal stem cells modulates macrophage polarization in asthma.

Macrophage polarization has been demonstrated to exert a vital role on asthma pathogenesis. Mesenchymal stem cells (MSC) have the capacity to modulate macrophage differentiation from a pro-inflammatory M1 phenotype toward an anti-inflammatory M2 phenotype. However, the impact of MSC-macrophage interactions on asthma development and underlying mechanisms responsible for this interaction remain largely unknown. The aim of this study was to investigate the role of AhR expressed on MSC in macrophage polarization in a cockroach extract (CRE)-induced asthma mouse model. The studies here revealed that MSC polarized macrophages from a pro-inflammatory M1 phenotype toward an anti-inflammatory M2 phenotype in this model. The mRNA levels of interleukin (IL)-6, IL-1ß, and NOS2 as M1 markers were significantly decreased while those of select M2 markers such as Arg-1, FIZZ1, and YM-1 were significantly enhanced. It was also observed that aryl hydrocarbon receptor (AhR) signaling was significantly increased during asthma pathogenesis as demonstrated by enhanced mRNA expression of AhR, CYP1a1, and CYP1b1. It was also seen that the elevated AhR signaling was able to attenuate the onset of asthma. Use of an AhR antagonist (CH223191) resulted in significant inhibition of the AhR signaling and increases in M2 marker expression, but led to elevation of expression of M1 markers in the CRE-induced asthma model. Taken together, the current study showed that MSC can modulate macrophage polarization, in part, via activation of AhR signaling during CRE-induced asthma.

Cockroaches: Allergens, Component-Resolved Diagnosis (CRD) and Component-Resolved Immunotherapy.

Allergic diseases are assuming increasing trend of prevalence worldwide. The diseases confer increasing demand on medical and healthcare facilities. Patients with allergies have poor quality of life and impaired cognition. Adult patients have subpar working efficiency while afflicted children are less effective at school, often have school absenteeism and need more attention of their caregivers. All of them lead to negative socio-economic impact. This narrative review focuses on cockroach allergy including currently recognized cockroach allergens, pathogenic mechanisms of allergy, componentresolved diagnosis and allergen-specific immunotherapy, particularly the component-resolved immunotherapy and the molecular mechanisms that bring about resolution of the chronic airway inflammation.

The level of allergens in dust samples collected from selected schools in Shiraz, Iran and its asthma-risk implications.

BACKGROUND: Both home and school are important places where children are exposed to various indoor allergens. This study aimed to identify the profile of indoor allergens in schools and its impact on asthma development. METHODS: A total of 104 classrooms from 52 schools were selected for dust collection during the fall of 2017. The levels of indoor allergens including dust mite (Der f1, Der p1), cat (Fel d1), cockroach (Bla g1) and mouse (Mus m1) were measured by enzyme linked immunosorbent assay (ELISA). The diagnosis of asthma was made in all students of the selected classes by the allergist. The collected data were analyzed using SPSS version 21.0. RESULTS: Out of 2816 students in the selected classes, 180 students were involved with asthma. Students were mostly exposed to Bla g1 (83.1%), followed by Der f1 (51.5%), Mus m 1 (45.5%), Der p1 (8.9%) and Fel d1 (7.9%) in the dust collected from 101 classrooms. Although levels of all studied allergens in the settled dust of the classrooms were low, there was a relationship between Fel d1 in the classroom dust and development of asthma. CONCLUSION: This study showed considerable levels of cockroach allergens in schools. Exposure to cat allergen in our schools played an important role in asthma development; further school-based investigations require evaluating the role of classroom allergen on asthma development.

Avoidance of allergens as an environmental method in the prevention of inhaled allergy symptoms.

Asthma and other inhaled allergies are some of the most common paediatric diseases. The association of exposure to allergens with induction and exacerbation of symptoms has been proven. The majority of allergens are permanently or periodically suspended in the air, which leads to impaired quality of life for sensitive patients. Therefore, many methods of prevention and therapy of allergic diseases have been developed. The method of allergen exposure avoidance is often the first and the most significant measure. The present research has been conducted to evaluate, based on scientific data, which measures have the most reliable evidence of effectiveness. Environmental allergen avoidance methods, despite limited evidence supporting their clinical efficacy, are listed as the main therapeutic approaches in most recommendations. The significance of the holistic approach is also emphasised: only simultaneous introduction of several avoidance methods can bring possibly beneficial effects for the patient.

Optimal conditions for the storage of German cockroach extract.

Allergen extracts are commonly utilized for diagnosis and immunotherapy; however, the stability of protease­rich extracts is important for a precise diagnosis and treatment efficacy. The present study determines the optimal conditions for the storage of German cockroach allergen extract. Cockroach extracts were reconstituted in four buffers: normal saline (NS), 50% glycerol in NS, 0.3% phenol in NS, or 0.3% phenol and 50% glycerol in NS. The extracts in different buffers were stored either at room temperature (18­26˚C, RT) or refrigerated (2­8˚C). Subsequently, the protein concentration and allergen content (Bla g 1 and Bla g 2) in the extracts were examined for the course of one year. Extract potency was estimated by inhibition ELISA. At least 90.5% protein, 94.4% Bla g 1, 65.2% Bla g 2, and 91.4% potency remained after one year when 50% glycerol NS was added to the extract with refrigeration. However, less than 13.7% protein, 17.1% Bla g 1, 0% Bla g 2 and 32.5% potency were maintained after one year when 50% glycerol NS was not added to the extract and was maintained at RT. The addition of 0.3% phenol NS did not show significant effects on extract stability. The addition of 50% glycerol NS and refrigerated storage temperature were found to be important factors for increasing the shelf life of protease­rich cockroach extract.

Identifying Potential Co-Sensitization and Cross-Reactivity Patterns Based on Component-Resolved Diagnosis.

BACKGROUND: Component-Resolved diagnosis (CRD) can help to establish immunoglobulin E (IgE) sensitization profiles and potential risks and determines whether specific IgE is the result of primary sensitization or cross-reactivity, especially for those who are polysensitized. METHODS: We recruited 432 patients with mite-sensitized respiratory allergic diseases to study the co-sensitization and cross-reactivity of the 17 allergen components in Guangdong Province, China, using the CRD method and to describe the potential association between allergen components. RESULTS: Among the 432 patients, serum specific immunoglobulin E of the 17 components were tested by EUROIMMUN system. Der p 1 (81.48%), Der f 2 (77.78%), Der f 1 (74.07%), Der p 2 (66.20%) and Der p 23 (54.63%) were the main sensitized components in patients with mite-sensitized respiratory allergy, while the components of cockroach, crab, and shrimp had a lower positive rate. In the crude extract allergen-positive samples, Der f 2 (91.06%) and Der f 1 (86.72%) were the major sensitized components of Der f, while Der p 1 (94.52%), Der p 2 (78.36%), Der p 23 (63.29%) were the major sensitized components of Der p, And other components of Der p such as Der p 7 (34.25%), Der p 5 (17.81%), Der p 10 (12.05%), Der p 3 (1.92%) were all below 50.00%. Blo t 5 (54.55%) was one of the major components of Blo t. The positive rates of all Bla g components were as follows, rBla g 2 (15.56%) >rBla g 5 (8.89%) >rBla g 4 (4.44%) >rBla g 1 (1.11%). The positive rate of the only available pen a 1 component was 9.43%. Using hierarchical cluster and optimal scale analysis, 17 components can be roughly divided into 5 different sensitization clusters. Also, from the results of the Venn diagram, the allergen component in each cluster has a high proportion of co-sensitization and cross-reactivity. Regardless of age, total IgE levels, and disease type factors, similar sensitization profiles were observed for each component in the same category based on hierarchical clustering analysis. CONCLUSIONS: Epidemiological data on allergen components causing allergic symptoms can be further understood using CRD. Der p 1, Der p 2, Der p 23, Der f 1, Der f 2 as the primary sensitizing component of the study cohort. The positive rate for Blo t 5 was 28.01% for all populations and 54.45% for Blo t-positive samples. In addition, CRD allows us to identify more potential allergen associations such as common sensitivities and cross-reactions between component proteins. Based on these results, we suggest that when patients are identified as sensitized to a particular allergen, clinicians can pay more attention to other allergy components that are closely related to it.

Inhibition of Bruton's tyrosine kinase and IL-2 inducible T-cell kinase suppresses both neutrophilic and eosinophilic airway inflammation in a cockroach allergen extract-induced mixed granulocytic mouse model of asthma using preventative and therapeutic strategy.

Asthma is a complex airways disease with a wide spectrum which ranges from eosinophilic (Th2 driven) to mixed granulocytic (Th2/Th17 driven) phenotypes. Mixed granulocytic asthma is a cause of concern as corticosteroids often fail to control this phenotype. Different kinases such as Brutons's tyrosine kinase (BTK) and IL-2 inducible T cell kinase (ITK) play a pivotal role in shaping allergic airway inflammation. Ibrutinib is primarily a BTK inhibitor, however it is reported to be an ITK inhibitor as well. In this study, we sought to determine the effect of Ibrutinib on Th1, Th17 and Th2 immune responses in a cockroach allergen extract (CE)-induced mixed granulocytic (eosinophilic and neutrophilic) mouse model in preventative mode. Ibrutinib attenuated neutrophilic inflammation at a much lower doses (25-75 µg/mouse) in CE-induced mixed granulocytic asthma whereas Th2/Th17 immune responses remained unaffected at these doses. However, at a much higher dose, i.e. 250 µg/mouse, Ibrutinib remarkably suppressed both Th17/Th2 and lymphocytic/neutrophilic/eosinophilic airway inflammation. At molecular level, Ibrutinib suppressed phosphorylation of BTK in neutrophils at lower doses and ITK in CD4 + T cells at higher doses in CE-treated mice. Further, effects of Ibrutinib were compared with dexamethasone on CE-induced mixed granulocytic asthma in therapeutic mode. Ibrutinib was able to control granulocytic inflammation along with Th2/Th17 immune response in therapeutic mode whereas dexamethasone limited only Th2/eosinophilic inflammation. Thus, Ibrutinib has the potential to suppress both Th17/Th2 and neutrophilic/eosinophilic inflammation during mixed granulocytic asthma and therefore may be pursued as alternative therapeutic option in difficult-to-treat asthma which is resistant to corticosteroids.