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Kryvko Yurij, Soguiko Yurij, Hot Iryna, Antonenko Yuliia, Baida Mariana, Kovalska Marta. Peculiarities of changes in a functional condition of pro-oxidant and antioxidant systems in guinea pigs’ lungs in experimental allergic alveolitis under the conditions of immobilization stress. Journal of Education, Health and Sport. 2021;11(09):798-805. eISSN 2391-8306. DOI http://dx.doi.org/10.12775/JEHS.2021.11.09.095 https://apcz.umk.pl/JEHS/article/view/JEHS.2021.11.09.095 https://zenodo.org/record/5554986 The journal has had 5 points in Ministry of Science and Higher Education parametric evaluation. § 8. 2) and § 12. 1. 2) 22.02.2019. © The Authors 2021; This article is published with open access at Licensee Open Journal Systems of Nicolaus Copernicus University in Torun, Poland Open Access. This article is distributed under the terms of the Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author (s) and source are credited. This is an open access article licensed under the terms of the Creative Commons Attribution Non commercial license Share alike. (http://creativecommons.org/licenses/by-nc-sa/4.0/) which permits unrestricted, non commercial use, distribution and reproduction in any medium, provided the work is properly cited. The authors declare that there is no conflict of interests regarding the publication of this paper. Received: 25.08.2021. Revised: 12.09.2021. Accepted: 30.09.2021. UDC: 616.24-002-008.6-056.3-092: (612.24.015.11+612.014.484)]-092.9 peculiarities of changes in a functional condition of pro-oxidant and antioxidant systems in guinea pigs’ lungs in experimental allergic alveolitis under the conditions of immobilization stress Yurij Kryvko1, Yurij Soguiko1, Iryna Hot¹, Yuliia Antonenko¹, Mariana Baida, Marta Kovalska Danylo Halytsky Lviv National Medical University, Lviv 1Andrei Krupynskyi Lviv Medical Academy, Lviv dr.kowalska.marta@gmail.com Abstract We have analyzed the results of investigation of alterations in indices of pro-oxidant (conjugated diene and malondialdehyde) and antioxidant (superoxide dismutase, ceruloplasmin, catalase) systems in guinea pigs’ lungs in experimental allergic alveolitis in the dynamics of EAA development under the conditions of immobilization stress. The investigation was conducted on 62 female guinea pigs weighing 180-220 g, divided into 4 groups: I – intact guinea pigs (n=20), II – guinea pigs (n=14) with EAA under the conditions of IS (1st day); III – guinea pigs (n=14) with EAA under the conditions of IS (2nd day); IV – guinea pigs (n=14) with EAA under the conditions of IS (64th day). The results of experimental investigation showed that a significant increase in conjugated diene level in animals’ lungs was observed at all stages of EAA development under the conditions of immobilization stress as compared with control group, indicating activation of this marker. The same changes occurred with MDA content, indicating excessive accumulation of this lipid peroxidation product in lung tissue. Intensive synthesis of free radical compounds caused activation of some components of enzymatic system of antioxidant defense. In particular, a moderate decrease in superoxide dismutase activity in lung tissue is observed in response to increased level of LOPs at early stages of EAA and immobilization stress development as compared with the indices in intact animals. The same situation is observed with catalase and ceruloplasmin activity in the lungs of guinea pigs with modeled AA and IS. Key words: experimental allergic alveolitis; immobilization stress; peroxide lipid oxidation; antioxidant system.

Context Modified BuShenYiQi formula (M-BYF) is derived from BuShenYiQi formula, used for the treatment of allergic asthma. The exact effect and mechanism of M-BYF on the improvement of asthma remain unclear. Objective We investigated the mechanism underlying the therapeutic effect of M-BYF on allergic asthma. Materials and methods The asthma model was established in female BALB/c mice that were sensitized and challenged with ovalbumin (OVA). Mice in the treated groups were orally treated once a day with M-BYF (7, 14 and 28 g/kg/d) or dexamethasone before OVA challenge. Control and Model group received saline. Pathophysiological abnormalities and percentages of lung type 2 innate lymphoid cells (ILC2s) and Th9 cells were measured. Expression levels of type 2 cytokines and transcription factors required for these cells function and differentiation were analysed. Expression of vasoactive intestinal polypeptide (VIP)–VPAC2 signalling pathway-related proteins, and percentages of VIP expressing (VIP+) cells and VPAC2, CD90 co-expressing (VPAC2+CD90+) cells were detected. Results M-BYF alleviated airway hyperresponsiveness, inflammation, mucus hypersecretion and collagen deposition in asthmatic mice. M-BYF down-regulated percentages of ILC2s and Th9 cells with lower expression of GATA3, PU.1 and IRF4, reduced IL-5, IL-13, IL-9 and VIP production. The decrease in the expression of VIP–VPAC2 signalling pathway and percentages of VIP+ cells, VPAC2+CD90+ cells were observed after M-BYF treatment. The LD50 value of M-BYF was higher than 90 g/kg. Discussion and conclusions M-BYF alleviated experimental asthma by negatively regulating ILC2s and Th9 cells and the VIP–VPAC2 signalling pathway. These findings provide the theoretical basis for future research of M-BYF in asthma patient population.

Background: Multiple sclerosis (MS), an inflammatory disease of the central nervous system (CNS), leads to demyelination, neuronal injury, and loss of white matter, yet still can't be cured. Exosomes are double-layered membrane vesicles of 30–200 nm in size, which can easily penetrate the blood–brain barrier (BBB). Exosomes derived from umbilical cord mesenchymal stem cells exosomes (UMSC-exos) has been shown to treat experimental autoimmune encephalomyelitis (EAE) through the action of anti-inflammatory and immunomodulatory, but its clinical translation has been hampered by their inefficacious accumulation in CNS. Therefore, we developed a TAxI-peptide-chimeric UMSC-exos termed TAxI-exos for CNS-specific accumulation and curative effect in EAE.Methods: We used an EAE model in vivo, and actived T cells and BV-2 cells models in vitro. After two immunizations to establish the EAE model, UMSC-exos, TAxI-exos or DiR labeled exosomes were administered to EAE mice EAE mice for one dose (150μg) before the peak at day 15. On day 30, the mice were sacrificed to collect spinal cords, spleens, and blood for analysis of demyelination, inflammation, microglia, the proportions of T-cell subsets, and the expression of inflammatory cytokines. In vitro, for immune mechanism analyses, PBMCs and splenocytes isolated from healthy C57BL/6 mice, were activated and incubated with 0.15mg/mL UMSC-exos or TAxI-exos. Activated BV-2 cells were used to explore the targeting-ability and polarization-regulating ability of UMSC-exos and TAxI-exos.Results: As expected, TAxI-exos had significant curative effects in EAE mice compared with UMSC-exos via an enhanced targeting-ability. The treatment alleviated inflammation, facilitated microglial cell polarization from M1 to M2, reduced the proportions of T-cell subsets, increased the expression levels of IL-4, IL-10, TGF-β, and IDO-1, and decreased the levels of IL-2, IL-6, IL-17A, IFN-γ, and TNF-α. Conclusions: TAxI-exos have a great CNS-targeting ability and suppress pathological processes in EAE mice, which have a great potential therapeutic utility for MS and other CNS diseases.

Objective: Allergic rhinitis (AR) is an inflammatory disease of the nasal mucosa mediated by IgE after exposure to an allergen. The most well known related comorbidity of AR is asthma. This study was planned due to the need for an animal model for studies on AR-asthma coexistence. In this study, the frequency of AR accompanying in the asthma model created in mice,and the usability of the related model in AR studies will be investigated. Methods: In our study, 6-8 week-old, 18-20 g BALB/c mice were used. Chicken egg ovalbumin (OVA Grade V, Sigma) was administered through intraperitoneal (IP) route at doses of 10 μg on days 0 and 14. Mice were exposed to aerosolized 2.5% ovalbumin solution in sterile saline for 30 minutes 3 days a week for 8 weeks, starting 7 days after the last IP administration (21st day). After exposure to OVA, mice were observed for typical signs of AR including sneezing, runny nose, and nasal itching. The final diagnosis of AR was made by histopathological examination of the rhinotracheal tissues of mice. Results: In our study, all mice exposed to ovalbumin received histopathologic diagnosis of AR. Increased number of capillaries lymphocytes, polymorphonuclear leukocytes and eosinophilsper square millimetre of rhinotracheal tissues were calculated in the murine model of AR compared to the the control group. Conclusion: This study introduced a new AR model, not cited in the literature, and induced with the longest-term ovalbumin exposure in the literature. It was concluded that this model, known as the asthma model, can also be used to induce an AR model and can be used in studies investigating coexistence of allergic rhinitis and asthma.

Purpose To investigate the effect of biospray dressing on the extent of eosinophil infiltration in the nasal mucosa and the level of serum IgE in experimental allergic rhinitis with nasal provocation. Method Twenty-four BALB/c mice were randomly divided into the normal control group, allergic rhinitis (AR) group, dexamethasone (DEX) treatment group, and biospray dressing (BD) group. The mice in the latter 3 groups were prepared for animal models of AR according to standard protocols. Mice in the BD group were administered a nasal spray before the nasal provocation, and those in the DEX group were administered an intraperitoneal injection of DEX. The nasal mucosa and serum were collected from each group. Nasal mucosa eosinophil infiltration was evaluated using hematoxylin and eosin staining, and enzyme-linked immunosorbent assay (ELISA) was performed to analyze the serum IgE expression. Results Eosinophil infiltration (AR vs BD P = .009) in the nasal mucosa and serum IgE expression (AR vs BD P = .001) were significantly lower in the BD group than in the AR group. There were no significant differences in the extent of eosinophil infiltration in the nasal mucosa or serum IgE expression between the BD and DEX groups. Conclusion Biospray dressings can significantly reduce allergen provocation in the nasal cavity and have a protective effect on the nasal mucosa. They can be used for the prevention and treatment of AR.

International audience; Intravenous immunoglobuin (IVIG) exerts protective effects in experimental allergic bronchopulmonary aspergillosis (ABPA) via a sialylation-dependent mechanism. The protection was associated with reduced recruitment of eosinophils, diminished goblet cell hyperplasia, suppressed Th2 and Th17 responses and reciprocally enhanced regulatory T cells and IL-10, and decreased IgE levels in the circulation.

We investigate whether lycopene has a protective effect in an experimental rat model of allergic rhinitis.Experimental animals (65 rats) were randomized to 7 groups (Sham-Control, Lycopene 10 mg/kg/day, Lycopene 20 mg/kg/day, Intranasal lycopene drops, Intranasal steroid, Corn oil, Allergic Rhinitis). Rats were sensitized by administering of ovalbumin intraperitoneally and intranasally. In addition to ovalbumin; lycopene, corn oil and steroids were given to the relevant groups. Nasal symptom scores of the rats were recorded throughout the study. At end of the study, after intracardiac blood sample collection, all rats were sacrificed, and nasal tissues were examined histopathologically. Serum total immunoglobulin E (IgE) and ovalbumin (OVA) specific IgE were studied from all rats before and after the study.There was a statistically significant increase (p 0.05) in OVA specific IgE values measured before and after the study in all groups except the sham group. In serum total IgE values; there was a statistically significant increase after treatment in allergic rhinitis, corn oil, lycopene 10 mg and intranasal lycopene drops group, but other groups did not show any significant change. Histopathological study with hematoxylin-eosin staining and cyclooxygenase-2 (COX-2), matrix metalloproteinase-9 (MMP-9), vasoactive intestinal peptide (VIP) expression found that lycopene suppresses inflammation with both nasal administration and increased dose. Nasal symptom scores were observed to decrease significantly in all lycopene and steroid groups compared to allergic rihinits and corn groups.It was determined that lycopene were effective in the treatment of allergic rhinitis, and this effect was found to be stronger with increasing doses of lycopene.

Objective. To study the efficacy of release-active forms of antibody to S100 protein in an animal model of multiple sclerosis. Materials and methods. The study was performed on 60 female rats aged 12 weeks. Experimental allergic encephalomyelitis was induced by administration of spinal cord homogenate at the base of the tail. Female rats then received intragastric solution of release-active antibodies to S100 protein (Tenoten) at a dose of 2.5 ml/kg/day or distilled water for 30 days. Positive controls received i.m. injections of glatiramer acetate at a dose of 4 mg/kg/day (Copaxone). Results and discussion. Administration of release-active forms of antibody to S100 increased the latent period of disease onset, decreased in peak disease intensity, and compensated for weight loss in the animals, as compared with controls. The effect of the agents was comparable with that of the Copaxone group.

Aim: Allergic rhinitis (AR) is a common disease. The therapeutic efficacy of AR needs to be improved. This study aims to evaluate the effects of local administration of probiotic extracts on inhibiting experimental AR. Methods: Epithelial cells (ECs) were primed by exposing to Clostridium butyricum extracts (CBe) in the culture to upregulate the expression of IL-10. A mouse AR model was developed to assess the therapeutic potential of CBe in AR. Results: CBe markedly induced the expression of IL-10 in ECs. Co-culture of naive B cells with CBe-primed ECs significantly increased IL-10 expression in the B cells (iB10 cells). The iB10 cells showed immune suppressive function in suppressing effector CD4+ T-cell proliferation. Treatment with nasal drops containing CBe efficiently inhibited experimental AR in mice. Conclusion: Local administration of CBe can efficiently inhibit experimental AR.

Multiple sclerosis (MS) is a chronic disease of the CNS affecting people of working age, in which myelin and the cells producing it, and also neurons, become the targets for aggressive immune cells. It has been suggested that common childhood infections in later childhood increase the risk of developing MS. We report here studies of the course of experimental allergic encephalomyelitis (EAE) in rats given interleukin-1β (IL-1β) at different stages of early postnatal ontogeny. EAE was induced in rats at age three months by single subcutaneous immunizations with homologous spinal cord homogenate in complete Freund’s adjuvant. The number of sick animals was recorded daily, as were the severity and duration of disease. EAE was found to have a more severe course after administration of IL-1β in weeks 1 and 4 of life than in rats of the corresponding control groups. The harmful or protective consequences of IL-1β administration at different periods of early postnatal ontogeny are discussed, as are the role of stress reactivity and its link with the “hygiene hypothesis.”

The recent studies have showed neural stem cells (NSCs) transplantation promoted repair and functional recovery of chronic experimental allergic encephalomyelitis (EAE) animal model for MS based on the concept of cell replacement. In the present study, NSC...

Introduction A major clinical challenge in treating allergic contact dermatitis (ACD) is that the first line of treatment is based on the use of corticosteroids. In this study, we aimed to develop a formulation that is devoid of steroids. Methods We used mouse ears treated with dinitrofluorobenzene (DNFB) to induce ACD. The efficacy of the test formulation to ameliorate and to prevent induced ACD was determined. Results To treat this experimentally induced ACD, we developed a formulation containing BIPxine (a mixture of Rosa moschata and Croton lechleri (antioxidants) and Aloe vera and d-panthenol (moisturizers), and hydroglycolic solutions of disodium cromoglycate. Our results show that clear inhibition of ACD took place. The target of this formulation was PAR-2, TRPV4, and other mediators of the inflammatory and pain responses. However, this formulation must be evaluated in other models besides the mouse to confirm its effectiveness. Conclusion The formulation presented here may provide new ACD therapies that do not involve the use of corticosteroids. Electronic supplementary material The online version of this article (10.1007/s13555-018-0223-8) contains supplementary material, which is available to authorized users.

On the model of experimental analogue of human multiple sclerosis we studied the effects of the mesenchymal stem cells transfected with plasmid vector containing gene IL-10 (MSCs-T) on the functional state of the CNS in rats.Materials and methods. The experimental allergic encephalomyelitis (EАЕ) was induced with spinal cord homogenate of rats with complete Freund’s adjuvant. The mesenchymal stem cells (MSCs) were isolated by the explants technique from Wharton’s jelly of the human umbilical cord and culture up to two passages. Then the MSCs of second passage were transfected of plasmid vector with gene IL-10 and marker gene of green fluorescent protein. Cell transplantation was performed suboccipitally on the 17th day at a dose of 1 million cells in 100 µl of saline per animal.Results. In the open field test we have established that the use of MSCs-T transfected with gene IL-10 suppressed the vertical locomotor activity and elevated the emotional activity as well as partially corrects horizontal locomotor activity indexes which approach the indexes of intact animals.Conclusions. The use of MSCs transfected with plasmid vector with gene IL-10 in the rats with induced EAE is more effective method than treatment using non-transfected MSCs. Combined treatment with IL-10 + MSCs in ЕАЕ rats is more effective than treatment with transfected МSCs.

On the rat model of experimental allergic encephalomyelitis (EAE), as analogue of multiple sclerosis of human, we studied the effect of Wharton’s jelly-derived mesenchymal stem cells and interleukin-10 on the functional parameters of the CNS.Materials and methods. EAE was induced with spinal cord homogenate of rats with complete Freund’s adjuvant. MSCs were isolated by the explants technique from Wharton’s jelly of the human umbilical cord and cultured up to two passages. Recombinant IL-10 was administered intravenously on the day 10 after induction of EAE and subocipitally on the day 17 at a dose of 0.2 mg per animal. Cell transplantation performed subocipitally on the day 17 at a dose of 106 cells per animal. Behavioral reactions studied in the “open field” test three times: on the day 12, 15 and 24 after the induction of EAE.Results. Induced EAE results in significant changes in adaptive behavior of rats on the 12th day as suppression of orienting-exploratory and activation of emotional activity. The application of MSCs and IL-10 has normalized the rates of cognitive activities (orienting-exploratory activity) and emotional sphere (the level of fear, anxiety).Conclusion. Combined treatment of induced EAE in rats with interleukin-10 and human Wharton’s jelly-derived mesenchymal stem cells is effective at correction of behavioral responses of animals.

This chapter focuses on the basic elements of computational drug repositioning and overviews the various in silico, in vivo, and in vitro approaches. It describes the use of Systematic Drug Repositioning (SDR) technologies to predict hitherto unknown adverse drug reactions (ADRs), or to derive mechanistic explanations of known ADRs. The chapter discusses the intellectual property (IP) and commercial aspects of SDR, which are among the major factors that will determine its success. It shows that Pirlindol is neuroprotective in MOG-induced experimental allergic encephalomyelitis, a mouse model of progressive multiple sclerosis (MS). The chapter identifies the risk factors that may increase an individual's susceptibility to this ADR. It illustrates a process through which computational drug repositioning methods can suggest mechanistic hypotheses that can help explain an ADR and propose modifiable risk factors. While computational SDR uses the mechanism of action of the drugs under study to identify novel indications, in vitro/in vivo screening uses the opposite approach.

Objective(s): Experimental allergic encephalomyelitis (EAE) is an autoimmune disease validated as animal model of multiple sclerosis (MS). Administration of genistein, a phytoestrogenic component of soy, to mice at the onset of EAE is known to attenuate the clinical signs of the disease. The potential effects of genistein on established EAE is less studied. In the current study, we aimed to compare the effects of genistein administration on EAE severity in early and late phases of the disease. Materials and Methods: The C57BL/6 mice were induced with EAE, using MOG 35-55 and gavaged with genistein (300 mg/kg) either after the appearance of the first clinical sign or 30 days post disease induction for ten days. 24 hr after the last gavage, mice were sacrificed. Brains and spleens were removed for assessing lymphocyte proliferation, cell cytotoxicity, and cytokine profile. Spinal cords were dissected to assess the amount of demyelination using Luxol fast blue/cresyl violet staining. Results: Administering mice with genistein, after the establishment of EAE, did not reverse the clinical signs of disease. However, treating with genistein at the onset of disease alleviated the clinical signs by reducing neuronal demyelination. Genistein suppressed the production of IFN-γ and enhanced IL-10 secretion in splenocyte and brain. Genistein also reduced IL-12 and TNF-α secretion in splenocytes, suppressed the proliferation of T-cells, and reduced the cell cytotoxicity. Conclusion: Genistein oral therapy might only reduce EAE severity if started in early phases of the disease.

Inflammation modulates tissue damage in relapsing-remitting multiple sclerosis (MS) both acutely and chronically, but its severity is difficult to evaluate with conventional MRI analysis. In mice with experimental allergic encephalomyelitis (EAE, a model of MS), we administered ultra small particles of iron oxide to track macrophage-mediated inflammation during the onset (relapse) and recovery (remission) of disease activity using high field MRI. We performed MRI texture analysis, a sensitive measure of tissue regularity, and T2 assessment both in EAE lesions and the control tissue, and measured spinal cord volume. We found that inflammation was 3 times more remarkable at onset than at recovery of EAE in histology yet demyelination appeared similar across animals and disease course. In MRI, lesion texture was more heterogeneous; T2 was lower; and spinal cord volume was greater in EAE than in controls, but only MRI texture was worse at relapse than at remission of EAE. Moreover, MRI texture correlated with spinal cord volume and tended to correlate with the extent of disability in EAE. While subject to further confirmation, our findings may suggest the sensitivity of MRI texture analysis for accessing inflammation.

Developing a new and effective therapeutic approach against multiple sclerosis (MS) is always an important area of research. RNS60 is a bioactive aqueous solution generated by subjecting normal saline to Taylor-Couette-Poiseuille flow under elevated oxygen pressure. Recently we have demonstrated that RNS60, administered through intraperitoneal injection, ameliorated clinical symptoms and disease progression of experimental allergic encephalomyelitis (EAE), an animal model of MS. Since the intravenous route is not preferred for treating a chronic condition, we tested if nebulization of RNS60 could attenuate the disease process of adoptively-transferred EAE in mice. Although we could not directly image RNS60 after nebulization, nebulized Alexa680 reached spleen, spinal cord and different parts of the brain. Nebulization of RNS60 starting from the acute phase attenuated clinical symptoms of relapsing-remitting EAE in female SJL/J mice. RNS60 nebulization also inhibited perivascular cuffing, maintained the integrity of blood-brain and blood-spinal cord barriers, suppressed inflammation, normalized the expression of myelin genes, and blocked demyelination in the CNS of EAE mice. On the immunomodulatory front, nebulization of RNS60 to EAE mice led to the enrichment of anti-autoimmune regulatory T cells (Tregs) and suppression of autoimmune Th17 cells. Together, these results suggest that nebulization of RNS60 may be used to control aberrant immune responses in MS and other autoimmune disorders.

Experimental allergic alveolitis (AA) in conditions of adrenal myocardial damage (AMD) before treatment was accompanied by an increase in blood levels of phagocytic number and phagocytic index. Application of thiotriazoline resulted in a decrease in phagocytic activity of leukocytes in the blood in AA and AMD.

We obtained data that substantiate the possibility of using the level of sensibilization of immunocompetent cells as an assessment criterion of the severity of experimental allergic encephalomyelitis (EAE), as well as to determine the therapeutic efficacy of using cryopreserved fetal nerve cells (cFNCs) for treatment of this pathology. We demonstrated the dependence of a drop in the autoreactivity of lymphocytes in rats with EAE on the type of injected cellular material cryopreserved in different regimes. We discuss possible mechanisms underlying realization of the therapeutic effect of cFNAs.

This study investigated the antispasticity potential of Sativex in mice. Chronic relapsing experimental allergic encephalomyelitis was induced in adult ABH mice resulting in hind limb spasticity development. Vehicle, Sativex, and baclofen (as a positive control) were injected intravenously and the “stiffness” of limbs assessed by the resistance force against hind limb flexion. Vehicle alone caused no significant change in spasticity. Baclofen (5 mg/kg) induced approximately a 40% peak reduction in spasticity. Sativex dose dependently reduced spasticity; 5 mg/kg THC + 5 mg/kg CBD induced approximately a 20% peak reduction; 10 mg/kg THC + 10 mg/kg CBD produced approximately a 40% peak reduction in spasticity. Sativex has the potential to reduce spasticity in an experimental mouse model of multiple sclerosis (MS). Baclofen reduced spasticity and served as a positive control. Sativex (10 mg/kg) was just as effective as baclofen, providing supportive evidence for Sativex use in the treatment of spasticity in MS.

Calcium signaling is essential for all the functions of T lymphocytes, including those of Th2 cells. Th2 lymphocytes producing interleukins 4, 5 and 13 orchestrate allergic diseases including asthma. T-cell activation induces an influx of Ca(2+) from the external medium through ORAI calcium channels although other calcium channels are likely to be involved. Among them, voltage-gated calcium (Ca(v)1) channels have been reported in some T-cell subsets including Th2 cells. The inhibition of Ca(v)1 channels abrogates T-cell receptor-driven calcium influx and interleukin production by Th2 cells. From a therapeutic point of view, the inhibition of Ca(v)1 channels prevents Th2-dependent experimental allergic asthma. In this review, we will discuss the singularities of calcium responses depending upon the T-cell subset and its state of activation.

Background: Recent studies have shown that transforming growth factor-β1 (TGF-β1) plays an important role in the progression of allergic diseases. Methods: Mouse models of allergic rhinitis were established by ovalbumin sensitization and challenge. Immunostaining was used to analyze the expression of TGF-β1 in the mouse nasal mucosa. A chemotaxis assay was conducted to analyze the impact of TGF-β1 stimulation on migration of mast cells differentiated from mouse bone marrow cells. Chemotaxis and Western blot analysis were further applied to investigate the pathways involved in mast cell migration induced by TGF-β1 stimulation. Results: TGF-β1 expression was induced in allergic rhinitis and phosphorylated Smad2 was expressed in mast cells present in the nasal mucosa. TGF-β1 could induce migration of mast cells, but HTS466284, a TGF-β receptor 1 kinase inhibitor, inhibited this chemotactic activity. After TGF-β1 stimulation, mast cell RhoA expression was significantly increased. TGF-β1-induced mast cell chemotaxis could be inhibited by the RhoA inhibitor Tat-C3 and myosin light chain kinase inhibitor ML-7. Conclusion: TGF-β1 plays a major role in inducing the accumulation of mast cells in allergic rhinitis.

We evaluated the use of immunological biomarkers in transconjunctival immunotherapy by using cholera toxin B for treating experimental allergic conjunctivitis in a mouse model.Balb/c mice were sensitized using intraperitoneal injections of ovalbumin (OVA) and were then divided into two groups. The first group was treated by topical instillation of OVA after the instillation of combined OVA and cholera toxin B (CTB) solution B group). The second group was treated by topical instillation of OVA alone (allergy group). The control group consisted of nonsensitized mice undergoing topical OVA instillation only. The numbers of eosinophils and CD4-positive lymphocytes in the conjunctiva were determined histologically, and the observation of immunoglobulin A (IgA)-positive cells in the conjunctiva was performed by immunohistochemistry. Cytokine concentration in the conjunctiva was determined by the protein-array methods. Messenger RNA expression of T-cell-specific markers, such as T-bet, GATA-3, and Foxp3, in the conjunctiva was detected by reversed transcriptase polymerase chain reaction.The number of eosinophils and CD4-positive lymphocytes increased significantly in the allergy group compared with the control group (P 0.001) but showed no difference between the CTB group and control group. Concentrations of interleukin 4 (IL-4) (P 0.05), B-lymphocyte chemoattractant (P 0.01), and thymus-expressed chemokine (P 0.05) in the conjunctiva were significantly higher in the CTB group than in the other two groups. GATA-3 messenger RNA (mRNA) in the conjunctiva was expressed in the three groups, but T-bet and Foxp3 were not detected.Transconjunctival immunotherapy using CTB can be evaluated by histological examination of eosinophils and CD4-positive T cells, and a mucosal immunity-associated chemokine and a helper T-cell-17-associated chemokine as biomarkers.