Animals ; Antigens, Dermatophagoides ; genetics ; immunology ; Arthropod Proteins ; Asthma ; etiology ; therapy ; Humans ; Mice ; Vaccination ; Vaccines, DNA ; immunology

OBJECTIVETo determine the proportion of Tc17 cells in the lungs of mice with neutrophilic(NEU) asthma, and to investigate the role of Tc17 cells in the pathogenesis of NEU asthma.

METHODSThirty-two C57/B6 mice of clean grade were randomly divided into two groups: NEU asthma and control. The mice in the NEU asthma group were sensitized by airway instillation of ovalbumin (OVA) and lipopolysaccharide (LPS), and challenged with an aerosol of OVA, while those in the control group were sensitized and challenged with normal saline. At 24 hours after the final challenge, bronchoalveolar lavage fluid (BALF) was collected, and the total number and differential counts of nucleated cells and percentage of each type were determined. The lung tissues were separated and hematoxylin-eosin staining was performed to observe the pathological changes of lungs; flow cytometry was applied to determine the percentages of Tc17 and Th17 cells in the lung tissues. Enzyme-linked immunosorbent assay was applied to determine the levels of interleukin-6 (IL-6), transforming growth factor β (TGF-β), and interleukin-17 (IL-17) in BALF.

RESULTSThe NEU asthma group had a significantly higher total number of nucleated cells, a significantly higher percentage of eosinophils, and a significantly higher percentage of neutrophils in BALF than the control group (P<0.01). The NEU asthma group also had significantly higher percentages of Tc17 and Th17 cells than the control group (P<0.01). In the NEU asthma group, the percentage of Tc17 cells was positively correlated with that of Th17 cells (P<0.05). The NEU asthma group had significantly higher concentrations of IL-6, TGF-β, and IL-17 in BALF than the control group (P<0.05).

CONCLUSIONSThe expression of Tc17 cells in the lung tissues increases in mice with NEU asthma, and the increased number of Tc17 cells may be involved in the pathogenesis of NEU asthma. Tc17 cells may play an important role in NEU asthma through IL-17.

Animals ; Asthma ; genetics ; immunology ; CD8-Positive T-Lymphocytes ; immunology ; Female ; Humans ; Interferon-gamma ; immunology ; Interleukin-17 ; genetics ; immunology ; Interleukin-4 ; genetics ; immunology ; Interleukin-6 ; genetics ; immunology ; Lymphocyte Count ; Mice ; Th17 Cells ; immunology

BACKGROUNDRecent studies have shown that T helper type-2 (Th2) cells can induce the apoptosis of CD4+CD25+ Treg cells or resist the immunosuppressive effect of Treg cells. We hypothesize that an imbalance of Th2/Treg is present in patients with allergic asthma.

METHODSTwenty-two patients with mild asthma, 17 patients with moderate to severe asthma, and 20 healthy donors were enrolled. All patients were allergic to house dust mites. The proportion of peripheral blood CD4+CD25+ Treg cells and Th2 cells were determined by flow cytometry. The concentration of interleukin (IL)-10, transforming growth factor (TGF)-β and IL-4 in plasma was determined by enzyme linked immunosorbent assay. In these subjects, peripheral blood mononuclear cells from 17 mild asthmatic patients, 13 moderate to severe asthmatic patients and 14 healthy donors were acquired and expression of forkhead box P3 (Foxp3) and GATA-3 mRNA was detected by reverse-transcriptase polymerase chain reaction.

RESULTSCompared with healthy donors and patients with mild asthma, the percent of CD4+CD25+ Treg cells and plasma IL-10 levels were decreased in patients with moderate to severe asthma. There were no significant differences in Foxp3 mRNA expression among three groups, but a downward trend seen among patients with asthma. However, the percent of Th2 cells, IL-4 levels and expression of GATA-3 mRNA was markedly higher in patients with mild and moderate to severe asthma than in the control group. The ratio of Th2/Treg and their cytokines was increased in allergic asthma, especially for moderate to severe asthma. The ratio of GATA-3/Foxp3 mRNA was also increased in allergic asthma. In patients with moderate to severe asthma, the percentage of peripheral blood Treg cells was negatively correlated to the percentage of Th2 cells and IL-4 levels.

CONCLUSIONSThe decline of CD4+CD25+ Treg cells in patients with moderate to severe asthma may play an important role in progress of the disease. Furthermore, the deficiency of CD4+CD25+ Treg cells was associated with the over-expression of Th2 response.

Asthma ; etiology ; immunology ; Cytokines ; blood ; Forkhead Transcription Factors ; genetics ; GATA3 Transcription Factor ; genetics ; Humans ; RNA, Messenger ; analysis ; T-Lymphocytes, Regulatory ; immunology ; Th2 Cells ; immunology

OBJECTIVETo observed efficacy differences of acupuncture at "Zusanli" (ST 36) in rats with asthma and asthma with spleen-deficency, so as to investigate the therapeutic mechanism.

METHODSSixty SD rats were randomly divided into 5 groups according to their weight, named as an asthma with spleen-deficency group (group A), an acupuncture on asthma with spleen-deficency group (group B), an asthma group (group C), an acupuncture on asthma group (group D) and a control group. The rat models with spleen-deficiency in the first two groups were set up by TCM, then the rats of asthma model in the first four groups were induced by egg albumin, but the control group was treated by the same dose of saline. The group B and the group D were both treated with acupuncture at "Zusanli" (ST 36), once each day for 8 days, and the other groups remained unhandled. The mRNA expressions of Fas and Bcl-2 in the lung tissues were detected by hybridization in situ and apoptosis was detected by TUNEL (terminal dexynucleotidyl transferase-mediated dutp nick end labeling).

RESULTSCompared with the control group, in both the group A and the group C, the expression of Fas mRNA significantly decreased, but the expression of Bcl-2 mRNA significantly increased (all P < 0.01), and eosinophils (EOS) counts significantly increased, but EOS apoptosis rate significantly decreased (all P < 0.01). Compared with the group C, in the group A, the expressions of Fas mRNA significantly decreased, but the expressions of Bcl-2 mRNA and EOS counts significantly increased (all P < 0.01). At the same time, compared with the corresponding asthma groups, in both acupuncture groups, Fas mRNA expression obviously increased, Bcl-2 mRNA expression was significantly reduced (all P < 0.01), EOS counts remarkably decreased and EOS apoptosis rate significantly increased (all P < 0.01). There were no significant differences in the expressions of Fas mRNA and Bcl-2 mRNA between the two acupuncture groups (both P > 0.05), but compared with group B,in the group D, EOS counts significantly decreased and EOS apoptosis rate significantly increased (both P < 0.01).

CONCLUSIONAcupuncture at "Zusanli" (ST 36) can regulate the disorders of Fas mRNA and Bcl-2 mRNA expression in the state of both asthma and asthma with spleen-deficency, promote EOS apoptosis so as to inhibit the development of inflammatory reaction of asthma, showing that acupuncture at "Zusanli" (ST 36) has certain advantages on regulation of related gene of EOS in asthma with spleen-deficency.

Acupuncture Points ; Acupuncture Therapy ; Animals ; Apoptosis ; Asthma ; genetics ; immunology ; physiopathology ; therapy ; Eosinophils ; cytology ; immunology ; Gene Expression ; Humans ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; fas Receptor ; genetics ; immunology

OBJECTIVEInterleukin-4 plays a key role in the development of asthma. Overseas studies have shown that Q576R polymorphism in the interleukin-4 receptor (IL-4R) gene is related to asthma as well as increased serum IgE levels. This study was designed to investigate the association of Q576R polymorphism in IL-4R gene with childhood asthma and serum IgE levels.

METHODSThe polymorphism of IL-4R Q576R was determined by PCR/RFLP and serum total IgE level was measured using ELISA in 94 children with asthma. Sixty-eight healthy children served as controls.

RESULTSThe distribution frequency of heterozygous genotype Q576R (41%) and mutant allele R576 (26%) was significantly higher in children with asthma than that of controls (16% each) (P < 0.01; P < 0.05). The total serum IgE level between patients with genotype Q576R and Q576Q was not significantly different (225.78 +/- 51.43 IU/mL vs 163.24 +/- 31.32 IU/mL, P> 0.05).

CONCLUSIONSThe mutant R576 allele of IL-4R may be one of the candidate genes for susceptibility to asthma. Allele R576 of IL-4R is related to asthma but is irrelevant to the total serum IgE level in children with asthma.

Adolescent ; Asthma ; genetics ; immunology ; Child ; Child, Preschool ; Female ; Humans ; Immunoglobulin E ; blood ; Male ; Polymorphism, Genetic ; Receptors, Interleukin-4 ; genetics

OBJECTIVETo investigate the preventive effect of interleukin-18 (IL-18) gene modified mature dendritic cells (mDC) vaccine on airway inflammation in mouse asthma model.

METHODSThe asthma model was induced by injection of ovalbumin (OVA) in BALB/c mice. IL-18 gene modified mouse mature dendritic cells (mDC) were detected by flow cytometry and its capacity of inducing allogeneic T cell responses was examined by mixed lymphocyte reaction (MLR). The OVA-induced asthmatic mice were randomly divided into 6 groups: PBS group, DXM group, mDC group, Ad-LacZ-mDC group, Ad-IL-18-mDC group and control group. The pathological changes in lung tissues were assayed by HE and AB-PAS staining. The numbers of inflammatory cells and percentage of eosinophils (EOS) in bronchoalveolar lavage fluid (BALF) were counted. The levels of IFN-γ IL-4 and IL-13 in culture supernatant of splenocytes were measured by ELISA method. The percentage of CD4(+)CD25(+)Foxp3(+) Treg was assessed by flow cytometry analysis.

RESULTThe vaccine was effective in decreasing the infiltration of EOS and accumulation of airway goblet cells in lung tissues, the numbers of inflammatory cells and percentage of EOS in BALF, and the levels of IL-4 and IL-13 in culture supernatant of splenocytes, and in increasing the levels of IFN-γ in culture supernatant of splenocytes and the percentage of CD4(+)CD25(+)foxP3(+) reg.

CONCLUSIONIL-18 gene modified mDC vaccine has a preventive effect on airway inflammation in OVA-induced asthmatic mice.

Animals ; Asthma ; immunology ; pathology ; prevention & control ; Dendritic Cells ; immunology ; Disease Models, Animal ; Genetic Therapy ; Interleukin-18 ; genetics ; Lung ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Ovalbumin ; immunology

Asthma ; immunology ; virology ; Bronchiolitis, Viral ; complications ; immunology ; virology ; Child ; Humans ; Respiratory Syncytial Virus Infections ; complications ; genetics ; immunology ; Risk Factors ; Severity of Illness Index

OBJECTIVETo observe the effects of bacterial lysates (OM-85BV) and all trans-retinoic acid (ATRA) on airway inflammation in asthmatic mice, and to investigate the immunoregulatory mechanism of OM-85BV and ATRA for airway inflammation in asthmatic mice.

METHODSForty female BALB/c mice were randomly divided into five groups: normal control, model, OM-85BV, ATRA, and OM-85BV+ATRA. A bronchial asthma model was established by intraperitoneal injection of ovalbumin (OVA) for sensitization and aerosol challenge in all mice except those in the normal control group. On days 25-34, before aerosol challenge, the model, OM-85BV, ATRA, and OM-85BV+ATRA groups were given normal saline, OM-85BV, ATRA, and OM-85BV+ATRA respectively by gavage. Normal saline was used instead for sensitization, challenge, and pretreatment before challenge in the normal control group. These mice were anesthetized and dissected at 24-48 hours after the final challenge. Bronchoalveolar lavage fluid (BALF) was collected from the right lung to measure the levels of interleukin-10 (IL-10) and interleukin-17 (IL-17) by ELISA. The left lung was collected to observe histopathological changes by hematoxylin-eosin staining. The relative expression of ROR-γT mRNA was measured by quantitative real-time PCR.

RESULTSCompared with the normal control group, the model group showed contraction of the bronchial cavity, increased bronchial secretions, and a large number of infiltrating inflammatory cells around the bronchi and alveolar walls, as well as a significantly reduced level of IL-10 (P<0.05) and significantly increased levels of IL-17 and ROR-γT mRNA (P<0.05). Compared with the model group, the OM-85BV, ATRA, and OM-85BV+ATRA groups showed a significant reduction in infiltrating inflammatory cells around the bronchi and alveolar walls; the OM-85BV group showed a significant increase in the level of IL-10 in BALF (P<0.05) and significant reductions in the levels of IL-17 and ROR-γT mRNA (P<0.05); the ATRA group showed significant reductions in the levels of IL-17 and ROR-γT mRNA (P<0.05). Compared with the OM-85BV group, the OM-85BV+ATRA group had significantly increased relative expression of ROR-γT mRNA (P<0.05). Compared with the ATRA group, the OM-85BV+ATRA group had significantly increased levels of IL-10 and IL-17 in BALF (P<0.05).

CONCLUSIONSBoth OM-85BV and ATRA can reduce respiratory inflammation in asthmatic mice. However, a combination of the two drugs does not have a better effect than them used alone.

Animals ; Asthma ; drug therapy ; genetics ; immunology ; Cell Extracts ; administration & dosage ; Female ; Humans ; Interleukin-10 ; genetics ; immunology ; Interleukin-17 ; genetics ; immunology ; Lung ; drug effects ; immunology ; Mice ; Mice, Inbred BALB C ; Tretinoin ; administration & dosage

OBJECTIVETo investigate the effect of triggering receptor expressed on myeloid cells 2 (TREM-2) overexpression on airway inflammation and remodeling in mice with asthma.

METHODSA total of 40 BALB/c mice were randomly divided into normal control, asthma, empty vector, and TREM-2 overexpression groups (n=10 each). Ovalbumin (OVA) sensitization and challenge were performed to establish the model of asthma. The mice in the control group were given normal saline, and those in the empty vector and TREM-2 overexpression groups were transfected with adenovirus vector and TREM-2 adenovirus, respectively. RT-PCR and Western blot were used to measure the expression of TREM-2, MMP-2, MMP-9, ADAM33, and ADAM8. Bronchoalveolar lavage fluid (BALF) was collected to perform cell counting and classification. ELISA was used to measure the total serum level of IgE and the levels of cytokines in BALF.

RESULTSCompared with the control group, the asthma group showed significant reductions in the mRNA and protein expression of TREM-2 (P<0.05), a significantly increased level of Th2 cytokine (P<0.05), and significantly increased numbers of total cells and classified cells. Compared with the asthma group, the TREM-2 overexpression group showed a significantly reduced level of Th2 cytokine (P<0.05), a significantly reduced level of IgE (P<0.05), and significantly reduced numbers of total cells and classified cells (P<0.05), as well as significantly downregulated expression of the inflammatory factors and growth factors MMP-2, MMP-9, TGF-β1, ADAM8, and ADAM33 (P<0.05).

CONCLUSIONSTREM-2 overexpression significantly alleviates airway inflammation and airway remodeling in mice with asthma and may become a potential target for the prevention and treatment of childhood asthma.

Airway Remodeling ; Animals ; Asthma ; etiology ; immunology ; Cytokines ; analysis ; Female ; Membrane Glycoproteins ; genetics ; physiology ; Mice ; Mice, Inbred BALB C ; Ovalbumin ; immunology ; RNA, Messenger ; analysis ; Receptors, Immunologic ; genetics ; physiology

PURPOSE: Our previous study indicated that the presence of wheat-specific IgG1 and IgG4 antibodies was associated with work-related symptoms in workers exposed to wheat flour. We performed this study to investigate the genetic polymorphisms of beta2-adrenergic receptors and wheat-specific antibodies in association with the clinical parameters of baker's asthma. MATERIALS AND METHODS: In total, 379 subjects working in a single industrial bakery were enrolled in this study. The skin prick test was performed with common inhalant allergens and wheat flour extract. The presence of serum- specific IgE, IgG1, and IgG4 antibodies to wheat flour were determined by ELISA. Whole blood samples were obtained for genotype analysis. Subjects were genotyped with regard to five candidate single nucleotide polymorphisms (SNPs) of the beta2-adrenergic receptor gene (ADRB2; -47 T>C, 46 A>G, 79 C>G, 252 G>A, and 523 C>A) using a single-base extension method. RESULTS: No significant associations were observed between the genotype/allele frequencies of any of the SNPs tested and any clinical parameters. The haplotype of ADRB2 (GAA composed of 46 A>G, 252 G>A, and 523 C>A) was significantly associated with work-related symptoms (p<0.05). Moreover, in subjects with the AG or GG genotype at 46 A>G and haplotype [GAA] of ADRB2, the prevalence rates of wheat-specific IgG1 antibodies and lower respiratory symptoms increased significantly with exposure intensity (both p<0.05). CONCLUSION: The findings of the present study suggest that ADRB2 genetic polymorphism may contribute to the development of work-related symptoms in workers exposed to wheat flour, which can lead to baker's asthma.
Adult ; Allergens/*immunology ; Asthma/genetics/*immunology ; Enzyme-Linked Immunosorbent Assay ; Female ; *Flour ; Haplotypes ; Humans ; Immunoglobulin G/immunology ; Inhalation Exposure/analysis ; Male ; Occupational Exposure/analysis ; *Polymorphism, Genetic ; Polymorphism, Single Nucleotide ; Receptors, Adrenergic, beta-2/*genetics ; Skin Tests ; Triticum/*immunology