OBJECTIVE: To study the expression of intedeukin-5 (IL-5) mRNA and protein in Nasal polyps and in the inferior turbinate, and to explore the relationship between the expression and the Clinical features. METHOD: Real time fluorescent quantitative PCR(RT-qPCR) and immunohistochemical staining were used to detect the expression of intedeukin-5 (IL-5) mRNA and protein in nasal polyps of 24 cases and in inferior turbinate of 15 cases. RESULT: The expression of IL-5 mRNA in Nasal polyps was 7.52 times higher than that in the inferior turbinate (P < 0.01). The expression rate of IL-5 protein in Nasal polyps was 79.17%, significantly higher than that in the inferior turbinate (26.67%) (chi2 = 10.52, P < 0.01). The differece of expression of IL-5 mRNA was not associated with the sexual distinction, unilateral or bilateral nasal polyps and primary or recurrent nasal polyps (P > 0.01), but was associated with the single or multiple nasal polyps, nasal polyps with or without allergic rhinitis and/or asthma. The differece of expression of IL-5 protein was not associated with the sexual distinction, unilateral or bilateral nasal polyps, primary or recurrent nasal polyps, nasal polyps with or without allergic rhinitis and/or asthma, but was associated with the single and multiple nasal polyps. CONCLUSION The high expression of IL-5 mRNA and protein is closely related with the formation edema and development of nasal polyps. Some clinical features of Nasal polyps related to the expression level of IL-5 in nasal polyps.
Female ; Humans ; Interleukin-5 ; genetics ; metabolism ; Male ; Nasal Polyps ; metabolism ; RNA, Messenger ; genetics ; Turbinates ; metabolism

In order to investigate whether Arg110Gln polymorphism in the coding region of the IL-13 gene is associated with asthma and total plasma IgE level in Han nationality in Hubei Chinese population, the allele frequency of 4257(g/a) site and Arg110Gln genotype of IL-13 was detected by using restriction fragment length polymorphism in Han nationality in Hubei Chinese population including 43 asthmatic children, 45 asthmatic adults, 31 control children and 46 control adults. Total plasma IgE was measured by Chemiluminescence assay. The results showed that the frequency of allele A at 4257 bp of IL-13 in children and adults was 0.39 and 0.32, respectively. The GlnGln form of Arg110Gln polymorphism of IL-13 gene was associated with susceptibility of asthma and elevated total plasma IgE in children (P=0.030 and 0.0009, respectively), but not with them in adults (P=0.219 and 0.174, respectively). Our results suggest that the Arg110Gln polymorphism of IL-13 gene is associated with susceptibility of asthma and elevated total plasma IgE in Chinese children of Han nationality in Hubei, but not with them in adults.
Adult ; Asthma ; genetics ; China ; ethnology ; Chromosomes, Human, Pair 5 ; genetics ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; genetics ; Humans ; Immunoglobulin E ; blood ; Interleukin-13 ; genetics ; Male ; Polymorphism, Genetic ; genetics

In order to investigate whether Arg110Gln polymorphism in the coding region of the IL-13 gene is associated with asthma and total plasma IgE level in Han nationality in Hubei Chinese population, the allele frequency of 4257(g/a) site and Arg110Gln genotype of IL-13 was detected by using restriction fragment length polymorphism in Han nationality in Hubei Chinese population including 43 asthmatic children, 45 asthmatic adults, 31 control children and 46 control adults. Total plasma IgE was measured by Chemiluminescence assay. The results showed that the frequency of allele A at 4257 bp of IL-13 in children and adults was 0.39 and 0.32, respectively. The GlnGln form of Arg110Gln polymorphism of IL-13 gene was associated with susceptibility of asthma and elevated total plasma IgE in children (P=0.030 and 0.0009, respectively), but not with them in adults (P=0.219 and 0.174, respectively). Our results suggest that the Arg110Gln polymorphism of IL-13 gene is associated with susceptibility of asthma and elevated total plasma IgE in Chinese children of Han nationality in Hubei, but not with them in adults.
Asthma/*genetics ; China/ethnology ; Chromosomes, Human, Pair 5/genetics ; Gene Frequency ; Genetic Predisposition to Disease/*genetics ; Immunoglobulin E/blood ; Interleukin-13/*genetics ; Polymorphism, Genetic/*genetics

BACKGROUNDExpression of murine calcium-activated chloride channel family member 3 (mCLCA3) has been reported to be increased in the airway epithelium of asthmatic mice challenged with ovalbumin (OVA). However, its role in asthmatic airway inflammation under no OVA exposure has not yet been clarified.

METHODSmCLCA3 plasmids were transfected into the airways of normal BALB/c mice. mCLCA3 expression and airway inflammation in mouse lung tissue were evaluated. Cell differentials and cytokines in bronchoalveolar lavage fluid (BALF) were analyzed. The expression of mCLCA3 protein and mucus protein mucin-5 subtype AC (MUC5AC) were analyzed by Western blotting. The mRNA levels of mCLCA3, MUC5AC and interleukin-13 (IL-13) were determined quantitatively.

RESULTSmCLCA3 expression was not detected in the control group while strong immunoreactivity was detected in the OVA and mCLCA3 plasmid groups, and was strictly localized to the airway epithelium. The numbers of inflammatory cells in lung tissue and BALF were increased in both mCLCA3 plasmid and OVA groups. The protein and mRNA levels of mCLCA3 and MUC5AC in the lung tissue were significantly increased in the mCLCA3 plasmid and OVA groups compared to the control group. The level of IL-13, but not IL-4, IL-5, IFN-γ, CCL2, CCL5 or CCL11, was significantly increased compared with control group in BALF in the mCLCA3 plasmid and OVA groups. The level of IL-13 in the BALF in the mCLCA3 plasmid group was much higher than that in the OVA group (P < 0.05). The level of mCLCA3 mRNA in lung tissue was positively correlated with the levels of MUC5AC mRNA in lung tissue, IL-13 mRNA in lung tissue, the number of eosinophils in BALF, and the content of IL-13 protein in BALF. The level of IL-13 mRNA in lung tissue was positively correlated with the number of eosinophils in BALF and the level of MUC5AC mRNA in lung tissue.

CONCLUSIONThese findings suggest that increased expression of a single-gene, mCLCA3, could simulate an asthma attack, and its mechanism may involve mCLCA3 overexpression up-regulating IL-13 expression.

Allergens ; Animals ; Asthma ; Chloride Channels ; Female ; Inflammation ; chemically induced ; metabolism ; Interleukin-13 ; metabolism ; Interleukin-4 ; genetics ; metabolism ; Interleukin-5 ; genetics ; metabolism ; Mice ; Mice, Inbred BALB C ; Mucin 5AC ; genetics ; metabolism ; Ovalbumin ; pharmacology

OBJECTIVETo investigate the correlation between nuclear factor-kappa B (NF-kappaB) activity and cytokine expression in nasal mucosa of chronic sinusitis.

METHODSIL-5, IL-6 and IL-8 levels in nasal mucosa were assayed by the method of ELISA in 52 cases of chronic sinusitis [concomitant with allergic rhinitis (AR group), without allergic rhinitis (NAR group)] and 12 normal subjects. Semi-quantitative RT-PCR and immunohistochemical staining were used to examine P50 and P65 subunits of NF-KB expressions and activation in nasal mucosa. The correlation between activities of NF-KB P50 and P65 subunits and cytokine expression was evaluated.

RESULTSIL-5, IL-6 and IL-8 levels in both AR and NAR groups were significantly increased (all P < 0.01 for AR group; P < 0.05, 0.05, 0.01, respectively, for NAR group, as compared with normal group), and the levels were much higher in AR group than that in NAR group (P < 0.01, 0.05, 0.01, respectively). The levels of P50 and P65 mRNA in both AR and NAR groups were enhanced (all P < 0.01 for AR group; all P < 0.01 for NAR group, as compared with normal group), and AR group had markedly greater P50 and P65 mRNA levels in comparison with NAR group (both P < 0.05). Immunohistochemical study revealed that nucleus-present rates of P50 and P65 in both AR and NAR groups were significantly higher than those of control group (all P < 0.01), and they were much greater in AR group as compared with NAR group (all P < 0.01). Pearson correlation analysis demonstrated that P50 and P65 nucleus-present rates were closely correlated with IL-6 and IL-8 levels, but not IL-5. The correlation coefficient was 0. 49 for P50 and IL-6, 0. 54 for P50 and IL-8, 0. 61 for P65 and IL-6, and 0.66 for P65 and IL-8 (all P < 0.01).

CONCLUSIONSActivation of P50 and P65 subunits of NF-kappaB might be one of the mechanisms for induction of IL-6 and IL-8 expression in chronic sinusitis. Concomitance of allergic rhinitis with chronic sinusitis further increased activities of NF-kappaB subunits, and further elevated IL-6 and IL-8 expression. IL-5 expression was independent of NF-kappaB pathway in chronic sinusitis.

Adult ; Chronic Disease ; Female ; Humans ; Interleukin-5 ; metabolism ; Interleukin-6 ; metabolism ; Interleukin-8 ; metabolism ; Male ; Middle Aged ; NF-kappa B p50 Subunit ; metabolism ; Nasal Mucosa ; metabolism ; RNA, Messenger ; genetics ; Rhinitis ; metabolism ; Sinusitis ; metabolism ; Transcription Factor RelA ; metabolism

Toxocariasis is a soil-transmitted helminthozoonosis due to infection of humans by larvae of Toxocara canis. The disease could produce cognitive and behavioral disturbances especially in children. Meanwhile, in our modern era, the incidence of immunosuppression has been progressively increasing due to increased incidence of malignancy as well as increased use of immunosuppressive agents. The present study aimed at comparing some of the pathological and immunological alterations in the brain of normal and immunosuppressed mice experimentally infected with T. canis. Therefore, 180 Swiss albino mice were divided into 4 groups including normal (control) group, immunocompetent T. canis-infected group, immunosuppressed group (control), and immunosuppressed infected group. Infected mice were subjected to larval counts in the brain, and the brains from all mice were assessed for histopathological changes, astrogliosis, and IL-5 mRNA expression levels in brain tissues. The results showed that under immunosuppression, there were significant increase in brain larval counts, significant enhancement of reactive gliosis, and significant reduction in IL-5 mRNA expression. All these changes were maximal in the chronic stage of infection. In conclusion, the immunopathological alterations in the brains of infected animals were progressive over time, and were exaggerated under the effect of immunosuppression as did the intensity of cerebral infection.
Animals ; Brain/*pathology ; Disease Models, Animal ; Female ; Gene Expression Profiling ; Histocytochemistry ; *Immunocompromised Host ; Immunohistochemistry ; Interleukin-5/genetics ; Male ; Mice ; Parasite Load ; Toxocara canis/*immunology ; Toxocariasis/*immunology/*pathology

Toxocariasis is a soil-transmitted helminthozoonosis due to infection of humans by larvae of Toxocara canis. The disease could produce cognitive and behavioral disturbances especially in children. Meanwhile, in our modern era, the incidence of immunosuppression has been progressively increasing due to increased incidence of malignancy as well as increased use of immunosuppressive agents. The present study aimed at comparing some of the pathological and immunological alterations in the brain of normal and immunosuppressed mice experimentally infected with T. canis. Therefore, 180 Swiss albino mice were divided into 4 groups including normal (control) group, immunocompetent T. canis-infected group, immunosuppressed group (control), and immunosuppressed infected group. Infected mice were subjected to larval counts in the brain, and the brains from all mice were assessed for histopathological changes, astrogliosis, and IL-5 mRNA expression levels in brain tissues. The results showed that under immunosuppression, there were significant increase in brain larval counts, significant enhancement of reactive gliosis, and significant reduction in IL-5 mRNA expression. All these changes were maximal in the chronic stage of infection. In conclusion, the immunopathological alterations in the brains of infected animals were progressive over time, and were exaggerated under the effect of immunosuppression as did the intensity of cerebral infection.
Animals ; Brain/*pathology ; Disease Models, Animal ; Female ; Gene Expression Profiling ; Histocytochemistry ; *Immunocompromised Host ; Immunohistochemistry ; Interleukin-5/genetics ; Male ; Mice ; Parasite Load ; Toxocara canis/*immunology ; Toxocariasis/*immunology/*pathology

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

The acute and chronic respiratory tract inflammation models were made to investigate the effect and mechanism of sterol extracts from Begonia Sinensis Rhizome (BSR). The first model of acute lung injury was made with Kunming mice by inhaling cigarette smoke, then the mice were treated with different concentrations of BSR sterol extracts. Lung tissue morphology was detected by HE staining, TNF-alpha/MPO were detected by Elisa, and cPLA2 protein were, detected by Western blotting respectively. Results showed that in model group, lung sheet became real, alveolar space shrank or disappeared, alveolar septum was thickened, plenty of inflammatory cells were infiltrated, capillary blood vessels were congestive and the expression of TNF-α, MPO, cPLA2 increased; after administration, a small amount of inflammatory cells were infiltrated, alveolar septum became obvious, capillary congestion status was significantly relieved and the expression of TNF-α, MPO, cPLA2 decreased (P < 0.05). The second model of chronic respiratory tract inflammation in BALB/c mice with bronchial asthma was induced by OVA, then the mice were treated with different concentrations of BSR sterol extracts. Lung tissue morphology was detected by HE staining, indexes such as IL-4, IL-5, IL-13 were detected by Elisa, and the cPLA2 protein expression was detected by Western blotting respectively. Results showed that in model group, a lot of inflammatory cells around lung vessels and bronchi exuded, bronchial goblet cells proliferated and the expression of IL-4, IL-5, IL-13, cPLA2 increased; after administration, inflammatory and goblet cell hyperplasia reduced, the expression of IL-4, IL-5, IL-13, cPLA2 also decreased (P < 0.05). The above results showed BSR sterol extracts could resist against respiratory inflammation by inhibiting cPLA2 in a dose-dependent manner.
Animals ; Asthma ; drug therapy ; genetics ; immunology ; Begoniaceae ; chemistry ; Cytokines ; genetics ; immunology ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Interleukin-13 ; genetics ; immunology ; Interleukin-4 ; genetics ; immunology ; Interleukin-5 ; genetics ; immunology ; Lung ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Rhizome ; chemistry ; Sterols ; administration & dosage

BACKGROUNDAsthma is clinically related with the degree of eosinophilic inflammation. How asthmatic airway inflammation is affected is still poorly understood. So the effects of bone marrow-derived hematopoietic cells expressing CD(34) (CD(34)(+)) and interleukin-5 (IL-5) receptor messenger RNA (IL-5R mRNA+) on asthmatic airway inflammation were investigated.

METHODSBalb/c mice were sensitized and challenged by ovalbumin (OVA) to establish an asthmatic model while control mice were sensitized and exposed to sterile saline. The mice were killed at different time points after being challenged by OVA and sterile saline. Then, bronchoalveolar lavage fluid (BALF), peripheral blood (PB) and bone marrow (BM) were prepared. Eosinophils in PB (PBEOS) and BALF (BALFEOS), nuclear cells in BALF, PB and BM were counted. By flow cytometry, the percentage of CD(34)(+) cells to nucleated cells in PB, BM and the relative number of CD(34)(+) cells in PB (PBCD(34)(+)) and BM (BMCD(34)(+)) were calculated. Immunocytochemistry and in situ hybridization were used to investigate the hematopoietic cells with co-localized expression of CD(34) and IL-5R mRNA in BM (BMCD34+IL-5R mRNA+). The percentage of BMCD34+IL-5R mRNA+ to BMCD(34)(+) was calculated.

RESULTSTwelve hours after challenge by OVA, BALFEOS and PBEOS in the experimental group were significantly higher than those in the control group (P < 0.01). Twenty-four hours after OVA challenge, BALFEOS, PBEOS and BMCD34+IL-5R mRNA+ were elevated maximally, significantly different from those in the control group (P < 0.01). Forty-eight hours after OVA challenge, BALFEOS and BMCD34+IL-5R mRNA+ were still significantly higher than those of the controls (P < 0.01). The other markers reverted to normal. In 60 mice, BMCD34+IL-5R mRNA+ was closely correlated with the BALEOS, PBEOS, BMCD(34)(+) and BMCD(34)(+) (%) (P < 0.05).

CONCLUSIONSThe amount of CD(34)(+) cells expressing IL-5R mRNA increased in the BM of asthmatic model mice, which favors eosinophilopoiesis and eosinophilic airway inflammation. A signal pathway exists between the lungs and the bone marrow, which is involved in the initiation and maintenance of asthmatic airway inflammation.

Animals ; Antigens, CD34 ; analysis ; Asthma ; immunology ; Bone Marrow Cells ; cytology ; Bronchoalveolar Lavage Fluid ; cytology ; Inflammation ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; RNA, Messenger ; analysis ; Receptors, Interleukin ; genetics ; Receptors, Interleukin-5