Effect of RNA interference therapy on the mice eosinophils CCR3 gene and granule protein in the murine model of allergic rhinitis

OBJECTIVE@#To observe the clinical manifestations of allergic rhinitis mice and the expression changes of the eosinophils CCR3 and the granule protein mRNA in the bone marrow, peripheral blood and nasal lavage fluid.@*METHODS@#Twenty-four BALB/c mice were randomly divided into the control group, PBS therapy group, siRNA therapy group and the CCR3 siRNA therapy group (n=6). Allergic rhinitis model were sensitized and stimulated by ovalbunfin, and CCR3 siRNA therapy group were administered with CCR3 transnasally before stimulated. The levels of the eosinophils CCR3, MBP, ECP and EPO in bone marrow, peripheral blood and nasal lavage fluid were detected by RT-PCR.@*RESULTS@#Compared to the control group and CCR3 siRNA therapy group, the nasal mucosa of the PBS therapy group and siRNA therapy group developed epithalaxy, goblet cells hyperplasia, squamous epithelium metaplasia, epithelium necrosis, lamina propria and submucosa gland hyperplasia, vasodilatation, tissue edema, and the characterized eosinophil infiltration. RT-PCR indicated that the CCR3 mRNA, MBP, ECP and EPO expression in bone marrow, peripheral blood and nasal lavage fluid of the CCR3 siRNA therapy group was lower than the PBS therapy group and siRNA therapy group (P<0.05).@*CONCLUSIONS@#The RNA interference therapy to CCR3 by local administration pernasal can suppress the process of the development, migration and invasion of the allergic rhinitis eosinophil, thus can reduce the effect of eosinophils and then reduce the inflammation effect of the allergic rhinitis. It may be a new treatment for respiratory tract allergic inflammation.

Apoptosis and eosinophils. Regulation and clinical relevance

Apoptosis, or program cell death, is a process of fundamental biological importance, and eosinophil apoptosis is believed to be the primary mechanism for removing eosinophils from the lung followed by their recognition and phagocytosis by macrophages or resident bronchial epithelial cells. There is, therefore, an increased interest in the fundamental role of the signals and intracellular signaling molecules that initiate and control apoptosis in human eosinophils though much remains to be established. This article reviews briefly the cross talks between apoptosis and eosinophils and summarizes the recent developments in this field

Relationship between Dendritic Cells and Activated Eosinophils in Induced Sputum of Asthmatics

It has been suggested that dendritic cells (DCs) are critical antigen presenting cells for eosinophilic airway inflammation in a mouse model of asthma, and cysteinyl leukotrienes may play a role in DC trafficking in asthmatics. We investigated whether the number of DCs is increased in the induced sputum of both atopic and nonatopic asthmatics and is related to activated eosinophil count in the sputum. Sputum was induced by inhalation of hypertonic saline in 9 atopic and 12 nonatopic asthmatics and 10 nonatopic normal controls, and differential cell counts were performed. DCs and activated eosinophils were identified by immunocytochemistry with monoclonal antibodies (anti-CD1a and EG2, respectively). There were significantly higher percentages of eosinophils, EG2+ cells, and CD1a+ DC in the sputum of atopic and nonatopic asthmatics compared with normal controls, respectively. In asthmatics, the percentage of CD1a+ DC was significantly correlated with that of EG2+ cells (Rs=0.62, p=0.004). We demonstrated that the increased number of DCs was evident in the induced sputum of both atopic and nonatopic asthmatics, and the DC number was related to the activated eosinophil count, which suggests that DCs may contribute to the ongoing eosinophilic inflammation in asthmatic airways, and vice versa.

Oral disodium cromoglycate and ketotifen for a patient with eosinophilic gastroenteritis, food allergy and protein-losing enteropathy.

We present a case report of a 10 years old boy with protein-losing enteropathy and eosinophilic gastroenteritis who had positive histamine release tests, increased allergen-specific IgE antibodies to some food items, and low levels of total serum protein and albumin. Upper gastrointestinal endoscopy revealed a number of polyps and diffuse gastritis. Biopsy specimens of the stomach and duodenum showed widespread eosinophilia and neutrophilia. Although a restricted diet was recommended, a diet which excluded foods with positive results to both histamine release test and allergen-specific IgE antibodies was poorly tolerated, and the patient rejected systemic administration of corticosteroids. Thus, we initiated an oral disodium cromoglycate (DSCG) and ketotifen therapy. After oral DSCG and ketotifen administration, the patient's condition improved gradually. Therefore, oral DSCG and ketotifen therapy might be considered as treatment option in patients with eosinophilic gastroenteritis and protein-losing enteropathy caused by food allergy.

The relationship between IL-13 gene polymorphism and the levels of serum IL-13 and serum eosinophil cation protein in asthmatic children / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To explore the relationship between IL-3 gene polymorphism and the levels of serum IL-3 and eosinophil cation protein (ECP) for understanding the role of IL-3 gene polymorphism in the mechanism of childhood asthma.</p><p><b>METHODS</b>The method of restriction fragment length polymorphism (RFLP) was adopted in detecting +1923 site polymorphism of IL-13 gene in intron 3 region, ELISA was employed in detecting the level of serum IL-13, and fluorescent enzyme-linked immunoassay was used to detect the level of serum ECP.</p><p><b>RESULTS</b>The frequency distribution of TT, TC genotypes of IL-13 Intron 3+1923 site in asthmatic children was higher than that of CC genotype in normal control (P<0.05), and the levels of serum IL-13 and ECP of TT, TC genotypes were significantly higher than those of CC genotype respectively (P<0.01).</p><p><b>CONCLUSION</b>The close relationship of IL-3 gene polymorphism with the levels of serum IL-13 and ECP suggests that IL-3 gene polymorphism may play an important role in the mechanism of childhood asthma.</p>

Effects of interleukin-1 receptor antagonist on the apoptosis of eosinophil in guinea pig with asthma / 药学学报

<p><b>AIM</b>To evaluate the effect of interleukin-1 receptor antagonist(IL-1ra) on apoptosis and associated mechanism of eosinophil in guinea pig with asthma.</p><p><b>METHODS</b>A model of guinea pig with asthma was established. After inhalation of different concentrations of IL-1ra, asthma was induced in the guinea pig for 8 days, the concentration of eosinophil cationic protein (ECP) in serum and bronchoalveolar lavage fluid (BALF), IL-5 in serum, the eosinophil counts and apoptosis were assayed by radioimmunology, enzyme-linked immunosorbent assay(ELISA), fluoromicroscope and light microscope.</p><p><b>RESULTS</b>IL-1ra indirectly decreased the level of IL-5 in serum, improved the apoptosis of eosinophil(EOS) in lung, then decreased the level of ECP in serum and BALF.</p><p><b>CONCLUSION</b>Inhalation of nebulized IL-1ra showed protective effect against asthma through change of the activity and infiltration of EOS in lung.</p>

Effect of pingchuan mixture on eosinophil cation protein and interleukin-5 in experimental guinea pigs with asthma / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the effect of Pingchuan Mixture (PCM) on plasma eosinophil cation protein (ECP), interleukin-5 in bronchial alveolar lavage fluid (BALF) and inflammatory cell count in experimental guinea pigs with asthma.</p><p><b>METHODS</b>The eosinophil, neutrophil, lymphocyte count were conducted by conventional method, IL-5 was detected by ELISA and ECP determined by RIA.</p><p><b>RESULTS</b>Levels of eosinophil, neutrophil, lymphocyte, ECP and IL-5 after treatment were significantly lower than those before treatment, the difference between groups treated respectively by PCM, aminophylline, dexamethasone and Dingchuan Zhike Tablet was insignificant.</p><p><b>CONCLUSION</b>PCM could treat asthma by reducing the inflammatory cell count, ECP and IL-5.</p>

Role of IgG, IgA, and IgE Antibodies in Nasal Polyp Tissue: Their Relationships with Eosinophilic Infiltration and Degranulation

To confirm local production of IgE, and evaluate role of immunoglobulins on eosinophil activation in nasal polyp (NP) tissue, we measured IgG, IgA, secretory IgA(sIgA), total (tIgE) and specific IgE (sIgE) to Dermatophagoides pteronyssinus(DP) by ELISA in NP tissue homogenates from 51 subjects. They were classified according to skin reactivity to DP: group I, 15 highly atopic subjects; group II, 18 weakly atopic subjects; and group III, 18 non-atopic subjects. Eosinophil cationic protein (ECP) level was measured by CAP system. Highest level of DP-sIgE was noted in group I, followed by group II and III (p<0.05). Nine (60%) of group I and 4 (22%) of group II subjects had detectable level of DP-sIgE with no significant differences in IgA, sIgA, and IgG. All of NP tissue had eosinophilic infiltration with no significant difference in activated eosinophil count or ECP level among 3 groups. A significant correlation was noted between EG2+ cell count and tIgE (r=0.55, p<0.05), and DP-sIgE level (r=0.60, p<0.05). A significant correlation was also noted between ECP and IgG (r=0.51, p<0.05) and DP-sIgE level (r=0.47, p<0.05) with no significant correlation with IgA or sIgA. These results suggest that DP-sIgE was detectable in NP tissue from weakly atopic subjects as well as highly atopic subjects. IgG and sIgE may have potential roles in eosinophil degranulation in NP tissue.

Correlation of total and specific serum immunoglobulin E levels with the severity of chronic sinusitis in children.

Chronic sinusitis is frequently associated with allergy and asthma. Previous studies have shown that serum immunoglobulin E (IgE) levels correlate with allergy and asthma in adults. Because the role of allergic inflammation in the severity of chronic sinusitis remains controversial in children, we set out to determine whether a correlation exists between serum levels of total and specific IgE and the severity of chronic sinusitis in children. Forty-four children with chronic sinusitis were enrolled in the study. Computed tomographic scans were reviewed and scored for the severity of sinusitis. All children were mite-sensitive. Serum samples were assayed for total IgE and specific IgE antibodies to mite allergen using a fluoroimmunoassay. Fourteen subjects had extensive sinus disease. There was no significant difference in the average of total and specific IgE between the subjects with extensive and limited disease (p = 0.562 and 0.755, respectively). Thirty-four subjects were diagnosed with asthma. The subjects with extensive sinus disease had a higher prevalence of moderate to severe asthma than the subjects with limited disease (p = 0.006), but there was no significant difference in the average of total and specific IgE between the subjects with different severities of asthma. (p = 0.833 and 0.425, respectively). The data suggests that levels of total or specific IgE do not correlate with severity of chronic sinusitis in children. Nonetheless, the severity of chronic sinusitis and asthma correlate well with each other irrespective of total and specific IgE.

Serum eosinophil cationic protein determination in asthmatic children-effect of different collecting tubes used for blood sampling.

We compared the effect of using 2 different serum collecting tubes, serum separation tubes (SST, with clot activator and gel barrier) and conventional glass tubes (with no additives), on circulating levels of eosinophil cationic protein (ECP) in asthmatic children and controls. The serum ECP values obtained from both SST and glass tubes were significantly higher in asthmatic children than in corresponding controls. ECP values were higher in serum samples using SST than in those using glass tubes (P<0.01), while no difference was found between the two in controls. ECP levels correlated with peripheral eosinophil counts, for SST samples and glass tube samples alike. The difference in ECP levels between these two tubes also correlated with circulating eosinophil counts (r = 0.62, P = 0.004) After 18-hour storage at room temperature, the ECP values increased significantly in samples obtained from asthmatic children. No difference in ECP values between SST samples and glass tube samples was found for 18 hour samples. Thus, ECP levels obtained from SST samples and glass tube samples, though reliable, should not be directly compared, especially in asthmatic children with eosinophilia.

Serum Eosinophil Cationic Protein Levels in Patients with Allergic Diseases / 대한임상병리학회지

BACKGROUND: Eosinophil cationic protein (ECP), one of the eosinophil granule proteins released during allergic reactions, may play a major role in the allergic inflammatory process. The measurement of ECP in serum may be a useful indicator of eosinophil activity in ongoing inflammatory processes. We investigated the clinical utility of ECP measurement in serum in patients with bronchial asthma, allergic rhinitis and atopic dermatitis, after standardizing sample processing. METHODS: We measured the serum ECP levels in patients with bronchial asthma (n=38), chronic obstructive pulmonary diseases (COPD) (n=13), respiratory symptoms (n=19), allergic rhinitis (n=26), non-allergic rhinitis (n=24), and atopic dermatitis (n=10) and in normal healthy controls (n=16) by the fluoroenzyme immunoassay using Pharmacia CAP System, and evaluated the correlation between ECP level and blood eosinophil number, or ECP and IgE levels. Blood eosinophil number was counted by the automated cell counter. RESULTS: Serum ECP levels were significantly higher in patients with bronchial asthma (15.6+/- 12.6 g/L), COPD (13.3+/-7.2 g/L), allergic rhinitis (23.8+/-13.2 g/L), and atopic dermatitis (20.6+/- 18.4 g/L) than in normal controls (7.5+/-4.2 g/L) (P <0.05). ECP levels were also significantly higher in patients with bronchial asthma and COPD than in patients with simple respiratory symptoms (6.9+/-4.7 g/L), whose ECP levels did not statistically differ from those in normal controls. ECP levels were also significantly higher in patients with allergic rhinitis than in patients with non-allergic rhinitis (9.5+/-5.1 g/L), whose ECP levels did not statistically differ from those in normal controls. Serum ECP level and eosinophil number in peripheral blood were correlated only in patients with bronchial asthma (r=0.53, P <0.01) and no correlation between ECP and IgE levels was found in all of the patients. CONCLUSIONS: ECP is the one of the secretory components released from the eosinophil granule and measurement of ECP in serum might be one of the noninvasive tool to assess the activity in relation to eosinophil involvement in various allergic diseases.

Toluene Diisocyanate-Induced Allergic Rhinitis in Guinea Pigs: Investigating the Relation between Eosinophil Infiltration and Epithelial Injury

It is well known that many eosinophils are infiltrated in the bronchial and nasal mucosa of allergic patients, and that eosinophil granule proteins can injure the bronchial epithelium. But it is uncertain whether epithelial injury occurs in the nasal mucosa of patients with allergic rhinitis and, if so, whether the injury is related with the eosinophil infiltration. The present study was made with the aim of determining the correlation between eosinophil infiltration and epithelial injury in the nasal mucosa of guinea pigs with experimentally induced nasal allergy. 2,4-Toluene diisocyanate (TDI) is considered to be a causative agent of allergic pulmonary disorder and allergic rhinitis. Guinea pigs were sensitized by applying TDI onto their bilateral nasal vestibules once a day for five consecutive days. Symptom scores, peripheral blood and histopathology of the nasal mucosa in the inferior turbinate were examined in both allergy and control group. The symptom scores were significantly higher in allergy group than in control group and the eosinophils of peripheral blood were found significantly higher in the allergy groups, especially in groups sacrificed 24 and 48 hour after provocation. As well, there was a positive correlation between how heavily infiltrated the eosinophils were and the level of epithelial loss in the nasal mucosa. The correlation was prominent in groups sacrificed 24 and 48 hour after provocation. The findings indicate that the epithelial injury occurs in the nasal mucosa of guinea pigs with nasal allergy and that injury is related to the eosinophil infiltration.