The relationship of interleukin-4 (IL-4) C-33T and C-590T (C-589T) gene polymorphisms with allergic rhinitis was analyzed. Data about the case control studies of IL-4 gene promoter polymorphisms [C-33T and C-590T (C-589T)] and their association with allergic diseases and correlation between serum IL-4 levels and allergic rhinitis were retrieved. The Stata 12.0 statistical software was applied to analyze the correlation between IL-4 gene polymorphisms and allergic rhinitis. The meta-analysis result of TT/CC genotype of -590 (-589) polymorphism showed a significant association with allergic diseases [OR=1.93, 95% CI (1.61-2.31), P=0.00]. Meta-analysis of the TT+TC versus CC genotype of IL-4 C-33/T polymorphism revealed significant associations with allergic diseases [OR=3.23, 95% CI (1.13-9.25), P=0.03]. Meanwhile, there was a significant correlation between serum IL-4 levels and allergic rhinitis [OR=2.52, 95% CI=(1.80-3.23), P=0.00]. IL-4 gene -590 TT genotype may increase the risk of allergic rhinitis and the T allele mutation of -33 might be correlated with allergic rhinitis.
Gene Frequency ; Genotype ; Humans ; Interleukin-4 ; blood ; genetics ; Polymorphism, Single Nucleotide ; Promoter Regions, Genetic ; genetics ; Rhinitis, Allergic ; Rhinitis, Allergic, Perennial ; blood ; genetics ; Risk Factors

IL-28RA is one of the important candidate genes for complex trait of genetic diseases, but there is no published information of the genetic variation in this gene. We scanned the seven exons and their boundary introns sequence of IL-28RA including the promoter regions to analyze genetic variation sites, and identified eighteen single nucleotide polymorphisms (SNPs) and two variation sites. We chose seven SNPs (g.-1193 A>C, g.-30 C>T, g.17654 C>T, g.27798 A>G, g.31265 C>T, g.31911 C>T and g.32349 G>A) of them for large sample size genotyping, and assessed the association of genotype and allele frequencies of these SNPs between allergic rhinitis patients and non-allergic rhinitis controls. We also compared the genotype frequencies between Korean controls and Han Chinese control or Korean Chinese control. We investigated the frequencies of haplotype constructed by these SNPs between allergic rhinitis patients and non-allergic rhinitis controls. Our results suggested that the g.32349 G>A polymorphism of IL-28RA might be associated with susceptibility to allergic rhinitis (P=0.032), but seems to have no relationship with serum total IgE levels. The haplotype frequencies by these SNPs also show significant association between controls and allergic rhinitis patients.
Variation (Genetics) ; Rhinitis, Allergic, Seasonal/blood/*genetics ; Rhinitis, Allergic, Perennial/blood/*genetics ; Receptors, Cytokine/*genetics ; Promoter Regions (Genetics)/genetics ; Polymorphism, Single Nucleotide/*genetics ; Male ; Immunoglobulin E/blood ; Humans ; Haplotypes ; Genotype ; Genetic Predisposition to Disease/genetics ; Gene Frequency ; Female ; Exons/genetics ; Case-Control Studies ; Alleles ; Adult

OBJECTIVETo investigate the distribution characteristics of the single nucleotide polymorphisms (SNPs) of the human CD14 gene in Chinese Han ethnic group children in Wenzhou, and their association with atopic diseases.

METHODSTotally 113 cases were recruited in atopic disease group who met the following criteria: 2 - 12 years old, clinically diagnosed as asthma or allergic rhinitis or atopic dermatitis, elevation of serum total IgE levels and serum specific IgE. Sixty-seven healthy children were enrolled in control group. The related regions of CD14 gene were sequenced to identify and characterize the SNPs, and plasma TIgE and SIgE were detected by immunoassay system and uniCAP system, respectively. The frequency of genotypes and alleles between two groups, as well as the levels of IgE in different genotypes, were compared.

RESULTSCD14/-159 SNP was present in Han ethnic group population of Wenzhou. The frequency of each genotype was 57.0% (TT), 28.0% (TC), 15.0% (CC) in normal children, and 46.9% (TT), 35.4% (TC), 17.7% (CC) in atopic children. No significant difference was found in the distribution of CD14/-159 polymorphism between atopic children and healthy control (chi(2) = 1.918, P > 0.05) according to Hardy-Weinberg principle statistics. There were no significant difference in frequency of each genotype between boys and girls. No significant difference was found in the total plasma IgE levels among groups of TT genotypes [(2520 +/- 460) IU/L], TC genotypes [(2400 +/- 460) IU/L] and CC genotype [(2500 +/- 460) IU/L] (F = 0.807, P > 0.05).

CONCLUSIONCD14/-159 SNP is present in Han ethnic group children in Wenzhou, and other SNP in CD14 gene was not found. TT genotype was the primary genotype in CD14/-159 SNP in the children studied. No relationship between CD14/-159 SNP and atopic disease or serum total IgE level was found.

Asian Continental Ancestry Group ; genetics ; Asthma ; genetics ; Case-Control Studies ; Child ; Child, Preschool ; Dermatitis, Atopic ; genetics ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Immunoglobulin E ; blood ; Lipopolysaccharide Receptors ; genetics ; Male ; Polymorphism, Single Nucleotide ; Rhinitis, Allergic, Perennial ; genetics

OBJECTIVETo investigate whether the single nucleotide polymorphisms (SNP) in the FCER1A gene are associated with serum total IgE level in Chinese allergic rhinitis (AR) cohort.

METHODSA total of 378 patients diagnosed as AR was included in this study. Serum total IgE level was detected by UniCAP100 testing system. A total of 8 representativeness marker SNP which were in FCER1A gene region were selected according to the Beijing people database from Hapmap website and the running results of Haploview 4.1 software. The individual genotyping was performed by MassARRAY platform.

RESULTSThree hundred and seventy-eight individuals (227 males and 151 females) with AR were prospectively recruited. The median of the total serum IgE measurements for the study population were 150 kU/L and ranged from 6.94 kU/L to 5000 kU/L. Levene tests showed the homogeneity of variance of total IgE level among different genotypes regarding each SNP locus. ANOVA tests demonstrated that none of the selected polymorphism loci in FCER1A was associated with total IgE level.

CONCLUSIONPresent study failed to find an association between SNP in the FCER1A gene region and serum total IgE level in Chinese AR.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; Female ; Humans ; Immunoglobulin E ; blood ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Receptors, IgE ; genetics ; Rhinitis, Allergic, Perennial ; blood ; genetics ; Young Adult

OBJECTIVETo study the preparation of recombinant house dust mite group 1 allergen vaccine (chitosan-pVAX1-Derp1 nanoparticles, pVAX1-Derp1/CS) and to investigate the efficacy and mechanism of intranasally given chitosan-pVAX1-Derp1 nanoparticles on mouse model with allergic rhinitis (AR).

METHODSThe chitosan-pVAX1-Derp1 nanoparticles was prepared by complex coacervation, and its nature was identified and analysed. A total of 40 BALB/c rats were randomly divided into 5 groups: the normal group (group A), the AR model group (group B), the chitosan (CS) prevention group (group C), the pVAX1-Derp1 prevention group (group D), and the pVAX1-Derp1/CS prevention group (group E). The nasal cavity of rats in the group B, C, D and E were dripped with phosphate buffered saline (20 µl), CS (20 µl), pVAX1-Der p1 (20 µl), pVAX1-Derp1/CS nanoparticles (20 µl) on the first day and day 8, once daily. Rats in the latter 4 group were sensitized with Der p1 and Al(OH)3 in day 15 and day 22, and challenged with Der p1 to establish AR model from day 36 to day 43, while rats in group A were treated with PBS. Then the level of cytokines in serum was assayed by ELISA, inflammatory reactions in nasal mucosa were analyzed by haematoxylin and eosin staining.

RESULTSpVAX1-Derp1/CS nanoparticles was successfully constructed, the mean grain size of pVAX1-Derp1/CS was (205.3 ± 12.8) nm, and the zeta potential was (30.5 ± 5.6) mV. In nasal mucosa tissue, group B and C showed significant allergic inflammation, while group D and E showed lighter allergic inflammation. Compared with the group B, the group D and E could effectively reduced serum IgE level and IL-4 level [group B: (120.0 ± 8.8) ng/ml, (248.7 ± 10.6) pg/ml; group D: (109.6 ± 14.5) ng/ml, (192.5 ± 10.2) pg/ml; group E: (88.1 ± 8.3) ng/ml, (165.7 ± 9.7) pg/ml; IgE: t value were 3.5, 6.9, all P < 0.01; IL-4: t value were 10.0, 15.2, all P < 0.01], and increased IFN-γ level [group B: (709.0 ± 26.5) pg/ml; group D: (856.3 ± 37.4) pg/ml; group E: (904.8 ± 37.7) pg/ml; t value were 8.2, 10.8, all P < 0.01)]. IL-10 level of group D [(129.9 ± 16.1) pg/ml] and E [(107.1 ± 11.8) pg/ml] was lower than IL-10 level of group B [(160.6 ± 24.2) pg/ml]. The difference were significantly (t value were 2.9, 5.5, all P < 0.05).

CONCLUSIONSChitosan can effectively encapsulate pVAX1-Derp 1 and inhibit nuclease degradation of the plasmid, the DNA vaccine has some preventive effect on AR animal model by nasal immunization.

Administration, Intranasal ; Animals ; Antigens, Dermatophagoides ; immunology ; Chitosan ; administration & dosage ; Cytokines ; blood ; Immunization ; methods ; Immunoglobulin E ; blood ; Mice ; Mice, Inbred BALB C ; Nanoparticles ; administration & dosage ; Nasal Mucosa ; immunology ; Plasmids ; Rhinitis, Allergic, Perennial ; prevention & control ; Vaccines, DNA ; genetics ; immunology

Orai1 is the key subunit of the Ca2+-release-activated Ca2+ channel. Our previous report has demonstrated that Orai1 expression in the airway was upregulated in the ovalbumin (OVA)-induced allergic rhinitis (AR) mouse models. To observe whether inhibition of Orai1 expression in the airway could suppress symptoms in a murine model of AR and to assess the impacts of this inhibition on the responses of local and systemic immunocytes, we administered recombinant lentivirus vectors that encoded shRNA against ORAI1 (lenti-ORAI1) into the nostrils of OVA-sensitized mice before the challenges, and analyzed its effect on allergic responses, as compared with the unsensitized mice and untreated AR mice. Administration of lenti-ORAI1 into the nasal cavity successfully infected cells in the epithelial layer of the nasal mucosa, and significantly decreased the frequencies of sneezing and nasal rubbing of the mice. Protein levels of leukotriene C4, OVA-specific IgE, and IL-4 in the nasal lavage fluid and serum and eosinophil cation protein in the serum were also significantly reduced by lenti-ORAI1, as were the mRNA levels of these factors in the nasal mucosa and spleen. These data suggested that administration of lenti-ORAI1 into the nasal cavity effectively decreased Orai1 expression in the nasal mucosa, alleviated AR symptoms, and partially inhibited the hyperresponsiveness of the local and systemic immune cells including T cells, B cells, mast cells and eosinophils that are involved in the pathogenesis of AR.
Animals ; Calcium Channels/analysis/*genetics/immunology ; *Down-Regulation ; Eosinophil Cationic Protein/blood/genetics ; Glutathione Transferase/blood/genetics/immunology ; Immunoglobulin E/blood/genetics/immunology ; Interleukin-4/blood/genetics/immunology ; Lentivirus/genetics ; Mice ; Mice, Inbred BALB C ; Nasal Mucosa/immunology/metabolism ; Ovalbumin/immunology ; RNA, Messenger/genetics ; RNA, Small Interfering/*administration & dosage/genetics ; Rhinitis, Allergic, Perennial/*genetics/immunology ; Spleen/immunology/metabolism ; *Transfection

BACKGROUNDCurrently anti-inflammatory therapy with steroids for allergic rhinitis need long-term repeated administration, although it is effective. Gene therapy is being suggested to substitute it. The aim of this study was to investigate nonviral vector mediated exogenous gene expression in COS-7 cells in vitro and the effect of intranasal mouse interleukin (mIL)-12 transgene expression on allergen induced eosinophil infiltration of nasal mucosa in a murine model of allergic rhinitis.

METHODSIn vitro COS-7 cells were infected with Epstein-Barr virus (EBV)/lipoplex. The expression of IL-12 p70 in cell culture supernatant was examined by enzyme-linked immunosorbent assay (ELISA). In mice with ovalbumin (OVA) induced allergic rhinitis, EBV/lipoplex was administered by nasal drops before OVA challenge once a day from day 1 to day 10. The expression of IL-12 mRNA and protein, the change of eosinophil count in nasal mucosa and serum total IgE were measured 24 hours after the last challenge.

RESULTSEBV/lipoplex could effectively transfect COS-7 cells. The expression of IL-12 p70 in cell culture supernatant was significantly more than in blank control. IL-12 via EBV plasmid vector transduction could be overexpressed in vivo. In pGEG.mIL-12 treated models, the nasal mucosa revealed a high level of widespread mIL-12 transduction by immunohistochemistry and in situ hybridization. Histological evaluation revealed marked suppression of eosinophil infiltration in nasal mucosa. The eosinophil count in allergic rhinitis group [(26.5 +/- 9.8)/high-power field (HPF)] was significantly increased over control group [(0.40 +/- 0.52)/HPF] (F = 56.94, P < 0.01), while the count in IL-12 gene therapy group [(4.60 +/- 2.63)/HPF] was significantly less than that of allergic group (F = 56.9, P < 0.01). Serum total IgE between in gene therapy mice [(88.83 +/- 6.71) ng/ml] and allergic rhinitis mice [(103.1 +/- 5.7) ng/ml] showed a significant difference (F = 1216, P < 0.05).

CONCLUSIONSNonviral EBV plasmid vector, pGEG.mIL-12 was able to overexpress exogenous gene both in vitro and in murine nasal mucosa in vivo. IL-12 overexpression via EBV/lipoplex could stem allergen induced eosinophil infiltration in nasal mucosa in murine models of allergic rhinitis, which may suggest a new cytokine immunogenetic therapy for allergic rhinitis.

Administration, Intranasal ; Animals ; COS Cells ; Cercopithecus aethiops ; Eosinophilia ; therapy ; Genetic Therapy ; Herpesvirus 4, Human ; genetics ; Immunoglobulin E ; blood ; Interleukin-12 ; genetics ; Lipids ; administration & dosage ; Male ; Mice ; Mice, Inbred BALB C ; Nasal Mucosa ; metabolism ; Rhinitis, Allergic, Perennial ; therapy ; Rhinitis, Allergic, Seasonal ; therapy