OBJECTIVETo evaluate the serological markers and biological marker in the diagnosis of hepatitis E infection in a rhesus monkey model.

METHODS86 rhesus monkeys had been infected with different doses of HEV. Hence, they were taken sequential blood samples at intervals up to 86 weeks for 4 hepatitis E virus (HEV) specific antibody assays (E2-IgM, E2-IgG, GL-IgG, and YES-IgG), and nucleic acid assay.

RESULTSAll the animals produced E2-IgG and all but one also produced E2-IgM and excreted the virus in stool, whereas positive rate of GL-IgG and YES IgG were low and correlated with virus level. Hepatitis occurred over a period of 4 weeks (between 3 an 7 weeks) after infection. Virological marker occurred mainly during incubation period and declined rapidly after onset of hepatitis. Seroconversion of E2-IgM occurred before onset of hepatitis in 70% monkeys and declined rapidly up to 50% of peak value after 4 weeks. E2-IgM seroconversion was closely paralleled by E2-IgG; however, E2-IgG persisted in all animals for the entire duration of experiment of up to 86 weeks. Production of GL-IgG and YES-IgG was delayed by one week after the E2 antibodies, these antibodies showed a transient occurrence and seroprevalence declined to 50% of the peak value over a period of 12 weeks.

CONCLUSIONE2-IgM might be used as a suitable acute hepatitis E marker, and E2-IgG as a suitable epidemiological marker. The seroconversion or titer elevation of GL-IgG and YES-IgG antibodies probably used to confirm the infection. The viral markers are optional for early diagnosis.

Alanine Transaminase ; blood ; Animals ; Biomarkers ; Genotype ; Hepatitis Antibodies ; blood ; Hepatitis E ; diagnosis ; Hepatitis E virus ; classification ; genetics ; immunology ; Immunoglobulin E ; blood ; Immunoglobulin M ; blood ; Macaca mulatta

OBJECTIVETo evaluate anti-HEV diagnostic kits by experimental infecting rhesus monkeys with HEV.

METHODSEight rhesus monkeys were infected with genotype 1 and 4 HEV separately. The alanine aminotransferase (ALT) level of all monkeys were detected before and after the process of infection. HEV RNA in stool specimens was tested by reverse transcriptase-polymerase chain reaction (RT-PCR) assay. Anti-HEV IgG in serum was detected by GL-IgG and WT-IgG.

RESULTSHEV RNA presented in the stool of all the 8 monkeys after infection. The ALT level of 1 monkey infected with genotype 1 HEV and 2 monkeys infected with genotype 4 HEV appeared abnormally after infection. Tested by GL-IgG, 2 of the 4 monkeys infected with genotype 1 HEV and 1 of 4 monkeys infected with genotype 4 HEV seroconverted to anti-HEV IgG. However, when tested by WT-IgG, all the infected monkeys seroconverted to anti-HEV IgG. The anti-HEV IgG tested by WT-IgG was positive during the whole observation period,and the anti-HEV IgG measured by GL-IgG only remained 12 weeks after infection. Detected by GL-IgG and WT-IgG, seropositive conversion of the anti-HEV IgG happened almost at the same time.

CONCLUSIONBoth GL-IgG and WT-IgG could detect the anti-HEV IgG of experimentally infected rhesus monkeys but the WT-IgG had a higher sensitivity for detection of anti-HEV IgG than

Alanine Transaminase ; blood ; Animals ; Disease Models, Animal ; Genotype ; Hepatitis E ; immunology ; virology ; Hepatitis E virus ; genetics ; immunology ; Immunoglobulin G ; immunology ; Macaca mulatta ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVEA deficient interferon-gamma (IFN-gamma) response has been involved in the pathogenesis of severe respiratory syncytial virus (RSV) infection. Gene polymorphisms in IFN-gamma/A+874T have been associated with the susceptibility to asthma and might be related to disease severity of RSV infection. This study investigated the single nucleotide polymorphisms (SNPs) of IFN-gamma/A+874T in Han children in Wenzhou area and to explore the correlation between gene polymorphisms of IFN-gamma/A+874T and the susceptibility and disease severity of RSV bronchiolitis, as well as the effect of SNPs upon nasopharyngeal secretions (NPS) IFN-gamma and total serum IgE levels.

METHODSOne hundred and fourteen hospitalized children with RSV bronchiolitis and 90 healthy controls were recruited. Sequence analysis was used for detecting the SNPs of IFN-gamma/A+874T. NPS IFN-gamma levels were measured using ELISA. Total serum IgE levels were assayed using the chemiluminescence method.

RESULTSIFN-gamma/A+874T gene polymorphisms were present in both the patient and the control groups. AA and AT genotypes were found in both groups, with a AA frequency of 82.5% vs 77.8% and a AT frequency of 17.5% vs 21.1% (p>0.05). The frequency of allele was 90.4% (A) and 9.6% (T) in the patient group, and 88.3% (A) and 11.7% (T) in the control group, respectively. There were no significant differences in the allele frequency between the two groups. Moreover, no difference was found both in NPS IFN-gamma and total serum IgE levels between AA and AT genotypes in the patient group. There were no significant differences in the variation of IFN-gamma/+874 between mild and moderate to severe cases.

CONCLUSIONSIFN-gamma/A+874T gene polymorphisms were present in Han children in Wenzhou area. Gene variations were not associated with the susceptibility and disease severity of RSV bronchiolitis as well as IFN-gamma and total serum IgE levels.

Bronchiolitis ; genetics ; immunology ; Female ; Genetic Predisposition to Disease ; Humans ; Immunoglobulin E ; blood ; Infant ; Interferon-gamma ; genetics ; Male ; Nasopharynx ; immunology ; Polymorphism, Single Nucleotide ; Respiratory Syncytial Virus Infections ; 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

BACKGROUNDAllergen micro-arrays are powerful tools for screening of serum IgE-reactivity. In this study allergen micro-arrays were used to identify dominating IgE-binding allergens and cross-reactivity patterns among selected Chinese allergy patients.

METHODSThe study was conducted using patient sera from the cities of Guangzhou, Nanjing, Chengdu and Shenyang. In total 100 sera with Dermatophagoides pteronyssinus (Der p) specific IgE-levels higher than 50 kU/L were selected for testing against 103 individual allergens.

RESULTSAmong 100 selected patients, 95% showed IgE-reactivity towards house-dust mite allergens Dermatophagoides farinae (Der f) 1, Der f 2 and Der p 2 and 94% were IgE positive against Der p 1, and 60% of sera contained IgE reacting against allergen Euroglyphus maynei (Eur m) 2. IgE against cat allergen, Felisdomesticus (Fel d) 1, was seen in 20%. Only 2% showed specific IgE-reactivity to Der p 10, a panallergen belonging to the tropomyosin family. Serum IgE-reactivity towards other allergens was in general low. IgE-reactivity against pollen allergens showed geographic differences.

CONCLUSIONSThis study clearly confirms that group 1 and group 2 are major allergens of house dust mites. These selected house-dust mite allergy patients are close to being mono-sensitized. Der p 10 is not an important allergen for cross-reactivity. Specific IgE-sensitization towards pollen allergens is low in southern China compared to other regions. The prevalence of food and stinging insect allergens known to give rise to IgE-mediated cross-reactivity is 2% or less.

Adolescent ; Adult ; Allergens ; immunology ; Asian Continental Ancestry Group ; Child ; Child, Preschool ; Female ; Humans ; Hypersensitivity ; blood ; immunology ; Immunoglobulin E ; blood ; genetics ; immunology ; Male ; Middle Aged ; Young Adult

OBJECTIVETo investigate the anti hepatitis E virus (HEV) and HEV RNA in acute and convalescent sera of patients with NonA-E acute hepatitis.

METHODSThe serum samples were taken from 95 patients who were diagnosed as acute NonA-E hepatitis. Enzyme immunoassay (EIA) was used for detecting anti-HEV Immunoglobulin G (IgG, Genolable and Wantai EIA anti-HEV kits). RT-PCR amplification of HEV RNA was based on the open reading frame 2 region of HEV and the PCR products were sequenced.

RESULTSSera from 95 patients who were negative for anti-HEV in acute phase were followed up for 11-35 days to detect the anti-HEV antibody in recovery phase, 16/95 (16.84%) were positive for anti-HEV (wantai EIA anti-HEV kits). Ten (62.50%) were positive for HEV RNA in acute phase. Sequence analysis showed that 4 were HEV genotype. 6 were HEV genotype; 12/95 (12.50%) were positive for anti-HEV (Genolable EIA anti-HEV kits). Seven were positive for HEV RNA; 4 belonged to HEV genotype, 3 were HEV genotype.

CONCLUSIONIt is significant and necessary to detect anti HEV antibody and HEV RNA in patients with HEV infection during acute phase and convalesent phase.

Acute Disease ; Amino Acid Sequence ; Base Sequence ; Convalescence ; Genotype ; Hepatitis Antibodies ; blood ; Hepatitis E ; genetics ; immunology ; virology ; Hepatitis E virus ; genetics ; immunology ; Humans ; Immunoglobulin G ; blood ; RNA, Viral ; blood ; genetics ; Sequence Homology ; Viral Proteins ; genetics

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

OBJECTIVETo investigate the association of the polymorphism of I181L, V183L and E237G in the high affinity immunoglobulin E receptor beta-chain (FcepsilonR1beta) with the susceptibility of childhood asthma and the serum total immunoglobulin E (IgE) level.

METHODSThe coding variants of I181L, V183L and E237G and the serum total IgE level were detected using amplification refractory mutation systemdouble ended arrowpolymerase chain reaction (ARMS-PCR) and double antibody sandwich ELISA respectively in 50 asthmatic children and 40 normal controls from Sichuan Province. The association of gene mutation with the susceptibility of asthma and the serum total IgE level was analyzed.

RESULTSThere were 5 cases of I181L mutation, 2 of V183L mutation, and 7 of E237G mutation in the Asthmatic group. There was no mutation in the Normal control group. The frequency of I181L and E237G mutation in the Asthmatic group were statistically higher than in the Normal control group (P < 0.01). The serum total IgE level in the Asthmatic subgroup with I181L mutation (2.837 +/- 0.407) or E237G mutation (3.044 +/- 0.419) was significantly higher than in the Asthmatic subgroup without gene mutation (2.156 +/- 0.638) and the Normal control group (1.348 +/- 1.291) (P < 0.05 or 0.01).

CONCLUSIONSThe polymorphism of Fc epsilonR1betaI181L and E237G is a susceptible gene of childhood asthma and closely associates with the increased serum total IgE level.

Adolescent ; Asthma ; genetics ; immunology ; Child ; Child, Preschool ; Female ; Genetic Predisposition to Disease ; Humans ; Immunoglobulin E ; blood ; Infant ; Male ; Mutation ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Receptors, IgE ; genetics

OBJECTIVETo study the roles of CD4(+)CD25(+) regulatory T cells and Foxp3 mRNA in peripheral blood as well as serum total immunoglobulin E (IgE) in the pathogenesis of bronchiolitis caused by respiratory syncytial virus (RSV).

METHODSThe proportion of CD4(+)CD25(+) regulatory T cells and expression of Foxp3 mRNA in peripheral blood, and total serum IgE level were tested by flow cytometry, RT-PCR and ELISA respectively in 57 children with RSV bronchiolitis (26 atopic patients and 31 nonatopic patients). Twenty five healthy children were used as the control group.

RESULTSThe proportion of CD4(+)CD25(+) regulatory T cells in peripheral blood in children with bronchiolitis, either in the atopic (7.7+/- 1.6%)or the nonatopic group (8.8+/- 2.1%), was significantly lower than that in the control group (10.5+/- 1.6%) (P< 0.01). Foxp3 mRNA expression in peripheral blood was significantly lower in both atopic and nonatopic children with bronchiolitis than that in the control group (P< 0.01). Significantly increased total serum IgE level was noted in both atopic (241.2+/- 102.5 IU/mL) and nonatopic children (125.5+/- 63.2 IU/mL) with bronchiolitis compared with that in the control group (27.2+/- 10.5 IU/ml) (P< 0.01). There were significant differences in the proportion of CD4(+)CD25(+) regulatory T cells and Foxp3 mRNA expression in peripheral blood (P< 0.05) as well as total serum IgE level (P< 0.01) between the atopic and the nonatopic group. The proportion of CD4(+)CD25(+) regulatory T cells (r=-0.70, P< 0.01) and Foxp3 mRNA expression in peripheral blood (r=-0.79, P< 0.01) were closely negatively correlated to total serum IgE level.

CONCLUSIONSBoth the proportion of CD4(+)CD25(+) regulatory T cells and Foxp3 mRNA expression in peripheral blood were reduced, in contrast, the total serum IgE level increased in children with RSV bronchiolitis. This suggested that CD4(+)CD25(+) regulatory T cells and Foxp3 mRNA together with IgE participated in the pathogenesis of RSV bronchiolitis.

Bronchiolitis ; etiology ; immunology ; CD4-Positive T-Lymphocytes ; immunology ; Female ; Forkhead Transcription Factors ; genetics ; Humans ; Immunoglobulin E ; blood ; Infant ; Male ; RNA, Messenger ; blood ; Respiratory Syncytial Virus Infections ; etiology ; immunology ; T-Lymphocytes, Regulatory ; physiology

OBJECTIVETo investigate the infection of HEV in Quzhou area of Zhejiang Province.

METHODSAll sera from blood donors in the central blood bank of Quzhou from April 2006 to April 2007 were used. Anti-HEV IgG and anti-HEV IgM were measured by EIA. RT-PCR was also performed to the samples with positive anti-HEV IgM. Genotype and sequence homology were analyzed after sequencing.

RESULTSThe positive ratio of anti-HEV IgG was 40.60%, in which the male infection ratio was higher than the female significantly (43.09% VS 36.09%; chi2=22.6; P < 0.01). The infection ratio was increased with age. The positive ratio of anti-HEV IgM was 0.43%. The positive ratio of anti-HEV IgG and the titers of antibody were higher in the inferior clinical infectors with positive anti-HEV IgM than the negative ones (P < 0.05). Two samples were positive in HEV PCR among 21 samples with positive anti-HEV IgM. The toxemia ratio was 0.4% of all the donors. And the genotype of the two samples with toxemia were both HEV-IV.

CONCLUSIONThe HEV infection was correlation with age and sex significantly and the infection occurred in the adults mainly in Quzhou area. HEV toxemia was not infrequency in the blood donors.

Adolescent ; Adult ; Age Factors ; Antibodies, Viral ; analysis ; immunology ; Blood Donors ; China ; epidemiology ; Female ; Genotype ; Hepatitis E ; complications ; epidemiology ; immunology ; virology ; Hepatitis E virus ; genetics ; immunology ; Humans ; Immunoglobulin G ; analysis ; immunology ; Immunoglobulin M ; analysis ; immunology ; Male ; Middle Aged ; Sequence Analysis, DNA ; Sequence Homology ; Sex Factors ; Toxemia ; complications ; epidemiology ; virology