OBJECTIVETo construct a vector encoding T-cell epitopes of major allergen group 1 of Dermatophagoides pteronyssinus as a vaccine delivered by MHC class II pathway.

METHODSThe nucleotide sequences of the 3 target genes were synthesized, including TAT, IhC and the recombinant fragment of Der p 1 encoding 3 T-cell epitopes. After amplification of the 3 target fragments by PCR and digestion with corresponding restriction endonucleases, the recombinant gene TAT-IhC-Der p 1-3T was ligated using T4 DNA ligase and inserted into the prokaryotic expression vector pET28a(+) to construct the recombinant plasmid pET-28a(+)-TAT-IhC-Der p 1-3T, which was confirmed by digestion with restriction endonucleases and sequencing. The recombinant vector was transformed into E. coli strain BL21 (DE3) and induced with IPTG, and the induced protein TAT-IhC-Der p 1-3T was detected by SDS-PAGE. After purification, the recombinant protein was confirmed by Western blotting and its allergenicity tested using IgE-binding assay.

RESULTSThe recombinant plasmid pET-28a-TAT-IhC-Der p 1-3T was successfully constructed as confirmed by restriction endonuclease digestion and sequencing and the expression of the recombinant protein TAT-IhC-Der p 1-3T was induced in E. coli. Western blotting verified successfull purification of the target protein, which showed a stronger IgE-binding ability than Der p 1.

CONCLUSIONWe successfully constructed a recombinant expression vector pET-28a-TAT-IhC-Der p 1-3T expressing a T-cell epitope vaccine delivered by MHC II pathway with strong IgE-binding ability, which provides a basis for further study on specific immunotherapy via MHC class II pathway.

Allergens ; immunology ; Animals ; Antigens, Dermatophagoides ; immunology ; Arthropod Proteins ; immunology ; Base Sequence ; Cloning, Molecular ; Cysteine Endopeptidases ; immunology ; Dermatophagoides pteronyssinus ; Epitopes, T-Lymphocyte ; Escherichia coli ; Gene Expression ; Genes, MHC Class II ; Genetic Vectors ; Plasmids ; Polymerase Chain Reaction ; Recombinant Proteins ; immunology ; Vaccines ; immunology

OBJECTIVE: To evaluate the efficacy and safety of the sublingual immunotherapy with dermatophagoides farinae drops on patients with allergic rhinitis. METHOD: One hundred and twelve cases were collected from adult patients with dust-mite allergic rhinitis of our hospital who could adhere to treatment and regular follow-up. These patients were randomly allocated to receive either sublingual immunotherapy (SLIT group, n = 56) or medical treatment (Control group, n = 56). To evaluate the clinical efficacy by side effects which were registered, symptom and medication scores which were assessed and rhinoconjunctivitis quality of life questionnaire (RQLQ) which was completed in the baseline and two years after treatment. RESULT: Dropouts after the 2 years' treatment were 5 of SLIT group and 4 of Control group respectively. SLIT group induced the significant reductions on both the symptom scores (7.81 ± 3.14 to 3.89 ± 2.01, P < 0.0 1) and the medication scores (2.86 ± 0.75 to 0.44 ± 0.06, P < 0.01). Meanwhile, Control group induced the reductions on both the symptom scores (8.01 ± 3.32 to 5.20 ± 2.43) and the medication scores (2.95 ± 0.80 to 1.75 ± 0.40). There were significant differences (P< 0. 01) in symptom and medication scores between the two groups after 2-year treatment. The patients in SLIT group had fewer symptoms and lower intake of medication. There were statistically significant differences in RQLQ between SLIT group [19 (15,22)] and Control group [36 (26,47)] after two years treatment (Z = -5. 21, P < 0.01). SLIT group also had significant improvement in RQLQ (Z = -6.10, P < 0.01) between before and after the treatment. There were 4 patients who showed adverse reactions in SLIT group (3 occurred in increment period, and 1 occurred in the maintenance period). The incidence of adverse reactions was 7.14%. No severe systemic side effects were registered. CONCLUSION SLIT with standardized dermatophagoides farinae drops in China is safe and effective to patients with allergic rhinitis.
Administration, Sublingual ; Adult ; Animals ; Antigens, Dermatophagoides ; immunology ; China ; Dermatophagoides farinae ; Humans ; Quality of Life ; Rhinitis, Allergic ; drug therapy ; Sublingual Immunotherapy ; Treatment Outcome

OBJECTIVEThe prevalence of allergic asthma has been rising continually which is correlated with the increasingly closed living environment. House dust mites are the major sources of indoor aeroallergens which induce asthma in sensitized people. To monitor the seasonal variation of house dust mite (HDM)-allergens exposure level in the asthmatic children, which was evaluated to show its correlation with the level of asthma control, HDM allergic sensitization and fraction of exhaled nitric oxide, and to provide basic data for HDM environmental control.

METHODA total of 48 HDM-allergic asthmatic children were enrolled from the asthma clinic in the hospital from March 2011 to January 2012 (boys 34 and girls 14, aging from 3 to 14 years, mean age 8 years and 4 months) in the present study. Dust samples from mattresses, pillows, bedroom floor, living room floor and sofas were collected in each season within one year in the household of all the enrolled patients. The concentrations of Der p 1 and Der f 1 were measured by the enzyme-linked immunosorbent assay (ELISA). To record the Asthma Control Test (ACT) score or Children Asthma Control Test (C-ACT) score for each patient and to collect the data of doctor monitoring asthma control level each time when the patient was clinic visited. The concentration and its classification of the serum specific IgE to HDM was determined by fluoroenzyme-immunometric assay.

RESULTThe average concentration of Der f 1 of all dust samples was significantly higher than that of Der p 1 (0.13 µg/g vs 0.02 µg/g, P < 0.05). The concentrations of Der f 1 from mattresses, pillows and sofas dust samples were significantly higher than those from bedroom floor and living room floor dust samples (0.69 µg/g, 0.42 µg/g and 0.22 µg/g vs 0.07 µg/g and 0.07 µg/g, P < 0.05). The Der f 1 exposure level from mattress dusts in summer but no others was negatively correlated with asthma control level (r = -0.318, P = 0.036). The Der f 1 exposure level from any area dusts in summer and the Der p 1 exposure level from pillows dusts in autumn was negatively correlated with ACT/C-ACT score respectively. The Der f 1 from mattress dusts in winter was positively correlated with classification of sIgE to Der f 1. The Der p 1 exposure level from most areas in each season was positively correlated with classification of sIgE to Der f 1 and Der P 1.

CONCLUSIONDer f 1 was the predominant mite allergen in household dust and mainly came from mattresses, pillows and sofas. The role of the HDM allergen exposure level on the asthma control level and HDM allergic sensitization for the asthmatic children were depended on its area, season and HDM species, which suggested that the future assessment of clinical effect by the HDM environmental control should consider these factors.

Adolescent ; Air Pollution, Indoor ; adverse effects ; Animals ; Antigens, Dermatophagoides ; analysis ; Asthma ; diagnosis ; etiology ; immunology ; Child ; Child, Preschool ; Dust ; Environmental Exposure ; Enzyme-Linked Immunosorbent Assay ; Female ; Forced Expiratory Volume ; Humans ; Immunoglobulin E ; blood ; Male ; Pyroglyphidae ; immunology ; Risk Factors ; Seasons

OBJECTIVE: To compare the results of skin prick test with allergens of Allergopharma company or ALK company in Shanghai area. METHOD: The study included 9 233 patients diagnosed as allergic rhinitis. The patients were divided in to two groups (Allergopharma group and Alutard-SQ group) SPT was given from June, 2005 to Feb, 2013. The results of the positive rate using two reagents were analyzed and compared. RESULT: There were no significant differences between the two groups in age and gender (P > 0.05). The total positive rate in Allergopharma group was 62.90%, and 59.68% in Alutard-SQ group. The positive rate in Allergopharma group was significantly higher than that in Alutard-SQ group (P < 0.01). Dermatophagoides farinae (Der f) and Dermatophagoides pteronyssinus (Der p) were identified as the mostly responsible allergens. The positive rate of Der f in Allergopharma group and Alutard-SQ group was 55.23% and 45.57% respectively. The same rate were 54.83% and 47.89% of Der p. There were significant differences of the positive rate of two allergens in both groups (P < 0.01). CONCLUSION The total positive rate was different using different allergens for SPT among the patients diagnosed as allergic rhinitis of the same area. Dust mites were identified as main allergens. The positive rate was different when using different reagents, which needed more attention to make the decision of specific immune therapy.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Allergens ; immunology ; Antigens, Dermatophagoides ; immunology ; Child ; Child, Preschool ; China ; Female ; Humans ; Indicators and Reagents ; Male ; Middle Aged ; Rhinitis, Allergic ; diagnosis ; Skin Tests ; Young Adult

OBJECTIVETo evaluate the efficacy of specific sublingual immunotherapy (SLIT) with Dermatophagoides farinae drops on children with allergic asthma and allergic rhinitis of the preschool and school-age groups of children and adolescents.

METHODThis study analyzed the efficacy of SLIT in 122 children (aged 3-14 yr) with house dust mites-induced allergic asthma and allergic rhinitis. According to the age, patients were defined as the preschool group ( ≤ 6 years old, n = 59) and school-age group (> 6 years old, n = 63). All children were treated with Dermatophagoides farinae drops for at least 2 years. Clinical observation and follow-up study were conducted during the treatment. Before and after SLIT for half a year, 1 year and 2 years, asthma symptom scores (day and night), rhinitis symptom scores and medication scores were evaluated. The adverse events were assessed during the study.

RESULTAfter SLIT for half a year, 1 year and 2 years, there were no significant differences between the preschool group (0.3 ± 0.5,0.0 ± 0.1,0.0 ± 0.0) and school-age group (0.3 ± 0.4,0.0 ± 0.1,0.0 ± 0.0) in day scores of asthma (Z value was -1.687, -0.613,0.000, all P > 0.05). There were no significant differences between the preschool group (0.2 ± 0.5,0.1 ± 0.3,0.0 ± 0.0) and school-age group (0.2 ± 0.4,0.1 ± 0.3, 0.0 ± 0.0) in night scores of asthma (Z value was -0.496, -0.486,0.000, all P > 0.05). There was no significant differences between the preschool group (1.4 ± 0.9,0.4 ± 0.5,0.1 ± 0.3) and school-age group (1.3 ± 0.9,0.5 ± 0.6,0.2 ± 0.4) in symptom scores of allergic rhinitis (Z value was -0.394, -1.166, -1.075, all P > 0.05). There were no significant differences between the preschool group (1.6 ± 0.8,0.0 ± 0.0,0.0 ± 0.0) and school-age group (1.7 ± 0.7,0.0 ± 0.0,0.0 ± 0.0) in medication scores of allergic rhinitis (Z value was -0.655,0.000,0.000, all P > 0.05). After SLIT for 2 years, most children in the preschool and school-age groups were no longer using asthma controlling medication, with no significant difference between the two groups (Z value was 0.000, P > 0.05). The overall incidence of adverse reactions was only 7%, and there was no significant difference in the incidence of adverse reactions between the two groups (χ(2) = 0.000, P > 0.05). The local adverse events were mild gastrointestinal discomfort and rash, a majority of local adverse events in the preschool group were diarrhea. No severe adverse events happened in the treatment.

CONCLUSIONSLIT with Dermatophagoides farinae drops is safe and effective in children with allergic asthma and allergic rhinitis of the preschool and school-age groups of children and adolescents, which provides evidences for early SLIT intervention of the disease.

Administration, Sublingual ; Adolescent ; Antigens, Dermatophagoides ; administration & dosage ; immunology ; Asthma ; immunology ; therapy ; Child ; Child, Preschool ; Female ; Follow-Up Studies ; Humans ; Male ; Rhinitis, Allergic, Perennial ; immunology ; therapy ; Sublingual Immunotherapy ; Treatment Outcome

OBJECTIVETo observe the efficacy of sublingual immunotherapy (SLIT) in children with allergic asthma during the treatment and 1 year after the treatment.

METHODThis is an open and retrospective study; 80 children with mild-moderate allergic asthma between 4 and 14 years of age were chosen from the Department of Respiratory Medicine, Nanjing Children's Hospital Affiliated to Nanjing Medical University from May to August, 2009. All children were sensitized to Dermatophagoides Farianae and/or Dermatophagoides Pteronyssinus and have received anti-asthma drug therapy for 3 months (baseline). Thirty-nine children in SLIT group underwent 2-year SLIT and combined with anti-asthma drug, these children were then followed up for 1 year. Forty-one children in drug group only received anti-asthma drug and were followed up for 3 years. The scores of asthma symptom, scores of asthma medication and the number of discontinuation of anti-asthma drug were compared between the SLIT group and drug group for the baseline, end of the 2nd year and 3rd year treatment. The frequency of acute attack of asthma was also compared between the two groups for 1 year before the treatment and the 3rd year treatment.

RESULT(1) At baseline, the asthma symptom scores, the medication scores and the frequency of acute attack of asthma in 1 year before the treatment of the two groups showed no significant difference. (2) After 2-year SLIT, the daytime asthma symptom scores of SLIT group were lower than the drug group (0.18 ± 0.06,0.93 ± 0.12,Z = -4.873, P < 0.05), the night asthma symptom scores of the two groups showed no significant difference. One year after SLIT, the daytime and night asthma symptom scores of SLIT group were both lower than those of the drug group (daytime SLIT group vs. Drug group: 0.18 ± 0.06 vs. 1.46 ± 0.72,Z = -5.082, P < 0.05;night SLIT group vs. Drug group: 0.05 ± 0.04 vs. 0.66 ± 0.14,Z = -4.019, P < 0.05). (3) At the end of SLIT and 1 year after SLIT, the medication scores of SLIT group were both lower than those of the drug group (End of SLIT SLIT group vs. Drug group: 0.31 ± 0.07 vs. 0.75 ± 0.12,Z = -2.813, P < 0.05;1 year after SLIT SLIT group vs. Drug group: 0.17 ± 0.06 vs. 0.87 ± 0.17,Z = -4.106, P < 0.05), the number of discontinuation of anti-asthma drug of SLIT group were both more than the drug group (End of SLIT SLIT group vs. Drug group: 20 vs. 10,χ(2) = 6.167, P < 0.05;1 year after SLIT SLIT group vs. Drug group: 29 vs.13,χ(2) = 14.581, P < 0.05).(4) In the 3rd year, the frequency of acute attack of asthma in SLIT group was significantly lower than that of drug group (0.69 ± 1.20, 1.20 ± 1.44,Z = -1.968, P < 0.05) .

CONCLUSIONSLIT can significantly improve the symptoms of asthma, reduce the use of anti-asthma drug and reduce the frequency of the acute attack of asthma. Meanwhile, the efficacy could still maintain 1 year after the SLIT treatment.

Administration, Sublingual ; Adolescent ; Animals ; Anti-Asthmatic Agents ; therapeutic use ; Antigens, Dermatophagoides ; administration & dosage ; immunology ; Asthma ; drug therapy ; immunology ; therapy ; Case-Control Studies ; Child ; Female ; Follow-Up Studies ; Humans ; Male ; Pyroglyphidae ; immunology ; Retrospective Studies ; Sublingual Immunotherapy ; Treatment Outcome

We attempted to investigate the correlation between the severity of atopic dermatitis (AD) in children and the indoor level of house dust mite (HDM) allergens. Ninety-five patients (31.1 +/- 19.5 months of age) with AD were enrolled in this study, and serum specific IgE against Dermatophagoides pteronyssinus and D. farinae was measured. The severity of AD was assessed using the visual analogue scale on the same day of house dust collection. Living rooms and mattresses where the child usually slept were vacuumed for 2 minutes and concentrations of Der f 1 were measured by enzyme-linked immunosorbent assay. The skin symptoms were more severe in patients with Der f 1 concentrations in living room > 2 microg/g dust than < or = 2 microg/g dust (P = 0.018). This difference was noted in AD patients without sensitization to HDM (P = 0.004), but not in patients with sensitization. There was no difference in symptom severity according to Der f 1 concentrations in mattresses (P = 0.062). The severity of skin symptoms is associated with indoor concentrations of HDM in children with AD, and it is likely to act as nonspecific irritants as well as allergens in AD skin lesions.
Adolescent ; Animals ; Antigens, Dermatophagoides/*analysis ; Beds/parasitology ; Child ; Child, Preschool ; Dermatitis, Atopic/*diagnosis/pathology ; Dermatophagoides farinae/immunology ; Dermatophagoides pteronyssinus/immunology ; *Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Immunoglobulin E/blood ; Infant ; Male ; Severity of Illness Index

OBJECTIVE: To evaluate the efficacy and safety of mite allergen specific immunotherapy (SIT) in treating children with allergic asthma. METHOD: A total of 136 patients with mite allergy were recruited into the study. They were randomly divided into two groups: SIT group (n = 66) and ST (symptomatic therapy) group (n = 70). They were investigated of SIT with standardized allergen vaccine or no SIT only symptomatic therapy respectively. Therapeutic evaluation index includes: asthma symptoms score, drug score, skin prick test, pulmonary function, serum specificity IgE (sIgE) and the new sensitization was also assessed. Local and systemic adverse reactions were used to evaluate the clinical safety. RESULT: Clinical symptom scores, drug scores, Lung function, and skin test result all improved significantly after the treatment with SIT compared to ST group (P < 0.01). SIT groups do not have new sensitization and no fatal systemic reactions occurred. CONCLUSION The standardized dust mite allergen specific immunotherapy is efficacious and safe to Children with allergic asthma . SIT can reduce house dust mites skin sensitivity and prevent new allergen appeared.
Adolescent ; Allergens ; therapeutic use ; Animals ; Antigens, Dermatophagoides ; therapeutic use ; Asthma ; immunology ; therapy ; Child ; Dust ; Female ; Humans ; Hypersensitivity ; immunology ; therapy ; Immunotherapy ; methods ; Male ; Pyroglyphidae ; immunology ; Safety ; Sensitivity and Specificity ; Skin Tests ; Vaccines ; therapeutic use

OBJECTIVE: To investigate the correlation of serum specific IgE and skin prick test in allergic rhinitis patients sensitive to dust mite and the difference between the results. METHOD: Data of 349 allergic rhinitis patients who had positive result in either Dermatophagoides pteronyssinus(Der p) or Dermatophagoides farinae (Der f) by serum specific IgE and skin prick test were statistically analyzed. RESULT: Grades of Skin prick test and specific IgE levels were notably relevant in these two dust mites(Der p r= 0. 568, Der f r= 0. 506, P<0. 01). There was significant difference of positive rates between serum specific IgE and skin prick test in Der p(Chi2 = 11. 605, P<0. 01)but not in Der f(Chi2 =0, P>0.05). There was no correlation between positive degree of two methods and score of clinical symptoms in allergic rhinitis. CONCLUSION Serum specific IgE and skin prick test were notably relevant in allergic rhinitis patients sensitive to dust mites. The positive rates of the two methods were different due to different allergen. Therefore, they could not substitute for each other. The level of serum specific IgE and positive degree of skin prick test could not reflect the degree of symptom in allergic rhinitis.
Adolescent ; Adult ; Aged ; Animals ; Antigens, Dermatophagoides ; immunology ; Child ; Child, Preschool ; Female ; Humans ; Immunoglobulin E ; blood ; Male ; Middle Aged ; Pyroglyphidae ; immunology ; Rhinitis, Allergic ; Rhinitis, Allergic, Perennial ; blood ; diagnosis ; immunology ; Skin Tests ; Young Adult

OBJECTIVETo investigate the incidence of local reactions (LRs) and systemic reactions (SRs) of subcutaneous immunotherapy (SCIT) and to analyze the potential risk factors of such reactions in Chinese population.

METHODThis is a retrospective study on 234 dust mite sensitized patients with allergic rhinitis and asthma who received allergen immunotherapy in our hospital from 2003 to 2010. Chart review was conducted to capture clinical data of reactions to immunotherapy. Parameters included signs and symptoms, the onset of reaction, and interventions in treating such reactions, particularly, the administration of epinephrine (EPI) and adjustment of vaccine dosage due to LRs and SRs.

RESULTThe 234 patients received a total of 7679 injections. Among them, 4973 LRs (64.8%) and 235 SRs (3.1%) were observed in 67 patients (28.6% of all patients). SRs included respiratory symptoms (205 events, 88.4%) and cutaneous symptoms (31.5%). Of the total of 235 SR events, 212 (90.2%) were presented as mild SRs and 23 (9.8%) were in severe SR category (grade III and grade IV, EAACI grading system). Overall, severe SRs accounted for 0.3% of total injections. Seventeen of the 23 SR events required epinephrine treatment (0.2% of total injections). Of the 67 patients, 61 completed the course of treatment after dose adjustment; 36 patients had their doses decreased prior to further advancing to target dose. Nineteen subjects tolerated splitting two injections at 30 minutes interval. Six patients advanced the dose based on protocol and another 6 had to stop immunotherapy. Most of the SRs (77.4%) occurred during the maintenance phase of immunotherapy. The levels of TIgE, SIgE D1 and SIgE D2 were found to be significantly higher in patients with SRs comparing to patients without SRs (P < 0.05). SRs more commonly occurred in patients with age less than 14 years than their older counterparts (95.5% vs. 85.6%, OR = 3.58, 95%CI = 1.040 - 12.322, P < 0.01). The incidence of SRs were significantly higher in asthma patients who received SCIT than non-asthma patients (OR = 2, 95%CI = 1.136 - 4.624).

CONCLUSIONOur study suggests that risk factors of SRs include maintenance phase (higher allergen vaccine doses), patients with asthma, age of less than 14 years, higher levels of TIgE, and SIgE D1 and SIgE D2. Effective management includes proper dose adjustment, splitting doses into 2 injections at 30 min apart, and strictly following immunotherapy indications.

Adolescent ; Adult ; Animals ; Antigens, Dermatophagoides ; administration & dosage ; immunology ; Asthma ; immunology ; therapy ; Child ; Child, Preschool ; Desensitization, Immunologic ; adverse effects ; methods ; Female ; Humans ; Hypersensitivity, Immediate ; epidemiology ; etiology ; therapy ; Injections, Subcutaneous ; Male ; Middle Aged ; Mites ; immunology ; Retrospective Studies ; Rhinitis, Allergic, Perennial ; immunology ; therapy ; Risk Assessment ; Treatment Outcome ; Young Adult