OBJECTIVETo identify the IgE-binding epitopes in the allergen Der p 1 of main house dust mites, which can be recognized by the specific IgE in the sera from allergic individuals, and obtain a hypoallergen derived from the T-B epitope fused peptide for potential use in specific immunotherapy (SIT).

METHODSThirty-one peptides containing 15 amino acids each, which covered the full 222 amino acids of Der p 1 protein sequence, were synthesized on the cellulous membrane by solid-phase peptide (SPOTs) synthesis, with 8 overlapping amino acids between every two neighboring peptides. The membrane bearing the spots of the synthesized peptides were incubated with the allergic serum pools consisting of the sera from 5 allergic individuals. The membrane was then probed with HRP-conjugated anti-human IgE, followed by enhanced chemiluminescence (ECL) for visualization and gray scale analysis of the positive peptide spots.

RESULTSThree strong IgE-binding epitopes were identified in the amino acid sequence of Der p 1 molecule, namely Ep1 (amino acids 85-99), Ep2 (amino acids 106-120) and Ep3 (amino acids 190-204).

CONCLUSIONThe 3 IgE-binding epitopes (B cell epitopes) identified in Der p 1 confirm the presence of linear epitopes in Der p 1, suggesting the possibility of constructing T/B epitope-fused hypoallergens.

Amino Acid Sequence ; Animals ; Antigens, Dermatophagoides ; immunology ; Arthropod Proteins ; immunology ; Binding Sites, Antibody ; Cysteine Endopeptidases ; immunology ; Epitopes ; immunology ; Immunoglobulin E ; immunology ; Lymphokines ; immunology ; Mites ; immunology ; Molecular Sequence Data

OBJECTIVE: To investigate aeroallergen spectrum and allergy positive rates of patients with child allergic rhinitis and analyze its related factors. METHOD: Skin prick test was carried out in 562 cases with child allergic rhinitis using 13 inhaled allergens, and detailed history was collected in all cases. RESULT: Four hundred and fifty-seven (81.3%) of 562 cases showed positive reaction to at least one allergen out of 13 allergens. The most common allergens found in patients was Dermatophagoides farinae and Dermatophagoides pteronyssinus, and positive reaction was 93.1% in child allergic rhinitis, followed by tropical mite, Periplaneta americana, Blattella germanica and dog hair. The pollen allergen, most common in American and European, was pretty rare in this study. The prevalence of child allergic rhinitis was significantly higher in urban than in rural (P<0.01). There was no adverse effect appeared in children with allergic rhinitis underwent skin prick test. CONCLUSION The major allergen of allergic rhinitis is mite for child allergic rhinitis, and relates to housing enviroment. Most of patients with child allergic rhinitis can be treated by the mite specific immunotherapy. The skin prick test is a safe technique for diagnosis of children with allergic rhinitis.
Allergens ; immunology ; Animals ; Asthma ; epidemiology ; immunology ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Hypersensitivity, Immediate ; immunology ; Male ; Mites ; immunology ; Rhinitis, Allergic, Perennial ; epidemiology ; immunology ; Skin Tests

No abstract available.
Animal ; Antibodies, Bacterial/blood ; Antigens, Bacterial/blood ; Human ; Mites/microbiology ; Orientia tsutsugamushi/classification/immunology ; Scrub Typhus/diagnosis/*epidemiology/immunology

Higher levels of IgE-containing immune complexes (IC) have been reported in sera from patients with allergic diseases than in sera from controls. To evaluate the possibility of an IC-mediated mechanism in the pathogenesis of bronchial asthma, we measured circulating C3-containing IgE IC (C3-IgE IC) using anti-C3 ELISA from 20 house dust mite (HDM)-sensitive asthmatics, 20 non-atopic asthmatics, and 14 non-atopic controls. C3-IgE IC levels were significantly higher in HDM-sensitive asthmatics (mean +/- S.D.: 12.2 +/- 7.8 AU/ml) than in non-atopic asthmatics (6.5 +/- 7.5 AU/ml) or controls (5.8 +/- 4.4 AU/ml). C3-IgE IC levels were significantly correlated with HDM-specific IgE levels (r = 0.50, p<0.05), but not with total IgE levels (r = 0.36, p< 0.05) in HDM-sensitive atopic asthmatics. C3-IgE IC levels in sera did not significantly change during HDM-bronchoprovocation test in six HDM-sensitive asthmatics who showed positive reaction. Part of C3-IgE IC could be precipitated by protein G coupled beads. In conclusion, C3-IgE IC levels were elevated in sera from HDM-sensitive asthmatics; moreover IgG antibodies might be a component of C3-IgE IC. Our results suggest that an IgE IC-mediated mechanism could be involved in the pathogenesis of atopic asthma.
Adult ; Animal ; Antigen-Antibody Complex/*blood ; Asthma/*immunology ; Complement 3/*analysis ; Dust ; Human ; Immunoglobulin E/*blood ; Mites/immunology

OBJECTIVE: To analyze the results of serum in allergic rhinitis and investigate the specific clinic allergen and serum immunoglobulin E (IgE) levels. METHOD: Allergy Screen method was used to detect the specific allergen and total serum IgE level of 134 cases of Allergic rhinitis. RESULT: The dust mite was the most common allergen in inhalation group in 134 cases of allergic rhinitis, the positive rates was 90%; then were donly, feline and scurfy fungus, the positive rates were 16%, 9%. The positive rates of total IgE was 54%. The serum IgE levels between 100 to 200 kU/L, there was 21 cases together, but there existed 7 negative cases. There were 51 cases' IgE levels more than 200 kU/L, the rates was 70.8%, but there still existed 4 negative cases. CONCLUSION Allergy screen method can find relevant allergen and provide basis for the prevention and treatment of allergic disease.
Adult ; Allergens ; blood ; immunology ; Animals ; Antigens, Dermatophagoides ; analysis ; Female ; Humans ; Immunoglobulin E ; blood ; Male ; Mites ; immunology ; Passive Cutaneous Anaphylaxis ; Pollen ; immunology ; Rhinitis, Allergic, Perennial ; diagnosis ; immunology

The allergens were separated from the extracts of house dust mites by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and identified by autoradiography. Over 30 protein bands of the whole body extract of Dermatophagoides farinae were apparent on 10-20% gradient SDS-PAGE, and 13 bands with MW between 93KD and 12KD bound with specific IgE antibodies in patients' sera sensitive to house dust mites. The major allergenic component of the whole body extract of D. farinae was the protein of MW 14-15KD, which was detected in 95.7% of 47 patients' sera sensitive to house dust mites. The extract of Dermatophagoides pteronyssinus supplied by Bencard Company, England was thought to contain feces enriched material as noted in a few broad protein bands on SDS-PAGE. Seven allergenic components were shown by autoradiography. The protein band of MW 14-15KD was one of the most frequently revealed allergens on autoradiography, which has appeared in 32.5% of 40 patients' sera sensitive to house dust mites. The electrobotting technique used in the present study was fast, convenient and highly useful for both the identification of allergen components and the screening of specific IgE antibody. The individual variations of IgE immune responses to the allergenic components of the two house dust mites were discussed.
Allergens/*analysis ; Animals ; Autoradiography ; Dust ; Electrophoresis, Polyacrylamide Gel ; Human ; Immunoglobulin E/metabolism ; Mites/*immunology ; Support, Non-U.S. Gov't

OBJECTIVE: To investigate the allergen distribution of subjects suffered from the allergic rhinitis in Changji district. METHOD: Skin prick test was employed on all the 1 564 sufferers by 17 sorts of allergic stock solution, with the physiological saline to be the negative control and the histamine to be the positive control. RESULT: The positive rate of allergen citanest was 85.17%. Furthermore, Chenopodium and Lupulus, as the main allergen substances, were found out to be the highest positive rate among 1332 subjects who were detected to be positive, with pollen of trees following. Whereas, the positive rate of allergic rhinitis caused by dust mite, fungus and canine epidermis was relatively low. CONCLUSION The main allergens of the allergic rhinitis in Changji district are Chenopodium and Lupulus, similar as the dust mite. The allergen prick test, which is characterized as accuracy, sensitiveness and fastness, is an essential way to seek and screen those allergens. Thus, it can provide scientific instruction to the prevention of the allergic rhinitis in Changji district.
Adolescent ; Adult ; Aged ; Allergens ; immunology ; Animals ; Asthma ; etiology ; immunology ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Fungi ; immunology ; Humans ; Male ; Middle Aged ; Mites ; immunology ; Pollen ; immunology ; Rhinitis, Allergic, Perennial ; epidemiology ; immunology ; Skin Tests ; Young Adult

OBJECTIVE: To explore the influence of sublingual immunotherapy on Th2 type immunoreaction in patients with allergic rhinitis within 12 months' treatment. METHOD: Forty three patients with allergic rhinitis were followed-up within 12 months after receiving sublingual immunotherapy. The changes of clinic symptoms and immunoreaction were analyzed. VAS score was applied to evaluate the efficacy of clinic treatment. TIgE and sIgE in serum were tested to evaluate the systemic immunoreaction. Eosinophil counting in nasal discharge was applied to evaluate the local nasal mucosa immunoreaction. RESULT: VAS scores were decreased significantly one month after the starting point of treatment, companied by the decrease of EOS count in nasal discharge (P < 0.05). The changes of TIgE and sIgE in serum within 12 months of sublingual immunotherapy were not significant (P < 0.05). The levels of IL-4 and IL-13 in nasal lavage were decreased significantly after 12 months' treatment. CONCLUSION The bias of Th2 type immunoreaction existed in nasal mucosa immune system in patients with allergic rhinitis could been rectified partially after receiving sublingual immunotherapy for 12 months, but the significant changes were not observed in systemic immune system.
Adolescent ; Adult ; Animals ; Child ; Eosinophils ; immunology ; Female ; Humans ; Immunoglobulin E ; blood ; Interleukin-13 ; immunology ; Interleukin-4 ; immunology ; Male ; Middle Aged ; Mites ; immunology ; Rhinitis, Allergic ; immunology ; therapy ; Sublingual Immunotherapy ; Th2 Cells ; immunology ; Young Adult

The major house-dust-mite allergen, Der p I, stimulates the phospholipase D (PLD) in peripheral blood mononuclear cells (PBMC) from allergic patients with maximal responses after 30 min exposure. At 30 min, Der p I stimulated PLD activity by 1.4-fold in mild, 1.6-fold in moderate and 2-fold in severe allergic patients over control values (p < 0.05). When the cells were pretreated for 24 h with phorbol myristate acetate to down-regulate protein kinase C (PKC), PLD stimulation by Der p I was largely abolished. These results indicate that in PBMC from allergic patients, Der p I can stimulate PLD activity, and that PKC activation is involved in this stimulation.
Adult ; Allergens/metabolism* ; Allergens/immunology ; Animal ; Down-Regulation (Physiology) ; Glycoproteins/metabolism* ; Glycoproteins/immunology ; Human ; Hypersensitivity/metabolism ; Hypersensitivity/immunology ; Hypersensitivity/blood ; IgE/blood ; In Vitro ; Leukocytes, Mononuclear/metabolism ; Leukocytes, Mononuclear/immunology ; Mites/metabolism ; Mites/immunology ; Phospholipase D/metabolism* ; Phospholipase D/immunology ; Protein Kinase C/metabolism* ; Skin Tests ; Tetradecanoylphorbol Acetate/pharmacology

To evaluate the in vivo effect of autologous serum including antibodies to house dust mite in atopic individuals, we observed the immediate (15 mins) and late (6 hours) skin reactions (ISR, LSR) on intradermal (ID) test of serially diluted Dermatophagoides farinae antigens (DFa, Allergopharma, Germany) mixed with autologous sera (DFa-S) and diluent alone (DFa-D). We tested 34 DFa-skin reactive atopic individuals including 12 asthmatics (BA), 8 asthmatics on immunotherapy with DFa (IT), and 14 healthy atopic controls (AC). We observed complete inhibition of ISR in the lowest allergen dose of DFa-S in 7 (58.3%) of 12 BA, 3 (37.5%) of 8 IT, and 2 (14.3%) of 14 AC. In BA, the inhibition of ISR was more frequent than AC (p< 0.05). We observed larger late reactions in half of LSR positive cases on ID test by DFa-S than by DFa-D (> or = 1.5 X size; accentuation of LSR). Accentuation of LSR were shown more frequently by DFa mixed with larger amount of serum (25% in 1:1 mix; 80% in 1:3 mix, p< 0.05). But there were no differences of DFa-specific IgE and IgG subclass antibodies regardless of the inhibition of ISR or the accentuation of LSR. In conclusion, some autologous sera from DFa-sensitive individuals showed the inhibition of ISR and the accentuation of LSR on DFa-ID test.
Animal ; *Blood Physiology ; Dermatitis, Atopic/blood/*immunology ; Human ; Hypersensitivity, Delayed/*immunology ; Hypersensitivity, Immediate/*immunology ; Immunoglobulin E/metabolism ; Immunoglobulin G/metabolism ; Intradermal Tests ; Mites/*immunology ; Skin/*immunology ; Support, Non-U.S. Gov't