Asthma ; drug therapy ; immunology ; Diagnosis, Differential ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Major Histocompatibility Complex ; genetics ; immunology ; Medicine, Chinese Traditional ; Phytotherapy

Diisocyanate (DI) is the most common cause of occupational asthma (OA) in Korea. Mannose-binding lectin (MBL) initiates the lectin complement activation pathway following oxidative stress and plays an important role in the regulation of inflammatory processes. To determine whether there is a genetic association between MBL2 polymorphisms and DI-OA, 99 patients with DI-OA, 99 asymptomatic exposed controls (AECs) and 144 unexposed normal controls were enrolled in this study. Three polymorphisms (-554 G>C, - 431A>C and - 225 G>C) in the MBL2 promoter were genotyped, and serum MBL levels were determined by enzyme-linked immunosorbent assay. Functional variabilities in the promoter polymorphisms were analyzed by a luciferase reporter assay and electrophoretic mobility shift assay (EMSA). A significantly higher frequency of haplotype (ht) 2 [CAG] was noted in the DI-OA group compared with the AEC group (P=0.044). The patients with DI-OA carrying ht2 [CAG] had significantly lower PC20 methacholine levels (P<0.001) than the non-carriers. The serum MBL levels were significantly higher in the DI-exposed subjects (both the DI-OA patients and AECs) carrying ht1 [GAG] (P=0.028). Luciferase activity was significantly enhanced in ht1 [GAG] compared with ht2 [CAG] in human hepatocarcinoma cells (Hep3B) (P=0.002). The EMSA showed that a - 554G probe produced a specific shifted band compared with the - 554C probe. These findings suggest that decreased serum MBL levels due to polymorphisms of the MBL2 gene may increase susceptibility to the development of DI-OA in DI-exposed individuals.
Adult ; Alleles ; Asthma, Occupational/diagnosis/*etiology ; Cell Line ; Female ; Forced Expiratory Volume ; Gene Frequency ; Genotype ; Haplotypes ; Humans ; Immunoglobulin E/immunology ; Immunoglobulin G/immunology ; Isocyanates/*adverse effects/immunology ; Male ; Mannose-Binding Lectin/blood/*genetics ; Middle Aged ; *Polymorphism, Genetic ; Polymorphism, Single Nucleotide ; Protein Binding ; Transcriptional Activation ; Young Adult

BACKGROUND: Several methods have been reported to induce asthmatic reactions in mice but few studies have compared their efficiency. We evaluated the efficiency of the protocols frequently used in the literature. METHODS: BALB/c mice were sensitized to ovalbumin (OVA) by intraperitoneal injection; 1] Once a week for two weeks using OVA with alum (IPOA-2) or without (IPO-2), and provocation on days 28-30 by 1% OVA inhalation; 2] seven times for two weeks by OVA with alum (IPOA-7) or without (IPO-7) and provocation by 1% OVA inhalation on days 42-44. 3] Sensitization by 1% OVA inhalation for ten days (IHO-10) and provocation by 1% OVA inhalation on days 28-30. After the last challenge, airway hyperresponsiveness was measured with single chamber plethysmography 24 hours later and mice were sacrificed 48 hours later. RESULTS: Airway hyperresponsiveness, BALF eosinophilia, airway inflammation, and OVA-specific IgE and IgG1 production were effectively induced in IPOA-2, IPOA-7, and IPO-7. However, these phenotypes were not induced in IPO-2 (except for increased BALF eosinophils) or IHO-10 (except for an increased OVA-specific IgG1 level). CONCLUSION: The intraperitoneal injections of OVA with alum once a week for two weeks proved to be the most efficient sensitization method of inducing an asthmatic reaction in mice.
Administration, Inhalation ; Animals ; Antibodies, Anti-Idiotypic/blood ; Asthma/diagnosis/*genetics/immunology ; *Bronchial Provocation Tests ; Comparative Study ; Disease Models, Animal ; Female ; Immunoglobulin G/immunology ; Injections, Intraperitoneal ; Lung/pathology ; Mice ; Mice, Inbred BALB C ; Ovalbumin/administration & dosage/*diagnostic use/immunology ; *Phenotype ; Research Support, Non-U.S. Gov't