ovalbumin and anthracene

A reliable and sensitive competitive real-time immuno-PCR (RT-IPCR) assay for the determination of anthracene (AN) was developed. 9-Anthracenebutanoic acid, gamma-oxo-(ANA) was synthesized as the hapten of AN. Mixed anhydride reaction (MAR) was used to couple the ANA to ovalbumin (OVA) to form artificial coating antigen. Active ester method (AEM) was used to couple the ANA to bovine serum albumin (BSA) to form artificial immune antigen. Male New Zealand white rabbits were immunized with immune antigen to obtain polyclonal antibodies (pAbs), with which, a novel RT-IPCR assay for determination of AN was described. Under optimized assay conditions, AN can be determined in the concentration range from 1 fgmL(-1) to 100 pgmL(-1) with a detection limit of 0.5 fgmL(-1). The cross-reactivities of the anti-AN antibody to seven structurally related compounds were below 15%. Environmental water samples were successfully analyzed, showing a good accuracy and suitability to analyze AN in field samples. Recovery was between 93.3% and 120.0% and would be acceptable for use in an on-site field test to provide rapid, semiquantitative, and reliable test results for making environmental decisions.

Topics: Animals; Anthracenes; Antibodies; Antigens; Immunoassay; Male; Ovalbumin; Polymerase Chain Reaction; Rabbits; Reproducibility of Results; Sensitivity and Specificity; Serum Albumin, Bovine; Water

We have previously demonstrated that pyrene in diesel-exhaust particles (DEP) has an adjuvant activity on immunoglobulin E (IgE) antibody production in mice immunized with Japanese cedar pollen allergen (JCPA) or ovalbumin (OA) intraperitoneally. The present study is concerned with the adjuvant activity in IgE antibody production against JCPA of pyrene or DEP inoculated intranasally in mice. We show that anthracene, fluoranthene and benzo(a)pyrene in DEP have the ability to enhance anti-JCPA IgE antibody production in mice by intranasal immunization. Mice were grouped, immunized with 10 micrograms of JCPA plus 400 micrograms of pyrene, 10 micrograms of JCPA plus 100 micrograms of DEP, 10 micrograms of JCPA plus 2 mg of aluminum hydroxide and 10 micrograms of JCPA alone intranasally 7 times at 2 week intervals. Mice were also grouped, and immunized with JCPA (10 micrograms) plus 40 micrograms of anthracene, JCPA (10 micrograms) plus 400 micrograms of fluoranthene, JCPA (10 micrograms) plus 40 micrograms of benzo(a)pyrene, and JCPA (10 micrograms) plus 400 micrograms of pyrene and JCPA (10 micrograms) alone. We found that the IgE antibody responses to JCPA in mice immunized with JCPA plus pyrene, JCPA plus DEP or JCPA plus the three chemical organic compounds mentioned above were significantly enhanced compared with those immunized with JCPA alone. In addition, when the intraperitoneal macrophages obtained from the normal mice (unimmunized mice) were incubated with pyrene, anthracene, fluoranthene or benzo(a)pyrene in vitro, an enhanced chemiluminescence (CI) response and interleukin-1 alpha (IL-1 alpha) production of the macrophages was observed in each instance. These results suggest that in the production of IgE antibody to JCPA the adjuvancy of polycyclic aromatic hydrocarbons (PAHs) in DEP may be important in an attack of Japanese cedar pollinosis.

Topics: Adjuvants, Immunologic; Administration, Intranasal; Air Pollutants; Allergens; Aluminum Hydroxide; Animals; Anthracenes; Antibodies, Anti-Idiotypic; Benzo(a)pyrene; Female; Fluorenes; Food Contamination; Humans; Immunization; Immunoglobulin E; Interleukin-1; Luminescent Measurements; Macrophages, Peritoneal; Male; Mice; Ovalbumin; Pollen; Polycyclic Aromatic Hydrocarbons; Pyrenes; Rats; Rats, Wistar; Rhinitis, Allergic, Seasonal; Trees; Urban Population; Vehicle Emissions