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Local Lymph Node Assay
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1. Apply chemical to both ears Days 1,2,3
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2. Inject 3Hthymidine IV Day 6
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3. Remove lymph node and measure proliferation: 5 hour after IV injection
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Figure 19.5 Assessing chemicals for potential contact sensitivity. In the local lymph node assay the chemical in question is applied to both ears on three consecutive days. Control mice are treated with vehicle. Radioisotope is injected intravenously on day 6. The draining lymph nodes are removed 5 hours later and the proliferative response is measured by the incorporation of radio isotope. Results are frequently presented as a stimulation index (counts per min (cpm) for the test chemical/cpm for control). (Picture adapted from D. Sailstad, Lab Animal 31: 36, 2002.)
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contact sensitizers. CHS lends itself to this approach because there is a large database of chemicals known to cause it, and there is a reasonable understanding of chemical characteristics that facilitate skin penetration, chemical reactivity with host proteins, and immune reactivity. Because nonspeci c in ammatory responses also can occur following chemical exposure to the skin, a distinction must be made between an irritant and a sensitizer. An irritant is an agent that causes local in ammatory effects but induces no immunological memory. Therefore, on subsequent exposures, local in ammation will again result, but there is no enhancement of the magnitude of the response and no change in the dose required to induce the response. In immunologically mediated in ammation (hypersensitivity) there may be no response to a sensitizer during the induction stage, but responses to subsequent exposures are exacerbated. The dose required for elicitation is usually less than that required to achieve sensitization.
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19.5.2 Respiratory Allergens
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There is evidence that both occupational and environmental exposures to chemicals (both proteins and haptens) can result in the induction or exacerbation of respiratory allergies (Table 19.6). Of particular concern is the induction of allergic asthma. In sensitized asthmatic individuals the antigen challenge generally causes a type I (IgEmediated) immediate hypersensitivity response with release of mediators responsible for bronchoconstriction. Between 2 and 8 hours after the immediate response, asthmatics experience a more severe and prolonged (late phase) reaction that is characterized by mucus hypersecretion, bronchoconstriction, airway hyperresponsiveness to a variety of nonspeci c stimuli (e.g., histamine, methacholine), and airway in ammation characterized by eosinophils. This later response is not mediated by IgE.
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EFFECTS OF CHEMICALS ON ALLERGIC DISEASE
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Table 19.6 Example of Chemicals Associated with Respiratory Allergy Proteins Enzymes Latex Animal dander Dust mite Molds Cockroach Microbial pesticides Low molecular weight (<3000 )-haptens Toluene diisocyanate Diphenylmethane diisocyanate Phthalic anhydride Trimellitic anhydride Platinum salts Reactive dyes Adjuvants Ozone Nitrogen dioxide Diesel exhaust Residual oil y ash
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Although proteins are generally immunogens, not all proteins are allergens and there is a range of potencies for those that are. There is also a strong genetic component associated with susceptibility to develop allergic reactions to proteins. Susceptible individuals are called atopic. There is at present no structural motif that can be used to characterize a protein as an allergen for hazard identi cation. Examples of occupational protein exposures associated with respiratory allergy and asthma include enzymes, latex, our (both the grain itself and fungal contaminants), and animal dander. Environmental (mostly indoor) exposure including molds, spores, dust mite, animal dander, and cockroach have also been associated with this type of respiratory disease. Because this is a type 1 response, cytophilic antibodies (IgE) speci c for the allergen are frequently used to identify proteins that may cause this effect. For example, in order to determine the etiology of occupational asthma in human subjects, the skin prick test is often used. Different proteins are injected under the skin to test for the presence of cytophilic antibodies in order to identify which proteins are causing a response in an individual. Serum may also be tested for protein speci c IgE. Because IgE can sometimes be detected in the absence of respiratory responses, a positive IgE test may be followed by an assessment of respiratory responses. Under very controlled situations patients may be exposed via the respiratory route to suspect allergens (broncho- provocation test) and respiratory function monitored to pinpoint the offending allergen. Guinea pigs and mice have been used to test proteins for potential allergenicity. Animals are usually sensitized by the respiratory route and monitored for the development of cytophilic antibody (IgG1 in guinea pigs, IgE in mice) as well as increased respiratory rate and other changes in pulmonary function.
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