Laboratory animal models have been developed to investigate preventive or therapeutic effect of medicinal products, or occurrence or progression mechanism of atopic dermatitis (AD), a pruritic and persistent inflammatory skin disease. The murine model with immunologic phenomena resembling human AD was introduced, which demonstrated skewedness toward predominance of type-2 helper T cell reactivity and pathophysiological changes similar as human AD following 2,4-dinitrochlorobenzene (DNCB) sensitization and challenge. Molecular mechanism on the DNCB-mediated AD was further evaluated. Skin tissues were collected from mice treated with DNCB, and each tissue was equally divided into two sections; one for protein and the other for mRNA analysis. Expression of filaggrin, an important protein for keratinocyte integrity, was evaluated through SDS-PAGE. Level of mRNA expression for cytokines was determined through semi-quantitative reverse transcriptase polymerase chain reaction. Expression of filaggrin protein was significantly enhanced in the mice treated with DNCB compared with the vehicle (acetone : olive oil = 4 : 1 mixture) treatment group or the normal group without any treatment. Level of tumor necrosis factor-alpha and interleukin-18 mRNA expression, cytokines involved in activity of type-1 helper T (TH1) cell, was significantly downregulated in the AD group compared with other control groups. These results suggest that suppression of TH1 cell-mediated immune response could be reflected into the skin tissue of mice treated with DNCB for AD induction, and disturbance of keratinocyte integrity might evoke a compensatory mechanism.
In commercial products such as household deodorants or biocides, didecyldimethylammonium chloride (DDAC) often serves as an antimicrobial agent, citral serves as a fragrance agent, and the excipient ethylene glycol (EG) is used to dissolve the active ingredients. The skin sensitization (SS) potentials of each of these substances are still being debated. Moreover, mixtures of DDAC or citral with EG have not been evaluated for SS potency. The in vitro alternative assay called human Cell Line Activation Test (h-CLAT) and Direct Peptide Reactivity Assay (DPRA) served to address these issues. On three independent runs of h-CLAT, DDAC and citral were predicted to be sensitizers while EG was predicted to be a non-sensitizer and also by the DPRA. Mixtures of DDAC or citral with EG at ratios of 7:3 and 1:4 w/v were all positive by the h-CLAT in terms of SS potential but SS potency was mitigated as the proportion of EG increased. Citral and its EG mixtures were all positive but DDAC and its EG mixtures were all negative by the DPRA, indicating that the DPRA method is not suitable for chemicals with pro-hapten characteristics. Since humans can be occupationally or environmentally exposed to mixtures of excipients with active ingredients, the present study may give insights into further investigations of the SS potentials of various chemical mixtures.
Exposure to bioaerosols in indoor animal farms associates with respiratory illnesses, but little is known about the immune modulation to chicken farmers. This study aimed to compare the general immunity of chicken farmers with those of control subjects with non-agricultural jobs. Blood taken from the farmers and controls was subjected to plasma IgE and IgG subclass measurements. Isolated peripheral blood mononuclear cells (PBMC) were stimulated and cytokine production was measured. Indoor total and respirable dust levels and their endotoxin (LPS) and aflatoxin (AF) levels in the farms were measured. In total, 29 chicken farmers on 19 farms and 14 age- and sex-matched office workers participated. Hematological differences were not observed. The farmers tended to have higher serum IgE and IgG subclass levels with significance for IgG1. The cytokines released by PBMC from farmers indicated skewing toward Type-2 helper T-cell responses: interferon (IFN)-γ:interleukin (IL)-4 and IFNγ:IL-13 ratios were significantly lower than for control PBMC. The farms had 707.1 EU/m3 LPS in total dust, and 15.8 EU/m3 LPS in respirable dust. Farmers exhibited immune skewing towards allergic immune responses that correlated with the LPS levels on their farms. Chicken farmers may be at risk of respiratory allergies due to occupational endotoxin exposure.
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