A novel mouse model of diisocyanate-induced asthma showing allergic-type inflammation in the lung after inhaled antigen challenge

doi:10.1067/mai.2002.123533

Journal of Allergy and Clinical Immunology

Volume 109, Issue 5, May 2002, Pages 873-878

https://doi.org/10.1067/mai.2002.123533 Get rights and content

Abstract

Background: Exposure to diisocyanates, a group of highly reactive, low-molecular-weight compounds, is a major cause of occupational asthma. In contrast to mouse models of atopic asthma, previous mouse models of diisocyanate-induced asthma have failed to show lung inflammation with characteristics of human disease. Objective: Our goal was to establish a novel mouse model of diisocyanate-induced asthma in which lung inflammation reminiscent of that seen in human asthma is generated after inhaled antigen challenge. Methods: BALB/c mice were epicutaneously sensitized to hexamethylene diisocyanate (HDI) and then challenged with an HDI-protein conjugate administered by means of an intranasal droplet. Results: HDI sensitization resulted in development of contact hypersensitivity and HDI-specific antibody production. Most importantly, however, vigorous inflammatory responses with characteristics of human asthma were generated in the lung after inhaled HDI challenge. Challenge of sensitized, but not unsensitized, mice resulted in airway eosinophilia, mucus hypersecretion, and production of T_H1-type (IFN-γ) and T_H2-type (IL-4, IL-5, and IL-13) cytokines by lung inflammatory cells. Despite the mixed T_H1/T_H2 response induced by HDI sensitization, use of cytokine-deficient mice revealed that airway eosinophilia was mediated by T_H2 cytokines and not by IFN-γ. Conclusion: We report a novel mouse model of diisocyanate-induced asthma that, in contrast to previous models, demonstrates antigen-induced lung inflammation with characteristics of human disease. This model will allow investigation of the immunopathogenesis of diisocyanate-induced asthma and should provide insight into this common form of occupational disease. (J Allergy Clin Immunol 2002;109:873–8.)

Section snippets

Animals and epicutaneous sensitization

BALB/c mice were purchased from The National Cancer Institute (Frederick, Md). BALB/c-Il4^tm2Nnt (IL-4-/-) mice were purchased from The Jackson Laboratory (Bar Harbor, Me). BALB/c IL-13-/- mice were provided by A. McKenzie. IFN-γ receptor-deficient (IFN-γR-/-) mice were provided by J. Aguet and backcrossed onto BALB/c. Female mice 9 to 12 weeks of age were used in all experiments. Backs of mice were shaved with electric clippers 1 to 2 days before exposure to HDI (0.01%–10%) diluted in a 4:1

CHS and antibody responses in HDI-sensitized mice

As shown in Fig 1, A , mice exposed to either 0.1% or 1.0% HDI on days 0 and 7 had significant ear-swelling responses after challenge on day 12.

. CHS, serum antibody, and airway inflammatory responses in HDI-sensitized mice. A, Ears of sensitized mice were challenged with HDI, and peak swelling at 24 hours is shown. Data are reported as means ± SEM of 4 to 5 mice per group (representative experiment, 1 of 3). B and C, HDI-sensitized or unsensitized mice were challenged with inhaled HDI-MSA.

Discussion

In the present study we describe a novel mouse model of diisocyanate-induced asthma. To our knowledge, this is the first such model to demonstrate lung inflammation with characteristics of human asthma, including eosinophilia and mucus hypersecretion. The lung inflammation in our model was present specifically in sensitized animals. Scheerens et al7, 8 have reported increased airway reactivity after intratracheal challenge of toluene diisocyanate-sensitized mice. However, only nonspecific,

Acknowledgements

We thank Ms Heather MacLeod for technical assistance and Dr Robert Homer for his input regarding the histologic appearance of lung inflammatory infiltrates.

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Toluene diisocyanate (TDI), a known human asthmagen, was investigated in skin-sensitized Brown Norway rats for its concentration × time (C × t)-response relationship on elicitation-based endpoints. The major goal of study was to determine the elicitation inhalation threshold dose in sensitized, re-challenged Brown Norway rats, including the associated variables affecting the dosimetry of inhaled TDI-vapor in rats and as to how these differences can be translated to humans. Attempts were made to duplicate at least some traits of human asthma by using skin-sensitized rats which were subjected to single or multiple inhalation-escalation challenge exposures. Two types of dose-escalation protocols were used to determine the elicitation-threshold C × t; one used a variable C (C_var) and constant t (t_const), the other a constant C (C_const) and variable t (t_var). The selection of the “minimal irritant" C was based an ancillary pre-studies. Neutrophilic granulocytes (PMNs) in bronchoalveolar lavage fluid (BAL) were considered as the endpoint of choice to integrate the allergic pulmonary inflammation. These were supplemented by physiological measurements characterizing nocturnal asthma-like responses and increased nitric oxide in exhaled breath (eNO). The C_const × t_var regimen yielded the most conclusive dose–response relationship as long C was high enough to overcome the scrubbing capacity of the upper airways. Based on ancillary pre-studies in naïve rats, the related human-equivalent respiratory tract irritant threshold concentration was estimated to be 0.09 ppm. The respective 8-h time-adjusted asthma-related human-equivalent threshold C × t-product (dose), in ‘asthmatic’ rats, was estimated to be 0.003 ppm. Both thresholds are in agreement of the current ACGIH TLV^® of TDI and published human evidence. In summary, the findings from this animal model suggest that TDI-induced respiratory allergy is likely to be contingent on two interlinked, sequentially occurring mechanisms: first, dermal sensitizing encounters high enough to cause systemic sensitization. Second, when followed by inhalation exposure(s) high enough to initiate and amplify an allergic airway inflammation, then a progression into asthma may occur. This bioassay requires an in-depth knowledge on respiratory tract dosimetry and irritation of the involved test substance to clearly understand the dosimetry causing C- and/or C × t-dependent respiratory tract irritation and eventually asthma.
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For example, skin exposure to some occupational chemicals induces strong TH2-skewed responses, whereas others induce TH1 skewed responses.62–64 Exposure dose further influences the outcome of skin exposure, which may be nonlinear and/or paradoxically limited at higher doses.45,47 Increasing recognition of the potential for occupational skin exposure to contribute to OA has spawned the development of animal models to further investigate potential pathogenic mechanisms.

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^☆: Supported by National Institutes of Health grants R01-HL65209 (K. Bottomly and C. Herrick), P50-HL56389 and K24-ES00355 (C. Redlich), and RO1-HL62622 (A. Wisnewski). C. Herrick was supported by a Dermatology Foundation Clinical Career Development Award.

^☆☆: Reprint requests: Christina A. Herrick, MD, PhD, Department of Dermatology, Yale University School of Medicine, 333 Cedar St, PO Box 208059, New Haven, CT 06520-8059.

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Enviornmental and Occupational DisordersA novel mouse model of diisocyanate-induced asthma showing allergic-type inflammation in the lung after inhaled antigen challenge☆,☆☆

Abstract

Section snippets

Animals and epicutaneous sensitization

CHS and antibody responses in HDI-sensitized mice

Discussion

Acknowledgements

Asthma

N Engl J Med

IL-4 and IL-5 mRNA and protein in bronchial biopsies from patients with atopic and nonatopic asthma: evidence against “intrinsic” asthma being a distinct immunopathologic entity

Am J Respir Crit Care Med

Comparison of the immunopathology of extrinsic, intrinsic and occupational asthma

J Investig Allergol Clin Immunol

Occupational asthma

N Engl J Med

Overview of diisocyanate occupational asthma

Toxicology

Occupational asthma as identified by the Surveillance of Work-related and Occupational Respiratory Diseases programme in South Africa

Clin Exp Allergy

Toluene diisocyanate-induced in vitro tracheal hyperreactivity in the mouse

Am J Respir Crit Care Med

Long-term topical exposure to toluene diisocyanate in mice leads to antibody production and in vivo airway hyperresponsiveness three hours after intranasal challenge

Am J Respir Crit Care Med

Importance of inflammatory and immune components in a mouse model of airway reactivity to toluene diisocyanate (TDI)

Clin Exp Allergy

Activated T lymphocytes and eosinophils in the bronchial mucosa in isocyanate-induced asthma

J Allergy Clin Immunol

Mechanisms of occupational asthma

Clin Exp Allergy

Occupational asthma and extrinsic allergic alveolitis due to isocyanates: current status and perspectives

Br J Ind Med

Th2 responses induced by epicutaneous or inhalational protein exposure are differentially dependent on IL-4

J Clin Invest

Induction of respiratory hypersensitivity to diphenylmethane 4,4'-diisocyanate (MDI) in guinea pigs. Influence of the route of exposure

Toxicology

IL-4 promotes airway eosinophilia by suppressing IFN-gamma production: defining a novel role for IFN-gamma in the regulation of allergic airway inflammation

J Immunol

Enviornmental and Occupational Disorders
A novel mouse model of diisocyanate-induced asthma showing allergic-type inflammation in the lung after inhaled antigen challenge☆,☆☆