Yonsei Med J.  2012 May;53(3):593-602. 10.3349/ymj.2012.53.3.593.

Endotoxin Is Not Essential for the Development of Cockroach Induced Allergic Airway Inflammation

• Shin YS
• Sohn JH
• Kim JY
• Lee JH
• Cho SH
• Hong SJ
• Lee JS
• Hong CS
• Park JW

• KMID: 1776996
• DOI: http://doi.org/10.3349/ymj.2012.53.3.593

Abstract

PURPOSE
Cockroach (CR) is an important inhalant allergen and can induce allergic asthma. However, the mechanism by which CR induces airway allergic inflammation and the role of endotoxin in CR extract are not clearly understood in regards to the development of airway inflammation. In this study, we evaluated whether endotoxin is essential to the development of CR induced airway allergic inflammation in mice.
MATERIALS AND METHODS
Airway allergic inflammation was induced by intranasal administration of either CR extract, CR with additional endotoxin, or endotoxin depleted CR extract, respectively, in BALB/c wild type mice. CR induced inflammation was also evaluated with toll like receptor-4 (TLR-4) mutant (C3H/HeJ) and wild type (C3H/HeN) mice.
RESULTS
Intranasal administration of CR extracts significantly induced airway hyperresponsiveness (AHR), eosinophilic and neutrophilic airway inflammation, as well as goblet cell hyperplasia in a dose-dependent manner. The addition of endotoxin along with CR allergen attenuated eosinophilic inflammation, interleukin (IL)-13 level, and goblet cell hyperplasia of respiratory epithelium; however, it did not affect the development of AHR. Endotoxin depletion in CR extract did not attenuate eosinophilic inflammation and lymphocytosis in BAL fluid, AHR and IL-13 expression in the lungs compared to CR alone. The attenuation of AHR, eosinophilic inflammation, and goblet cell hyperplasia induced by CR extract alone was not different between TLR-4 mutant and the wild type mice. In addition, heat inactivated CR extract administration induced attenuated AHR and eosinophilic inflammation.
CONCLUSION
Endotoxin in CR extracts may not be essential to the development of airway inflammation.

Keyword

Cockroach; endotoxin; toll like receptor-4

MeSH Terms

Allergens/*immunology
Animals
Asthma/*chemically induced/*immunology/metabolism
Cockroaches/*immunology
Endotoxins/*immunology
Enzyme-Linked Immunosorbent Assay
Female
Inflammation/*chemically induced/*immunology/metabolism
Interferon-gamma/metabolism
Interleukin-13/metabolism
Interleukin-5/metabolism
Mice
Mice, Inbred BALB C
Respiratory Hypersensitivity/chemically induced/*immunology

Figure

• 

  Fig. 1 Dose-dependency of CR extract-induced asthma model in BALB/c mice. Developments of AHR (A), airway inflammation (B), and IL-13 level in the lungs (C) showed the dose-dependency of CR extracts. (A) CR 75 and 300 µg/mL administrated mice demonstrated significant increases in AHR at MCh concentrations of 25 and 50 mg/mL compared to the sham animal. (B) Total cell number, lymphocyte, eosinophil and neutrophil counts demonstrated statistical differences between sham mice and CR extractadministered mice of any concentration, and (C) IL-13 levels demonstrated significant differences at any concentration of CR extract administration compared to sham mice). *p<0.01, †p<0.05. CR, cockroach; AHR, airway hyperresponsiveness; IL, interleukin; MCh, methacholine.

• 

  Fig. 2 Effects of endotoxin in CR extract-induced asthma model in BALB/c mice. Development of AHR (A) and airway inflammation (B). Cytokine levels (C and D) in the lungs of CR extract administrated BALB/c mice according to endotoxin level. (A) All CR administrated mice showed significant AHR increase at MCh concentrations of 25 and 50 mg/mL compared to sham mice. *p<0.01, †p<0.05. CR, cockroach; AHR, airway hyperresponsiveness; MCh, methacholine.

• 

  Fig. 3 PAS and trichrome staining of CR extract-induced asthma model in BALB/c mice. Representative histology of PAS (A-D) and trichrome (F-I) staining in the lung tissues. Sham mice are A and F. CR induced asthma mice are B and G. Endotoxin depleted CR induced asthma mice are C and H. Endotoxin addition CR induced asthma mice are D and I. The results of PAS and trichrome staining were quantified in E and J. *p<0.05. CR, cockroach; PAS, Periodic Acid-Schiff.

• 

  Fig. 4 CR extractinduced asthma model in TLR-4 wild type and mutant mice. Development of AHR (A), airway inflammation in BAL fluid (B), expression of IL-13 (C), and IFN-γ (D) in the lungs of TLR-4 wild type (C3HeN) and TLR-4 mutant (C3HeJ) mice in the CR induced asthma model. CR administrated C3HeN mice showed significant AHR increase at all MCh concentrations compared to sham mice. *p<0.01, †p<0.05. CR, cockroach; TLR-4, receptor-4; AHR, airway hyperresponsiveness; IL, interleukin; IFN-γ, interferon-gamma; MCh, methacholine.

• 

  Fig. 5 PAS and trichrome staining of the CR extract-induced asthma model in TLR-4 wild type and mutant mice. Representative histology of PAS (A-D) and trichrome (F-I) staining in the lung tissues. No treated wild type mice are A and F. CR treated wild type mice are B and G. No treated TLR-4 mutant mice are C and H. CR treated TLR-4 mutant mice are D and I. The results of PAS and trichrome staining were quantified in E and J. *p<0.01, †p<0.05. CR, cockroach; PAS, Periodic Acid-Schiff; TLR-4, receptor-4.

• 

  Fig. 6 Effects of heat inactivation on the CR extract-induced asthma model in BALB/c mice. Development of AHR (A) and airway inflammation (B) in BALB/c mice administered with heat inactivated CR or non-heat-treated CR. Heat inactivated CR administrated mice showed significant AHR decrease at MCh concentrations of 25 and 50 mg/mL compared to non-heat-treated CR administrated mice. *p<0.01, †p<0.05. CR, cockroach; AHR, airway hyperresponsiveness; MCh, methacholine.

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