Allergy Asthma Immunol Res.  2010 Jan;2(1):34-40. 10.4168/aair.2010.2.1.34.

Preventive effects of mycobacteria and their culture supernatants against asthma development in BALB/c mice

• Han ER
• Choi IS
• Eom SH
• Kim HJ

• KMID: 2260417
• DOI: http://doi.org/10.4168/aair.2010.2.1.34

Abstract

PURPOSE
Live Mycobacterium bovis Bacille Calmette-Guerin (BCG) has a suppressive effect on asthma, but its use in clinical practice may be limited due to adverse reactions. To develop a product that is effective for suppressing asthma with minimal adverse reactions, we investigated whether the heat-killed body or culture supernatants of mycobacteria could also prevent asthma development.
METHODS
Female BALB/c mice were treated with live BCG, the heat-killed body, or culture supernatants of BCG or Mycobacterium tuberculosis intraperitoneally, while sensitizing and provoking with ovalbumin. Then they underwent a methacholine bronchoprovocation test, and the peribronchial inflammatory cell numbers and cytokine levels in splenocyte culture supernatants were assessed.
RESULTS
The airway sensitivity to methacholine decreased significantly after treatment with not only live BCG (30.8 versus 10.0 mg/mL, P<0.001) but also with the culture supernatant (BCG, 23.0 mg/mL, P<0.05; M. tuberculosis, 20.5 mg/mL, P<0.05). In contrast, heat-killed mycobacteria did not effectively decrease airway sensitivity. The peribronchial eosinophil counts and the goblet cell proportions in total epithelial cells decreased significantly in most of the groups. The interferon-gamma/interleukin-5 ratios increased significantly in most of the treatment groups except for the heat-killed groups, and were significantly related to airway sensitivity (r=0.312, P<0.01) and peribronchial eosinophil counts (r=-0.416, P<0.001). Interleukin-17A level was inversely related to airway sensitivity (r=-0.212, P<0.05) and was significantly lower in the live BCG group than in the control (137+/-20 versus 308+/-57 pg/mL, P<0.05).
CONCLUSIONS
BCG and mycobacteria culture supernatants may effectively prevent the development of asthma associated with altered Th1/Th2 cytokines and interleukin-17A levels.

Keyword

Asthma; BCG vaccine; interferons; interleukins; mycobacterium

MeSH Terms

Animals
Asthma
BCG Vaccine
Cell Count
Cytokines
Eosinophils
Epithelial Cells
Female
Goblet Cells
Humans
Interferons
Interleukin-17
Interleukins
Methacholine Chloride
Mice
Mycobacterium
Mycobacterium bovis
Mycobacterium tuberculosis
Ovalbumin
Tuberculosis
BCG Vaccine
Cytokines
Interferons
Interleukin-17
Interleukins
Methacholine Chloride
Ovalbumin

Figure

• 

  Fig. 1 The time course of the experiment. OVA, ovalbumin; IP, intraperitoneal; M. tuberculosis, Mycobacterium tuberculosis; BCG, bacille Calmette-Guérin; HK, heat-killed; Cul-S, culture-supernatant.

• 

  Fig. 2 Dose response curves for methacholine (MCh) challenges. n=10 per group. Penh, enhanced pause; BCG, bacille Calmette-Guérin; TB, Mycobacterium tuberculosis; Cul-S, culture-supernatant. *P<0.05 and **P<0.01 for low dose treatments, and †P<0.05 and ††P<0.01 for high dose treatments compared with the asthma control.

• 

  Fig. 3 Comparisons of the airway responsiveness to methacholine (PC200) between the asthma control group and the other groups. n=10 per group. TB, Mycobacterium tuberculosis; BCG, bacille Calmette-Guérin; HK, heat-killed; Cul-S, culture-supernatant. *P<0.05, **P<0.01, and ***P<0.001 compared with the asthma control.

• 

  Fig. 4 Comparisons of the peribronchial eosinophil numbers and goblet cell hyperplasia in the bronchial epithelium. n=10 per group. TB, Mycobacterium tuberculosis; BCG, bacille Calmette-Guérin; HK, heat-killed; Cul-S, culture-supernatant. *P<0.05, **P<0.01, and ***P<0.001 compared with the asthma control.

• 

  Fig. 5 Comparisons of the interferon (IFN)-γ/interleukin (IL)-5 ratios in the supernatants of concanavalin A-stimulated splenocytes. n=10 per group. TB, Mycobacterium tuberculosis; BCG, bacille Calmette-Guérin; HK, heat-killed; Cul-S, culture-supernatant. *P<0.05, **P<0.01, and ***P<0.05 compared with the asthma control.

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