Allergy Asthma Immunol Res.  2012 May;4(3):150-156. 10.4168/aair.2012.4.3.150.

Asthma Prevention by Lactobacillus Rhamnosus in a Mouse Model is Associated With CD4+CD25+Foxp3+ T Cells

• Jang SO
• Kim HJ
• Kim YJ
• Kang MJ
• Kwon JW
• Seo JH
• Kim HY
• Kim BJ
• Yu J
• Hong SJ

• KMID: 2167067
• DOI: http://doi.org/10.4168/aair.2012.4.3.150

Abstract

PURPOSE
Probiotic bacteria can induce immune regulation or immune tolerance in allergic diseases. The underlying mechanisms have been recently investigated, but are still unclear. The aim of this study was to evaluate the protective effects of the probiotic Lactobacillus rhamnosus (Lcr35) in a mouse model of asthma and to identify its mechanism of action.
METHODS
Lcr35 was administered daily by the oral route at a dosage of 1x10(9) CFU/mouse in BALB/c mice for 7 days before the first sensitization. Clinical parameters and regulatory T (Treg) cells were examined. The role of CD4+CD25+Foxp3+ Treg cells was analyzed using a Treg cell-depleting anti-CD25 monoclonal antibody (mAb).
RESULTS
Airway hyperresponsiveness, total IgE production, pulmonary eosinophilic inflammation, and splenic lymphocyte proliferation were suppressed after Lcr35 treatment. Th1 (IFN-gamma) and Th2 (IL-4, IL-5, and IL-13) cytokines in the serum were suppressed, and the percentage of CD4+CD25+Foxp3+ Treg cells in the spleen was significantly increased in the Lcr35 treatment group. Anti-CD25 mAb administration abolished the protective effects of Lcr35, indicating that CD4+ CD25+Foxp3+ Treg cells are essential in mediating the activity of Lcr35.
CONCLUSIONS
Oral administration of Lcr35 attenuated the features of allergic asthma in a mouse model and induced immune regulation by a CD4+CD25+Foxp3+ Treg cell-mediated mechanism.

Keyword

Asthma; probiotics; mice; T-lymphocytes, Regulatory

MeSH Terms

Administration, Oral
Animals
Asthma
Bacteria
Cytokines
Eosinophils
Immune Tolerance
Immunoglobulin E
Inflammation
Interleukin-5
Lactobacillus
Lactobacillus rhamnosus
Lymphocytes
Mice
Negotiating
Probiotics
Spleen
T-Lymphocytes
T-Lymphocytes, Regulatory
Cytokines
Immunoglobulin E
Interleukin-5

Figure

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  Fig. 1 Effect of oral Lcr35 treatment on allergic asthma in a mouse model. (A) Airway hyperresponsiveness; (B) Eosinophil proportion in bronchoalveolar lavage fluid; (C) Peribronchial and perivascular lung inflammation score; (D) Lung pathology. *P<0.05 and **P<0.01. Negative control: Saline (i.p.)/saline challenge by inhalation. Lcr35-treated: OVA (i.p.)/OVA challenge by inhalation+Lcr35 (oral administration). Positive control: OVA (i.p.)/OVA challenge by inhalation. Lcr35, Lactobacillus rhamnosus; i.p., intraperitoneal; OVA, ovalbumin.

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  Fig. 2 Effect of oral Lcr35 treatment on the suppression of OVA-specific splenic T cell proliferation in a mouse model. *P<0.05 and **P<0.01. Negative control: Saline (i.p.)/saline challenge by inhalation. Lcr35-treated: OVA (i.p.)/OVA challenge by inhalation+Lcr35 (oral administration). Positive control: OVA (i.p.)/OVA challenge by inhalation. Lcr35, Lactobacillus rhamnosus; OVA, ovalbumin; i.p., intraperitoneal.

• 

  Fig. 3 Effect of oral Lcr35 treatment on the CD4+CD25+Foxp3+ Treg population in a mouse model. (A) Flow cytometric analysis of T cells in the mouse spleen. Plots show cells expressing CD4, CD25, and intracellular Foxp3. Numbers inside the histograms indicate the percentage of cells; (B) Relative proportion of CD4+CD25+Foxp3+ Treg cells among total splenic T cells after Lcr35 treatment in comparison with cells obtained from the positive control (Group 3). *P<0.05. Negative control: Saline (i.p.)/saline challenge by inhalation. Lcr35-treated: OVA (i.p.)/OVA challenge by inhalation+Lcr35 (oral administration). Positive control: OVA (i.p.)/OVA challenge by inhalation. Lcr35, Lactobacillus rhamnosus; i.p., intraperitoneal; OVA, ovalbumin.

• 

  Fig. 4 Effect of anti-CD25 mAb treatment on allergic asthma in a mouse model after oral Lcr35-treatment. (A) Airway hyperresponsiveness; (B) Relative proportion of CD4+CD25+Foxp3+ Treg cells among total splenic T cells; (C) Comparison of pulmonary inflammation scores; (D) Lung pathology (H&E stain; ×100). *P<0.05. Lcr35-treated: OVA (i.p.)/OVA challenge by inhalation+Lcr35 (oral administration). Anti-CD25: OVA (i.p.)/OVA challenge by inhalation+Lcr35 (oral administration)+anti-CD25 Ab (i.p.). Negative control: Saline (i.p.)/saline challenge by inhalation. Positive control: OVA (i.p.)/OVA challenge by inhalation. Lcr35, Lactobacillus rhamnosus; OVA, ovalbumin; i.p., intraperitoneal.

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