Allergy Asthma Immunol Res.  2010 Jul;2(3):199-205. 10.4168/aair.2010.2.3.199.

The Effects of Lactobacillus rhamnosus on the Prevention of Asthma in a Murine Model

• Yu J
• Jang SO
• Kim BJ
• Song YH
• Kwon JW
• Kang MJ
• Choi WA
• Jung HD
• Hong SJ

• KMID: 2133809
• DOI: http://doi.org/10.4168/aair.2010.2.3.199

Abstract

PURPOSE
Lactobacilli are probiotic bacteria that are effective in the management of allergic diseases or gastroenteritis. It is hypothesized that such probiotics have immunoregulatory properties and promote mucosal tolerance. Our goal was to investigate whether Lactobacillus casei rhamnosus Lcr35 could inhibit airway inflammation in an ovalbumin (OVA)-induced murine model of asthma.
METHODS
BALB/c mice aged 6 weeks were used in the present study. Lactobacillus casei rhamnosus Lcr35 was administered daily, starting 1 week prior to the first OVA sensitization (group 1) and 2 days before the first 1% OVA airway challenge (group 2). Mice that received only saline at both sensitization and airway challenge time points were used as negative controls (group 3), and those that had OVA-induced asthma were used as positive controls (group 4). Airway responsiveness to methacholine was assessed, and bronchoalveolar lavage (BAL) was performed. At the endpoint of the study, total IgE as well as OVA-specific IgE, IgG1 and IgG2a in serum was measured by enzyme-linked immunosorbent assay. Lung pathology was also evaluated.
RESULTS
Airway hyperresponsiveness, total cell counts and the proportion of eosinophils in BAL fluid were significantly decreased in group 1 compared with group 4 (P<0.05). Total serum IgE levels were also significantly decreased in group 1 compared with group 4. Serum levels of OVA-specific IgE, IgG1 and IgG(2a) were not significantly influenced by treatment with Lcr35. There was significantly less peribronchial and perivascular infiltration of inflammatory cells in group 1 compared with group 4; however, there were no significant differences in methacholine challenge, BAL, serology or histology between groups 2 and 4.
CONCLUSIONS
Oral treatment with Lcr35 prior to sensitization can attenuate airway inflammation and hyperreactivity in a mouse model of allergic airway inflammation. These results suggest that Lcr35 may have potential for preventing asthma.

Keyword

Asthma; Lactobacillus rhamnosus; probiotics; primary prevention; animal disease model

MeSH Terms

Aged
Animals
Asthma
Bacteria
Bronchoalveolar Lavage
Cell Count
Disease Models, Animal
Enzyme-Linked Immunosorbent Assay
Eosinophils
Gastroenteritis
Humans
Immunoglobulin E
Immunoglobulin G
Inflammation
Lactobacillus
Lactobacillus casei
Lactobacillus rhamnosus
Lung
Methacholine Chloride
Mice
Ovalbumin
Ovum
Primary Prevention
Probiotics
Immunoglobulin E
Immunoglobulin G
Methacholine Chloride
Ovalbumin

Figure

• 

  Fig. 1 Experimental protocol for the induction of allergic asthma and treatment scheme. Four groups of female BALB/c mice were analyzed. Lactobacillus rhamnosus (Lcr35)-treated groups (group 1 and group 2) were sensitized to OVA with adjuvant via intraperitoneal injections, while the control groups (group 3 and group 4) were sensitized with equivalent amounts of saline and OVA, respectively. Group 1: Lactobacillus rhamnosus (Lcr35)-treated group starting 7-day before first OVA sensitization, Group 2: Lactobacillus rhamnosus (Lcr35)-treated group starting 2-day before first 1% OVA challenge, Group 3: saline-sensitized and -challenged group, Group 4: OVA-sensitized and -challenged group.

• 

  Fig. 2 The effects of oral administration of Lactobacillus rhamnosus (Lcr35) on airway hyperresponsiveness. Lactobacillus rhamnosus (Lcr35) administration starting 7-day before first OVA sensitization significantly inhibits OVA-induced airway hyperresponsiveness to MeCh challenge compared with that of positive control group. Data are expressed as the averages of seven mice per group. Values are expressed as mean±SEM. *P<0.05 and **P<0.01, as compared with positive control group (OVA/OVA).

• 

  Fig. 3 Total cell counts in BAL fluid. Oral administration of Lactobacillus rhamnosus (Lcr35) starting 7-day before first OVA sensitization significantly decreases total cell counts in BAL fluid compared with that of positive control group. *P<0.05 and **P<0.01, as compared with positive control group (OVA/OVA). Data are expressed as the averages of seven mice per group. Values are expressed as mean±SEM.

• 

  Fig. 4 Differential cell count in BAL fluid. 24 hr after the last OVA challenge mice were sacrificed and tracheas cannulated for BAL. Cells from BALF were counted, spun onto glass slides and stained with Diff-Quik before microscopy. *P<0.05 and **P<0.01. Data are expressed as the averages of seven mice per group. Values are expressed as mean±SEM.

• 

  Fig. 5 Levels of total IgE, OVA-specific IgE, OVA-specific IgG1 and OVA-specific IgG2a as determined by ELISA in sera of each group. *P<0.05 and **P<0.01. Data are expressed as the averages of seven mice per group. Values are expressed as mean±SEM.

• 

  Fig. 6 Effects of Lactobacillus rhamnosus (Lcr35) treatment on pulmonary inflammation. Representative lung sections from 2-group of Lactobacillus rhamnosus (Lcr35)-treated mice (A: Lcr35 (day -7), B: Lcr35 (day +12)), negative control mice (C: Saline/Saline) and positive control mice (D: OVA/OVA). The data shown are from one representative experiment. Similar results were obtained in three separate experiments using seven mice per group. H&E stain, original magnification: ×100.

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