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. 2007 Jan 14;13(2):236-43.
doi: 10.3748/wjg.v13.i2.236.

Correlation between in vitro and in vivo immunomodulatory properties of lactic acid bacteria

Benoit Foligne  1 , Sophie Nutten, Corinne Grangette, Véronique Dennin, Denise Goudercourt, Sabine Poiret, Joelle Dewulf, Dominique Brassart, Annick Mercenier, Bruno Pot
Affiliations

Correlation between in vitro and in vivo immunomodulatory properties of lactic acid bacteria

Benoit Foligne et al. World J Gastroenterol. .

Abstract

Aim: To investigate the correlation between in vitro and in vivo immunomodulation potential of the probiotic strain and its ability to prevent experimental colitis in mice.

Methods: In vitro immunomodulation was assessed by measuring interleukin (IL)-12p70, IL-10, tumor necrosis factor alpha (TNFalpha) and interferon gamma (IFNgamma) release by human peripheral blood mononuclear cells (PBMCs) after 24 h stimulation with 13 live bacterial strains. A murine model of acute TNBS-colitis was next used to evaluate the prophylactic protective capacity of the same set of strains.

Results: A strain-specific in vivo protection was observed. The strains displaying an in vitro potential to induce higher levels of the anti-inflammatory cytokine IL-10 and lower levels of the inflammatory cytokine IL-12, offered the best protection in the in vivo colitis model. In contrast, strains leading to a low IL-10/IL-12 cytokine ratio could not significantly attenuate colitis symptoms.

Conclusion: These results show that we could predict the in vivo protective capacity of the studied lactic acid bacteria (LAB) based on the cytokine profile we established in vitro. The PBMC-based assay we used may thus serve as a useful primary indicator to narrow down the number of candidate strains to be tested in murine models for their anti-inflammatory potential.

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Figures

Figure 1
Figure 1
Strain-specific patterns of IL-10 (A) and IL-12p70 (B) release for various bacterial strains and IL-10/IL-12 ratios (C) for 6 to 12 independent healthy donors. Bars represent the mean ± SE values in pg/mL for 6 to 12 independent healthy donors. Ranked box and whisker plots show the median values and first to third quartiles in boxes. Different letters indicate significant differences according to Mann-Whitney U test (P < 0.05).
Figure 2
Figure 2
IL-10 (A) and IL-12p70 (B) release in 4 distinct individual human PBMC donors and the expression level of IL-10 (C) and IL-12p70 (D). Individual values are represented in pg/mL while IL-10 and IL-12p70 levels are expressed as % of the highest inducer strain. aP < 0.05.
Figure 3
Figure 3
Protective effect of LAB strains against TNBS-induced colitis in BALB/c mice. Results are expressed as a % reduction of the mean macroscopic inflammation of mice treated with LAB as compared to the mean score of non-treated mice. Colitis index was assessed 48 h after TNBS administration. Each bar represents an independent experiment and corresponds to the ratio of control and LAB-treated mice groups (n = 10). aP < 0.05, bP < 0.01 vs TNBS-control group. The horizontal dashed lines indicate the 30% threshold of the uncertain statistical significance.
Figure 4
Figure 4
Hematoxylin/eosin staining of representative cross-sections of murine (BALB/c) distal colon (HE, x 20). A: Normal appearance of the colon from a negative control mice; B: Thickening of the colon wall accompanied with massive inflammatory infiltrate and muscular necrosis 2 d after TNBS-induction of colitis; C: Reduction of histological damage in L. salivarius Ls33-treated mice after TNBS administration; D: Lack of histological improvement of colitis in L. acidophilus NCFM-treated mice after TNBS administration.
Figure 5
Figure 5
Correlation between the average protection against experimental colitis and the mean IL-10/IL-12 ratios obtained in vitro, for all investigated strains. Spearman coefficient (rs = 0.825, P < 0.001). The horizontal dashed lines indicate the 30% threshold of unprotectiveness.
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