In vitro immunomodulatory activity of Lactobacillus fermentum CECT5716 and Lactobacillus salivarius CECT5713: Two probiotic strains isolated from human breast milk

Department of Physiology, Faculty of Pharmacy, University of Barcelona, Spain.
Immunobiology (Impact Factor: 3.04). 02/2010; 215(12):996-1004. DOI: 10.1016/j.imbio.2010.01.004
Source: PubMed


Commensal bacteria, including some species of lactobacilli commonly present in human breast milk, appear to colonize the neonatal gut and contribute to protection against infant infections, suggesting that lactobacilli could potentially modulate immunity. In this study, we evaluated the potential of two Lactobacillus strains isolated from human milk to modulate the activation and cytokine profile of peripheral blood mononuclear cell (PBMC) subsets in vitro. Moreover, these effects were compared to the same probiotic species of non-milk origin. Lactobacillus salivarius CECT5713 and Lactobacillus fermentum CECT5716 at 10⁵, 10⁶ and 10⁷ bacteria/mL were co-cultured with PBMC (10⁶/mL) from 8 healthy donors for 24 h. Activation status (CD69 and CD25 expressions) of natural killer (NK) cells (CD56+), total T cells (CD3+), cytotoxic T cells (CD8+) and CD4+ T cells was determined by flow cytometry. Regulatory T cells (Treg) were also quantified by intracellular Foxp3 evaluation. Regarding innate immunity, NK cells were activated by addition of both Lactobacillus strains, and in particular, the CD8+ NK subset was preferentially induced to highly express CD69 (~90%, p<0.05). With respect to acquired immunity, approximately 9% of CD8+ T cells became activated after co-cultivation with L. fermentum or L salivarius. Although CD4+ T cells demonstrated a weaker response, there was a preferential activation of Treg cells (CD4+CD25+Foxp3+) after exposure to both milk probiotic bacteria (p<0.05). Both strains significantly induced the production of a number of cytokines and chemokines, including TNFα, IL-1β, IL-8, MIP-1α, MIP-1β, and GM-CSF, but some strain-specific effects were apparent. This work demonstrates that L salivarius CECT5713 and L. fermentum CECT5716 enhanced both natural and acquired immune responses, as evidenced by the activation of NK and T cell subsets and the expansion of Treg cells, as well as the induction of a broad array of cytokines.

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Available from: Honglin Dong, Aug 28, 2014
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    • "Among these, Lactobacillus fermentum CECT 5716 has found commercial applications due to its repertoire of desirable properties, including its ability to inhibit the growth of a wide spectrum of pathogenic bacteria (Martín et al. 2005a; Olivares et al. 2006), its role in supporting maturation of the infant immune system by acting on both innate and acquired immunity through a variety of mechanisms (Díaz-Ropero et al. 2006; Olivares et al. 2007; Pérez-Cano et al. 2010), its anti-inflammatory activity (Mañé et al. 2009), its high rate of survival in conditions simulating those in the gastrointestinal tract (Martín et al. 2005a), prebiotic metabolism (Bañuelos et al. 2008), resistance against diet polyphenols (Cueva et al. 2010), its safety (Lara-Villoslada et al. 2009; Gil-Campos et al. 2012), and its in vivo efficacy to reduce the incidence of gastrointestinal and upper respiratory tract infections in infants (Maldonado et al. 2012). L. fermentum, a heterofermentative species of the genus Lactobacillus, is a normal inhabitant of the human gastrointestinal tract, including that of breast-fed infants (Grover et al. 2013). "
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    ABSTRACT: Lactobacillus fermentum CECT 5716, isolated from human milk, has immunomodulatory, anti-inflammatory, and anti-infectious properties, as revealed by several in vitro and in vivo assays, which suggests a strong potential as a probiotic strain. In this work, some phenotypic properties of L. fermentum CECT 5716 were evaluated, and the genetic basis for the obtained results was searched for in the strain genome. L. fermentum CECT 5716 does not contain plasmids and showed neither bacteriocin nor biogenic amine biosynthesis ability but was able to produce organic acids, glutathione, riboflavin, and folates and to moderately stimulate the maturation of mouse dendritic cells. No prophages could be induced, and the strain was sensitive to all antibiotics proposed by European Food Safety Authority (EFSA) standards, while no transmissible genes potentially involved in antibiotic resistance were detected in its genome. Globally, there was an agreement between the phenotype properties of L. fermentum CECT 5716 and the genetic information contained in its genome.
    Full-text · Article · Feb 2015 · Applied Microbiology and Biotechnology
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    • "Some lactobacilli isolated from human milk have been characterized and shown to have probiotic potential [13–15]. Specifically, L. salivarius CECT5713 that was isolated from human milk and infant feces of a healthy mother-child pair has been shown to have remarkable probiotic potential because it had high rate of survival in simulated gastrointestinal tract conditions and strong adherence to mucus and intestinal cells in vitro, stimulated the expression of mucin-encoding genes, and produced antimicrobial compounds [14–17]. More recently, its complete genome has been sequenced [18], and its genetic features, such as proteins potentially involved in human molecular mimetism, may explain its immunomodulatory, anti-inflammatory, and anti-infectious properties [19, 20]. "
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    ABSTRACT: Cheeses have been proposed as a good alternative to other fermented milk products for the delivery of probiotic bacteria to the consumer. The objective of this study was to assess the survival of two Lactobacillus salivarius strains (CECT5713 and PS2) isolated from human milk during production and storage of fresh cheese for 28 days at 4°C. The effect of such strains on the volatile compounds profile, texture, and other sensorial properties, including an overall consumer acceptance, was also investigated. Both L. salivarius strains remained viable in the cheeses throughout the storage period and a significant reduction in their viable counts was only observed after 21 days. Globally, the addition of the L. salivarius strains did not change significantly neither the chemical composition of the cheese nor texture parameters after the storage period, although cheeses manufactured with L. salivarius CECT5713 presented significantly higher values of hardness. A total of 59 volatile compounds were identified in the headspace of experimental cheeses, and some L. salivarius-associated differences could be identified. All cheeses presented good results of acceptance after the sensory evaluation. Consequently, our results indicated that fresh cheese can be a good vehicle for the two L. salivarius strains analyzed in this study.
    Full-text · Article · May 2014 · BioMed Research International
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    • "Extensive studies have been performed to determine how lactobacilli and bifidobacteria regulate infant gut immunity [24], [25]. However, few studies have focused on E.faecalis, which is the first colonizer in the human GI tract [6], [7]. "
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    ABSTRACT: Colonizing commensal bacteria after birth are required for the proper development of the gastrointestinal tract. It is believed that bacterial colonization pattern in neonatal gut affects gut barrier function and immune system maturation. Studies on the development of faecal microbiota in infants showed that the neonatal gut was first colonized with enterococci followed by other microbiota such as Bifidobacterium. Other studies showed that babies who developed allergy were less often colonized with Enterococcus during the first month of life as compared to healthy infants. Many studies have been conducted to elucidate how bifidobacteria or lactobacilli, some of which are considered probiotic, regulate infant gut immunity. However, fewer studies have been focused on enterococi. In our study, we demonstrate that E. faecalis, isolated from healthy newborns, suppress inflammatory responses activated in vivo and in vitro. We found E. faecalis attenuates proinflammatory cytokine secretions, especially IL-8, through JNK and p38 signaling pathways. This finding shed light on how the first colonizer, E.faecalis, regulates inflammatory responses in the host.
    Full-text · Article · May 2014 · PLoS ONE
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