Interestingly, Lactobacillus-activated MDCs also induced proliferation of autologous CD4+ and CD8+ T cells and induced their secretion of IFN-γ. Similar to our current work, the previous studies found that human MDCs exposed to lactobacilli increased MHC, costimulatory, adhesion, and activation molecules (21–23). You found an error in BacDive? This question is for testing whether or not you are a human visitor and to prevent automated spam submissions. RT-PCR. (B) Treated or untreated MDCs with Lactobacillus species or E. coli LPS were stained with Ab to HLA-DR (green), a fluorescent marker of actin polymer (red), and Hoechst DNA stain (blue). Professional antigen-presenting dendritic cells (DCs) are critical in regulating T cell immune responses at both systemic and mucosal sites. In this study, we examined DC responses to three Lactobacillus species and investigated whether or not these bacteria could induce T cell immune responses in immature human DCs. DCs affect the adaptive immune response by priming T cells to proliferate, become activated, and produce cytokines (12). Likely, this represents one piece of a larger, complex environment in which the responses of multiple immune cells, in combination with the responses of other critical intestinal cell types, such as epithelial cells, are responsible for the overall response to nonpathogenic commensal bacteria (16). Adherent monocytes were cultured with human granulocyte/macrophage colony-stimulating factor (100 ng/ml) and human IL-4 (10 ng/ml) in complete medium consisted of RPMI 1640, 10% FCS, 1% l-glutamine, 1% penicillin/streptomycin, 50 μM 2-mercaptoethanol, 1% sodium pyruvate, and 1% nonessential amino acids (all from GIBCO) for 6 d (17, 18). Int J Syst Evol Microbiol 2000; 50:3-4. MDCs were treated with live Lactobacillus species, E. coli LPS (100 ng/ml), or no supplement for 72 h at 37°C. ). Image credit: Kerry Key (Columbia University, New York, NY). (E–H) MDCs supernatants were harvested and analyzed for proinflammatory cytokines by ELISA. T Cell Proliferation. Sorted naive CD4+ and CD8+ T cells (50 × 103 cells per well in a 96-well plate) were cocultured for 4 d with allogeneic or autologous MDCs, which were treated with live Lactobacillus species or with E. coli LPS. ). Lactobacillus Beijerinck 1901 (Approved Lists 1980). Subsequently, MDCs were cocultured at graded doses with autologous or allogeneic CD4+ or CD8+ T cells (105 cells per well in a 96-well plate) for 4 d in complete RPMI medium 1640 where heat-inactivated 10% human AB+ serum (Gemini Bio-Products, Woodland, CA) replaced FCS. ), as well as the antiinflammatory cytokine IL-10 (Fig. Lactobacillus species were inoculated at 1% and propagated in de Man, Rogosa, and Sharpe broth (MRS, Difco) at 37°C for 15 h. Subsequently, 10 ml of each culture was then transferred to 500 ml of fresh MRS and incubated at 37°C for 8 h until mid-log phase. MDCs were treated with live Lactobacillus species, E. coli LPS (100 ng/ml), or no supplement for 72 h at 37°C. The commensal microbial flora of the intestinal tract harbors Gram-positive and Gram-negative bacteria that may be involved in homeostasis of gut-associated immunity (3–5, 16). These results may indicate that Lactobacillus-activated MDCs induce the activation of effector memory T cells specific for bacterial components. However, we found that the three Lactobacillus species induced IL-12p70, but not IL-10, in MDCs. Validation list no. Lactobacilli activate human dendritic cells that skew T cells toward T helper 1 polarization. Up-Regulation of TLR-2 in MDCs by Lactobacilli. ↵ IL-1β was minimally induced in MDCs treated with L. johnsonii, L. gasseri, or E. coli LPS but was induced in MDCs treated with L. reuteri (Fig. Cells were washed and analyzed by flow cytometry. adj. Flow Cytometry. NOTE: We only request your email address so that the person you are recommending the page to knows that you wanted them to see it, and that it is not junk mail. Experiments were performed at least three times with similar results. We do not capture any email address. FACS and confocal analysis of MDCs treated with different Lactobacillus species. Moreover, IL-6 was induced in MDCs at similar levels when treated with each Lactobacillus species or E. coli LPS (Fig. In contrast, MDCs treated with L. reuteri or L. johnsonii induced IL-10 in allogeneic CD4+ T cells. Isolated CD4+ T cells were negatively purified by the depletion of CD8+, CD19+, CD56+, CD1a+, and CD14+ cells by using specific Ab conjugated beads (Miltenyi Biotec, Auburn, CA). All experiments were repeated at least three times. In this way, the gut may maintain a balance between inflammatory responses to pathogens and natural intestinal homeostasis and function. Lethally-irradiated Lactobacillus species (10 μg/ml) also activated MDCs to secrete IL-12 p70 and IL-18, but not IL-10, respectively (data not shown). Image credit: Noah Whiteman (University of California, Berkeley, CA). Cells were pulsed with 0.5 μCi of [3H]thymidine per well (New England Nuclear). These data suggest that lactobacilli may deliver signals in MDCs through TLR-2, thereby promoting the activation of these cells. Experiments were repeated at least three times with similar results. All three Lactobacillus species modulated the MDC phenotype by up-regulating HLA-DR, activation of costimulatory molecules CD40, CD80, CD83, and CD86, and down-regulation of CD1a (Fig. Here we demonstrate an alternative approach to treat contamination using a novel peptidoglycan hydrolase (LysKB317) isolated from a bacteriophage, EcoSau. ) were examined. Species designations and categories of the 473 non-enterococcal LAB isolates included in this study are listed in Table 1. ) and induced the secretion of IFN-γ and IL-2, but not IL-4 or IL-13 by naive CD4+CD62L+CD45R A+CD45RO– or CD8+CD62L+CD45RA+CD45RO– T cells (Fig. Back W, Bohak I, Ehrmann M, Ludwig W, Pot B, Kersters K, Schleifer KH. AB289249 Assigned by: However, for autologous naive CD4+ or CD8+ T cells, Lactobacillus-activated MDCs did not induce their proliferation or activation. Linking: The mechanisms underlying the different MDC responses induced by varying Lactobacillus species and strains are unknown.

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