Journal: Bioscience of Microbiota, Food and Health

Article Title: Identification of Immunopotentiating Lactic Acid Bacteria that Induce Antibody Production by in vitro Stimulated Human Peripheral Blood Mononuclear Cells

doi: 10.12938/bmfh.31.7

Figure Lengend Snippet: The effects of LAB on total antibody production by human PBMCs stimulated in vitro . LLME-treated PBMCs were stimulated in vitro with β-lactoglobulin in the presence of IL-2, IL-4 and LAB or CpG2006 ODN. After 7 days of culture, the amount of IgM, IgG, and IgA antibodies produced were evaluated by ELISA.

Article Snippet: LLME-treated PBMCs (5 × 10 6 cells) were sensitized with β-lactoglobulin (10 μg/mL; Wako, Osaka, Japan) in the presence of IL-2 (10 units/mL; R&D Systems, Minneapolis, MN, USA), IL-4 (10 ng/mL; PeproTech, London, UK) and CpG2006 oligodeoxynucleotide (CpG2006 ODN, 52-TCGTCGTTTTGTCGTTTTGTCGT-32, 1 μg/mL; Sigma-Genosys, Ishikari, Japan) or LAB (10 μg/mL) and cultured in eRDF medium supplemented with 10% heat-inactivated FBS and 2-ME (50 μM) for 7 days.

Techniques: In Vitro, Produced, Enzyme-linked Immunosorbent Assay

Journal: Bioscience of Microbiota, Food and Health

Article Title: Identification of Immunopotentiating Lactic Acid Bacteria that Induce Antibody Production by in vitro Stimulated Human Peripheral Blood Mononuclear Cells

doi: 10.12938/bmfh.31.7

Figure Lengend Snippet: The effects of LAB on antigen-specific antibody production by human PBMCs stimulated in vitro . LLME-treated PBMCs were stimulated in vitro with β-lactoglobulin in the presence of IL-2, IL-4, and LAB or CpG2006 ODN. After 7 days of culture, B cells producing antigen-specific antibody were detected by an ELISPOT assay. Wells were coated with β-lactoglobulin for detecting antigen-specific antibody-producing cells or with fish gelatin (FG) for non-specific antibody-producing cells. The number of spots in A were counted using the ImageJ software and depicted in B. NC, negative control.

Article Snippet: LLME-treated PBMCs (5 × 10 6 cells) were sensitized with β-lactoglobulin (10 μg/mL; Wako, Osaka, Japan) in the presence of IL-2 (10 units/mL; R&D Systems, Minneapolis, MN, USA), IL-4 (10 ng/mL; PeproTech, London, UK) and CpG2006 oligodeoxynucleotide (CpG2006 ODN, 52-TCGTCGTTTTGTCGTTTTGTCGT-32, 1 μg/mL; Sigma-Genosys, Ishikari, Japan) or LAB (10 μg/mL) and cultured in eRDF medium supplemented with 10% heat-inactivated FBS and 2-ME (50 μM) for 7 days.

Techniques: In Vitro, Enzyme-linked Immunospot, Software, Negative Control

Journal: Bioscience of Microbiota, Food and Health

Article Title: Identification of Immunopotentiating Lactic Acid Bacteria that Induce Antibody Production by in vitro Stimulated Human Peripheral Blood Mononuclear Cells

doi: 10.12938/bmfh.31.7

Figure Lengend Snippet: Activation profile of PBMCs stimulated in vitro in the presence of CpG2006 ODN or L32. A: After LLME-treated PBMCs were stimulated in vitro as described above, CD3 + T cells were isolated, and CD69 expression in CD4 + /CD4– T cells was monitored by a flow cytometer. B: After LLME-treated PBMC were stimulated in vitro as described above, CD19 + B cells were isolated, and CD80 and CD86 expression was monitored by a flow cytometer.

Article Snippet: LLME-treated PBMCs (5 × 10 6 cells) were sensitized with β-lactoglobulin (10 μg/mL; Wako, Osaka, Japan) in the presence of IL-2 (10 units/mL; R&D Systems, Minneapolis, MN, USA), IL-4 (10 ng/mL; PeproTech, London, UK) and CpG2006 oligodeoxynucleotide (CpG2006 ODN, 52-TCGTCGTTTTGTCGTTTTGTCGT-32, 1 μg/mL; Sigma-Genosys, Ishikari, Japan) or LAB (10 μg/mL) and cultured in eRDF medium supplemented with 10% heat-inactivated FBS and 2-ME (50 μM) for 7 days.

Techniques: Activation Assay, In Vitro, Isolation, Expressing, Flow Cytometry