anti-porcine tlr2-unlabeled rabbit igg  (Santa Cruz Biotechnology)

Journal: BMC Immunology

Article Title: Immunoregulatory effects triggered by immunobiotic Lactobacillus jensenii TL2937 strain involve efficient phagocytosis in porcine antigen presenting cells

doi: 10.1186/s12865-016-0160-1

Figure Lengend Snippet: Generation of porcine blood monocytes-derived dendritic cells (MoDCs). Porcine immature MoDCs were generated from blood monocytes after the differentiation with GM-CSF and IL-4 for 5 days. Maturation of MoDCs was induced by stimulation with LPS. Cell morphology was evaluated by microscopic analysis ( a ). Expression of CD11R1, MHC-II, TLR2, and TLR4 in blood monocytes and mature MoDCs was determined by fluorescent microscopy ( b ). Photos represent data from three independent experiments using different donors. In experiments cells were used in a concentration of 5.0 × 10 6 cells/well. Scale bar = 50 μm

Article Snippet: After removal of the blocking solution, samples were incubated for 16 h at 4 °C in a humidified chamber with one of the following primary antibodies: anti-porcine CD172a-fluorescein isothiocyanate (FITC) SWC3 IgG1 (Southern Biotech), anti-porcine CD11R1-unlabeled mouse IgG1 (AbD Serotec, Kidlington, Oxford, United Kingdom), anti-porcine major histocompatibility complex class II (MHC-II)-unlabeled mouse IgG2a, anti-porcine TLR2-unlabeled rabbit IgG (Santa Cruz); or anti-porcine TLR4-unlabeled rabbit IgG (Santa Cruz).

Techniques: Derivative Assay, Generated, Expressing, Microscopy, Concentration Assay

Journal: BMC Immunology

Article Title: Immunoregulatory effects triggered by immunobiotic Lactobacillus jensenii TL2937 strain involve efficient phagocytosis in porcine antigen presenting cells

doi: 10.1186/s12865-016-0160-1

Figure Lengend Snippet: Role of TLR2 in phagocytosis of immunobiotic Lactobacillus jensenii TL2937 by mononuclear phagocytes from porcine Peyer’s patches (PPMPs). Adherent PPMPs were treated with the immunobiotic strain L. jensenii TL2937 in the presence (anti-TLR2 group) of absence (control group) of blocking anti-TLR2 antibodies. Adherent PPMPs treated with isotype antibodies were used as controls (isotype group). Bacterial phagocytosis by adherent CD172a + CD11R1 + or CD172a − CD11R1 + PPMPs was evaluated using flow cytometric analysis. In experiments cells were used in a concentration of 5.0 × 10 6 cells/well. Values of mean fluorescence intensity (MFI) are shown for each group. The results represent data from three independent experiments using different donors

Article Snippet: Lactobacilli were added to each well at a concentration of 5 × 10 8 cells/ml, and cells were stimulated for 12 h. In addition, unlabeled anti-human TLR2 rabbit IgG (Santa Cruz, CA) was used in blocking experiments.

Techniques: Control, Blocking Assay, Concentration Assay, Fluorescence

Journal: BMC Immunology

Article Title: Immunoregulatory effects triggered by immunobiotic Lactobacillus jensenii TL2937 strain involve efficient phagocytosis in porcine antigen presenting cells

doi: 10.1186/s12865-016-0160-1

Figure Lengend Snippet: Generation of porcine blood monocytes-derived dendritic cells (MoDCs). Porcine immature MoDCs were generated from blood monocytes after the differentiation with GM-CSF and IL-4 for 5 days. Maturation of MoDCs was induced by stimulation with LPS. Cell morphology was evaluated by microscopic analysis ( a ). Expression of CD11R1, MHC-II, TLR2, and TLR4 in blood monocytes and mature MoDCs was determined by fluorescent microscopy ( b ). Photos represent data from three independent experiments using different donors. In experiments cells were used in a concentration of 5.0 × 10 6 cells/well. Scale bar = 50 μm

Article Snippet: Lactobacilli were added to each well at a concentration of 5 × 10 8 cells/ml, and cells were stimulated for 12 h. In addition, unlabeled anti-human TLR2 rabbit IgG (Santa Cruz, CA) was used in blocking experiments.

Techniques: Derivative Assay, Generated, Expressing, Microscopy, Concentration Assay