h4b4 mouse anti human lamp  (Developmental Studies Hybridoma Bank)

Journal: PLoS ONE

Article Title: Isoflavones, Genistein and Daidzein, Regulate Mucosal Immune Response by Suppressing Dendritic Cell Function

doi: 10.1371/journal.pone.0047979

Figure Lengend Snippet: Flow cytometric gating strategy for DC-NK cell co-culture: Human MDDCs were activated with 100 ng/mL LPS, 1 µg/ml CT, or 100 ng/mL TNF-α +/− 100 µM genistein (G100) or daidzein (D100) for 18 h, washed and incubated with allogenic NK cells for 2 h in culture medium with FITC conjugated anti-human LAMP-1 and LAMP-2 Abs (2.5 µg/ml). The cells were then stained for CD83, CD56, CD69 and live/dead discriminator and analyzed by flow cytometry (A). Effector autologous NK cells were incubated with stimulant +/− isoflavone-treated target DCs (as above) in ratios ranging from 20∶1 to 0.63∶1 as indicated and stained as above. The percentage of LAMP-1/2 hi cells were plotted for each E:T ratio and stimulation condition as line graph (B). Effector allogenic NK cells were incubated with stimulant +/− isoflavones treated target DCs (as above) in 1∶1 ratio and stained as above (C & D). The percentage of LAMP-1/2 hi cells (C) and the percentage of DC cell death (D) are plotted for all the conditions and shown as bar graphs. DMSO - vehicle control for genistein and daidzein. The data pooled from 7 independent experiments from different cell donors for LPS stimulation, 5 experiments for CT stimulations and 2 experiments for TNF stimulations. DMSO : vehicle control. Statistical significance is indicated by * (P≤0.05) or ** (P≤0.01) (paired student t test).

Article Snippet: FITC-conjugated mouse anti-human CD107a (lysosome-associated membrane protein (LAMP)-1) and CD107b (LAMP)-2 antibodies (BD Biosciences) (2.5 µg/ml) were included in culture media.

Techniques: Co-Culture Assay, Incubation, Staining, Flow Cytometry