computer-assisted elispot image analyzer  (Cellular Technology Ltd)

Journal: International Journal of Molecular Sciences

Article Title: The Phagocytosis of Lacticaseibacillus casei and Its Immunomodulatory Properties on Human Monocyte-Derived Dendritic Cells Depend on the Expression of Lc-p75, a Bacterial Peptidoglycan Hydrolase

doi: 10.3390/ijms23147620

Figure Lengend Snippet: The T-cell-polarizing potential of moDCs is modulated by Lc-p75 inactivation in L. casei BL23. Five-day moDCs activated with wild-type (WT) and ∆Lc-p75 mutant L. casei BL23 were washed and then cocultured with autologous T cells from freshly isolated PBLs at a ratio of 1:20 for 3 days. The frequency of cytokine-producing T lymphocytes was measured using IFN-γ ( A ) or IL-17A ( B ) ELISPOT assays. The spot number was counted, or the area covered by the spots was calculated using a computer-assisted ELISPOT image analyzer. The mean value of spot numbers and well areas covered were calculated from 5 independent experiments with 4–6 parallel wells ±SD. One-way ANOVA followed by Tukey’s multiple-comparison test was used for statistical analysis. Significance was defined as * p < 0.05, ** p < 0.01 and **** p < 0.0001 compared to control, non-treated cells (IDC). The differences between WT and ΔLc-p75 were statistically significant and defined as # p < 0.05, ## p < 0.01, and #### p < 0.0001.

Article Snippet: The color development was stopped by tap water, and air-dried plates were analyzed with a computer-assisted ELISPOT image analyzer (Series 1 ImmunoSpot Analyzer, ImmunoSpot Version 4.0 Software Academic, Cellular Technology Limited, Shaker Heights, OH, USA).

Techniques: Mutagenesis, Isolation, Enzyme-linked Immunospot, Comparison