cfse controls  (Bio-Rad)

Journal: Cell Death & Disease

Article Title: PP2A-dependent control of transcriptionally active FOXO3a in CD8 + central memory lymphocyte survival requires p47 phox

doi: 10.1038/cddis.2012.118

Figure Lengend Snippet: Significantly fewer CD4 + and CD8 + lymphocytes accumulate in the infected spleens of p47 phox−/− mice during primary L. monocytogenes infection. WT ( ) and p47 phox−/− ( ) mice were infected with 5 × 10 4 CFU (0.1 LD 50 ) rLM-OVA. On the indicated post infection day the mice were euthanized and single-cell splenocyte cultures were incubated for 3–5 h with 1 μ M OVA 323-339 and/or OVA 257-264 peptides for CD4 and CD8 T lymphocyte stimulation, respectively. Monensin was added for the final 2 h of culture, and the cells were harvested and stained for surface CD4, CD8, and intracellular IFN γ expression. ( a ) Percentage of CD4 + and CD8 + lymphocytes, and the percentage of CD4 + and CD8 + lymphocytes expressing IFN γ . The data are the mean (±S.E.M.) for six individual mice from two separate experiments with three of each genotype/experiment. ( b ) p47 phox−/− CD8 + T lymphocytes effectively kill target cells. On post infection days 3 and 7, as indicated, spleen-derived CD8 + T cells were co-cultured with CFSE-labeled EL4 or E.G7-OVA target cells at the indicated effect: target cell ratios for 4 h. The results are presented as percentage of PI + E.G7-OVA or EL4 target cells. Results are the mean (±S.E.M.) of three independent experiments including three of each genotype/experiment. ( c ) On post infection day 3, the percentage of non-viable EMA + CD4 + or EMA + CD8 + lymphocytes was determined. The data are the mean (±S.E.M.) percentage for five individual mice. * P <0.005, ** P <0.05

Article Snippet: The killing assay was performed by co-culturing CD8 + lymphocytes with CFSE-labeled control EL4 cells (ATCC, Manassas, VA, USA) or the OVA-expressing EL4 derivative cell line E.G7-OVA (ATCC) at the indicated effect: target cells ratios.

Techniques: Infection, Incubation, Staining, Expressing, Derivative Assay, Cell Culture, Labeling

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Functional endogenous cytotoxic T lymphocytes are generated to multiple antigens co-expressed by progressing tumors; after intra-tumoral IL-2 therapy these effector cells eradicate established tumors

doi: 10.1007/s00262-005-0086-6

Figure Lengend Snippet: Tumor antigen is presented in tumor-draining lymph nodes. CFSE-labeled, SIINFEKL-specific T cells from OT-1 mice were adoptively transferred at days 5 or 6 into mice inoculated s.c on day 0 with Lewis lung, LLsOVA or LLpoly tumor cells. a CFSE+CD8+ cells were reisolated from the DLN 3 days post-transfer for analysis. Representative FACs profiles from individual mice are shown (n=6 mice/time point) b The mean (±SE) percentage of CFSE+CD8+ cells proliferating in the DLN is indicated on the histogram: there was no statistical difference between LLsOVA and LLpoly tumor-bearing mice

Article Snippet: Analysis of 2000 (or more) CFSE-labeled control targets or CD8 + cells was performed on a FACScan (Becton Dickinson, Mountain View, CA, USA) using Cell Quest software.

Techniques: Labeling