Journal: Scientific Reports

Article Title: Adoptive T-cell therapy improves treatment of canine non–Hodgkin lymphoma post chemotherapy

doi: 10.1038/srep00249

Figure Lengend Snippet: T cells isolated from PBMC of healthy dogs were numerically expanded on γ-irradiated OKT3-loaded aAPC at a T-cell : aAPC ratio of 2:1. T cells were re-stimulated with aAPC every 7 days. Recombinant human IL-21 and/or rhIL-2 were added to the co-culture of T cells on aAPC as shown. (a) Parental K562 cells were genetically modified to co-express human CD64 and the human T-cell co-stimulatory molecules CD86, CD137L, and membrane-bound IL-15 (co-expressed with EGFP) and cloned (CLN4). OKT3 was loaded via introduced CD64 on CLN4 and detected with a Fab-specific antibody. CLN4 is shown as gray-open histograms and K562 parental cells as black-open histograms. (b) Numeric expansion of T cells on γ-irradiated OKT3-loaded aAPC (added every 7 days) in the presence of rhIL-2, or both rhIL-2 and rhIL-21. The average cell counts from two donors are shown. (c) The mean viability of T cells during co-culture on aAPC and cytokine(s) (n = 2). Percentage expression (n = 2) of (d) CD3 + CD8 + and (e) CD3 + CD4 + T cells upon co-culture with aAPC and cytokines(s). (f) Numeric expansion of T cells measured over culture time on aAPC and rhIL-2 and 21 recorded as the mean inferred cell count (black dashed line) based on 5 healthy subjects (blue solid line) and 5 non-trial canines with NHL (orange solid line). Upward arrows indicate timing for the addition of γ-irradiated OKT3-loaded aAPC.

Article Snippet: T cells obtained directly from PBMC were purified to homogeneity using mouse anti-canine CD3 antibody and a BD FACS Aria II high speed sorter (BD Biosciences) prior to analysis.

Techniques: Isolation, Irradiation, Recombinant, Co-Culture Assay, Genetically Modified, Clone Assay, Expressing, Cell Counting

Journal: Scientific Reports

Article Title: Adoptive T-cell therapy improves treatment of canine non–Hodgkin lymphoma post chemotherapy

doi: 10.1038/srep00249

Figure Lengend Snippet: (a) Ratio of CD4:CD8 T cells in PB after CHOP, but before adoptive transfer of T cells and compared with measurements taken three hours after each T-cell infusion. The grey shaded area represents the mean CD4:CD8 ratio (1.6:1) in healthy subjects (n = 4). (b) Mean T-cell counts in PB from 6 canines before and after adoptive transfer of T cells. The grey shaded area represents the range for CD8 + T cells in healthy subjects (n = 4, 581 to 958 cells/µL). Arrows represent days T cells were infused. (c) Mean expression of CD3 + T cells pre-stained with red fluorescent dye, PKH-26, in the PB of 6 canines. Arrows represent days T cells were infused. (d) Evaluation of fluorescence and staining of CD3 from a LN biopsy. Frozen tissues were viewed with fluorescent microscopy to detect ( top ) PKH-26 + (red) T cells and ( bottom ) co-stained with anti-CD3 (black) to validate T-cell presence.

Article Snippet: T cells obtained directly from PBMC were purified to homogeneity using mouse anti-canine CD3 antibody and a BD FACS Aria II high speed sorter (BD Biosciences) prior to analysis.

Techniques: Adoptive Transfer Assay, Expressing, Staining, Fluorescence, Microscopy

Journal: Scientific Reports

Article Title: Adoptive T-cell therapy improves treatment of canine non–Hodgkin lymphoma post chemotherapy

doi: 10.1038/srep00249

Figure Lengend Snippet: (a) Serum TK concentration in canines (n = 5) plotted with the number of CD3 + CD8 + T cells in PB. Arrows indicate the infusion of T cells. (b) NLR on study day 35 plotted as a function of tumor-free survival. Horizontal arrow indicates NLR in PB of healthy canines. (c) Expression of granzyme B in propagated T cells, assessed at time of cryopreservation, plotted as a function of tumor-free survival.

Article Snippet: T cells obtained directly from PBMC were purified to homogeneity using mouse anti-canine CD3 antibody and a BD FACS Aria II high speed sorter (BD Biosciences) prior to analysis.

Techniques: Concentration Assay, Expressing

Journal: The Journal of Veterinary Medical Science

Article Title: Influence of transfusion of lymphokine-activated T killer cells on inflammatory responses in dogs after laparotomy

doi: 10.1292/jvms.15-0626

Figure Lengend Snippet: Effects of T-LAK therapy on numbers of PBLs and T lymphocyte subsets. (A) PBLs, (B) CD3 + cells, (C) CD4 + cells, (D) CD8 + cells. The black squares and solid line represent the T-LAK group, and the white diamonds and dashed line represent the control group. The numbers of PBLs and all T lymphocyte subsets in the T-LAK group were significantly higher than those in the control group by two-way ANOVA ( P <0.05).

Article Snippet: T-LAK therapy : The bottom of a 250 m l flask for suspension cell culture (Sumitomo Bakelite Co., Ltd., Tokyo, Japan) was coated with 10 m l of 5 μ g/m l anti-canine CD3 antibody solution diluted with Dulbecco’s PBS (D-PBS; Nacalai Tesque).

Techniques: Control