Journal:

Article Title: Dendritic cell maturation enhances CD8 + T-cell responses to exogenous antigen via a proteasome-independent mechanism of major histocompatibility complex class I loading

doi: 10.1046/j.1365-2567.2003.01664.x

Figure Lengend Snippet: Antigen dose dependence of CD8+ T-cell responses to ISCOMS-associated antigen. DC were pulsed with 0–10 μg/ml OVA ISCOMS with or without LPS activation, washed and co-cultured with OT-1 lymphocytes. (a,b) The expression of CD69 and CD25 on CD8+ Vα2+ T cells was assessed after 24 hr in culture, and (c) IFN-γ production was assessed after 48 hr in culture. Results shown are mean ±1 SD for triplicate cultures. (*P < 0·02; **P < 0·01 versus OVA ISCOMS pulse only).

Article Snippet: As described previously, 13 interferon-γ (IFN-γ) levels were determined using a sandwich enzyme-linked immunosorbent assay, with a pair of IFN-γ-specific antibodies (PharMingen) and standardized with known concentrations of recombinant murine IFN-γ (PharMingen).

Techniques: Activation Assay, Cell Culture, Expressing

Journal: Journal of Translational Medicine

Article Title: Therapeutic T cells induce tumor-directed chemotaxis of innate immune cells through tumor-specific secretion of chemokines and stimulation of B16BL6 melanoma to secrete chemokines

doi: 10.1186/1479-5876-5-56

Figure Lengend Snippet: a) mRNA-expression of KC, MCP-1, IP-10, RANTES and MIG in unstimulated and stimulated D5 tumor cells. D5 tumor cells were cultured alone or stimulated with IFN-γ (2, 10, 20 ng/ml) or TNF-α (3.5, 35, 350 U/ml) for 2 and 12 hours. Tumor cells were harvested and mRNA expression determined by RT-PCR. b) Chemokine mRNA-expression of MIP-1α and MIP-1β in unstimulated and stimulated D5 tumor cells. D5 tumor cells were cultured alone or stimulated with IFN-γ (20 ng/ml) and TNF-α (350 U/ml) for 2 and 12 hours. D5 tumor cells were harvested and mRNA expression determined by RT-PCR. RNA isolated from the macrophage cell line J774 was stimulated with IFNγ (20 ng/ml) and LPS (10 μg/ml) for 4 h and used as a positive control. Negative controls included water instead of cDNA.

Article Snippet: DJ2PM cells (10 6 ) were incubated for 24 h in CM media alone or in CM media containing 20 ng recombinant murine IFN-γ or 20 ng recombinant murine TNF-α (both from B&D Biosciences, PharMingen, San-Diego, CA), a combination of both or supernatants of 24 h anti-CD3 stimulated wt effector T cells.

Techniques: Expressing, Cell Culture, Reverse Transcription Polymerase Chain Reaction, Isolation, Positive Control

Journal: Journal of Translational Medicine

Article Title: Therapeutic T cells induce tumor-directed chemotaxis of innate immune cells through tumor-specific secretion of chemokines and stimulation of B16BL6 melanoma to secrete chemokines

doi: 10.1186/1479-5876-5-56

Figure Lengend Snippet: D5 tumor cells were cultured with TNF-α (350 U/ml) or IFN-γ (20 ng/ml) for 24 hours and culture supernatants tested for secretion of KC and MCP-1 by ELISA (mean of 2 experiments).

Article Snippet: DJ2PM cells (10 6 ) were incubated for 24 h in CM media alone or in CM media containing 20 ng recombinant murine IFN-γ or 20 ng recombinant murine TNF-α (both from B&D Biosciences, PharMingen, San-Diego, CA), a combination of both or supernatants of 24 h anti-CD3 stimulated wt effector T cells.

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Journal of Translational Medicine

Article Title: Therapeutic T cells induce tumor-directed chemotaxis of innate immune cells through tumor-specific secretion of chemokines and stimulation of B16BL6 melanoma to secrete chemokines

doi: 10.1186/1479-5876-5-56

Figure Lengend Snippet: a) CCR1 and CCR5 mRNA-expression in unstimulated and stimulated DJ2PM macrophages. DJ2PM macrophages were cultured alone or stimulated with IFN-γ (20 ng/ml) or TNF-α (35 U/ml) and mRNA expression determined by RT-PCR. b) Mean percentage of DJ2PM cells expressing CCR5 receptor as determined by flow cytometric analyses. Cells were stained with specific mAb or isotype control. Graph shows the percentage of PE-TR positive DJ2PM cells (mean ± SEM). Histograms with * exhibit significantly (*p = 0.01) greater expression of CCR5 compared to isotype controls. Data are a summary of 3 consecutive experiments.

Article Snippet: DJ2PM cells (10 6 ) were incubated for 24 h in CM media alone or in CM media containing 20 ng recombinant murine IFN-γ or 20 ng recombinant murine TNF-α (both from B&D Biosciences, PharMingen, San-Diego, CA), a combination of both or supernatants of 24 h anti-CD3 stimulated wt effector T cells.

Techniques: Expressing, Cell Culture, Reverse Transcription Polymerase Chain Reaction, Staining