Mouse anti-Rat CD80 Monoclonal antibody, clone 4I6

Radiofrequency ablation (RFA) is the most common approach to thermal ablation for cancer therapy. Unfortunately, its efficacy is limited by incomplete ablation, and further optimization of RFA is required. Here, we demonstrate that incubation at 65 degrees C triggers more EG7 tumor cell death by necrosis than treatment at 45 degrees C, and the 65 degrees C-treated cells are more effective at inducing antigen-specific CD8(+) cytotoxic T lymphocyte (CTL) responses after injection in mice than the 45 degrees C-treated ones. Dendritic cells (DCs) that phagocytose 65 degrees C-treated EG7 cells become mature with upregulated MHCII and CD80 expression and are capable of efficiently inducing effector CTLs in mouse tumor models. RFA (65 degrees C) therapy of EG7 tumors induces large areas of tumor necrosis and stimulates CTL responses. This leads to complete regression of small (similar to 100 mm(3)) tumors but fails to suppress the growth of larger (similar to 350 mm(3)) tumors. The administration of the Toll-like receptor-9 (TLR9) agonist unmethylated cytosine-phosphorothioate-guanine oligonucleotide (CpG) to DCs phagocytosing 65 degrees C-treated EG7 cells enhances the expression of MHCII and CD40 on DCs as well as DC-induced stimulation of CTL responses. Importantly, the intratumoral administration of CpG following RFA also increases the frequencies of tumor-associated immunogenic CD11b(-)CD11c(+)CD103(+) DC2 and CD11b(+)F4/80(+)MHCII(+) M1 macrophages and increases CD4(+) and CD8(+) T-cell tumor infiltration, leading to enhanced CD4(+) T cell-dependent CTL responses and potent inhibition of primary RFA-treated or distant untreated tumor growth as well as tumor lung metastasis in mice bearing larger tumors. Overall, our data indicate that CpG administration, which enhances RFA-induced CTL responses and ultimately potentiates the inhibition of primary tumor growth and lung metastasis, is a promising strategy for improving RFA treatment, which may assist in optimizing this important cancer therapy.

Microglia are first-line defense antigen-presenting phagocytes in the central nervous system. Activated microglial cells release pro-inflammatory cytokines and can trigger an oxidative burst. The amino acid glycine exerts anti-inflammatory, immunomodulatory and cytoprotective effects and influences cell volume regulation. This study aimed to investigate the role of glycine in the modulation of inflammatory processes in mouse BV-2 microglial cells. Inflammatory stress was induced by lipopolysaccharide/interferon-gamma (LPS/IFN-gamma) treatment for 24 h in the absence or presence of 1 or 5 mM glycine. Cells were analyzed by flow cytometry for cell volume, side scatter, apoptosis/necrosis and expression of activation-specific surface markers. Apoptosis progression was monitored by life cell imaging. Reduced glutathione/oxidized glutathione (GSH/GSSG) ratios and release of the pro-inflammatory cytokines IL-6 and TNF-alpha were measured using luminescence-based assays and ELISA, respectively. We found that LPS/IFN-gamma-induced apoptosis was decreased and the fraction of living cells was increased by glycine. Expression of the surface markers CD11b, CD54 and CD80 was dose-dependently increased, while IL-6 and TNF-alpha release was not altered compared to LPS/IFN-gamma-treated cells. We showed that in BV-2 microglial cells glycine improves viability and counteracts deleterious responses to LPS/IFN-gamma, which might be relevant in neurodegenerative processes associated with inflammation, like Alzheimer's or Parkinson's disease.