Oligodeoxynucleotides expressing polyguanosine motifs promote antitumor activity through the upregulation of IL-2.

The primary goal of cancer immunotherapy is to elicit an immune response capable of eliminating the tumor. One approach toward accomplishing that goal uses general (rather than tumor-specific) immunomodulatory agents to boost the number and activity of pre-existing CTLs. We find that the intratumoral injection of polyguanosine (poly-G) oligonucleotides (ODN) has such an effect, boosting antitumor immunity and promoting tumor regression. The antitumor activity of poly-G ODN was mediated through CD8 T cells in a TLR9-independent manner. Mechanistically, poly-G ODN directly induced the phosphorylation of Lck (an essential element of the T cell-signaling pathway), thereby enhancing the production of IL-2 and CD8 T cell proliferation. These findings establish poly-G ODN as a novel type of cancer immunotherapy.

Beclin1-induced autophagy abrogates radioresistance of lung cancer cells by suppressing osteopontin.

Osteopontin (OPN) serves as an indicator of resistance to radiotherapy. However, the role of OPN in the development of acquired radioresistance in human lung cancer cells has not yet been fully elucidated. Therefore, the potential importance of OPN as a marker of lung cancer with a potential significant role in the development of radioresistance against repeated radiotherapy has prompted us to define the pathways by which OPN regulates lung cancer cell growth. In addition, autophagy has been reported to play a key role in the radiosensitization of cancer cells. Here, we report that increased OPN expression through induction of nuclear p53 following irradiation was inhibited by exogenous beclin-1 (BECN1). Our results clearly show that BECN1 gene expression led to induction of autophagy and inhibition of cancer cell growth and angiogenesis. Our results suggest that the induction of autophagy abrogated the radioresistance of the cancer cells. Interestingly, we showed that knockdown of OPN by lentivirus-mediated shRNA induced the autophagy of human lung cancer cell. Taken together, these results suggest that OPN and BECN1 can be molecular targets for overcoming radioresistance by controlling autophagy.

Potential of transfected muscle cells to contribute to DNA vaccine immunogenicity.

The mechanism(s) by which DNA vaccines trigger the activation of Ag-specific T cells is incompletely understood. A series of in vivo and in vitro experiments indicates plasmid transfection stimulates muscle cells to up-regulate expression of MHC class I and costimulatory molecules and to produce multiple cytokines and chemokines. Transfected muscle cells gain the ability to directly present Ag to CD8 T cells through an IFN-regulatory factor 3-dependent process. These findings suggest that transfected muscle cells at the site of DNA vaccination may contribute to the magnitude and/or duration of the immune response initiated by professional APCs.

Diffusion of bioactive molecules through the walls of the medial tissue-engineered hybrid ePTFE grafts for applications in designs of vascular tissue regeneration.

Strategies of better vascular tissue engineering may require delivery of soluble bioactive signals in cell culture medium to the cells in tissue-regenerating constructs. We measured the diffusivity and permeability of model tissue-engineering bioactive molecules such as water and heparin through the walls of both a hybrid ePTFE graft and a porcine carotid artery, a model vascular tissue. While diffusivities of H(3)-water and H(3)-heparin were measured as 3.9 x 10(-) (6) and 1.6 x 10(-) (6) cm(2)/s in the artery, respectively, under diffusional circulation of cell culture medium through the lumens of the carotid arteries, their corresponding permeabilities were 4.7 x 10(-) (5) and 2.0 x 10(-) (5) cm/s. On the other hand, diffusivities of H(3)-water and H(3)-heparin were also measured as 5.1 x 10(-) (6) and 4.7 x 10(-) (6) cm(2)/s, respectively, in the tissue-engineered hybrid ePTFE grafts; their corresponding permeabilities were 5.1 x 10(-) (5) and 3.7 x 10(-) (5) cm/s. The hybrid graft tissues were engineered by replacing the biodegradable, porous poly(lactide-co-glycolide) layers coated on the ePTFE surfaces with smooth muscle cell-derived tissues for 6 weeks. We analyzed the morphologies of the artery and the engineered hybrid ePTFE tissues with scanning electron microscopy and H&E stains. While the artery had its typical structure properties with layers of intima, media and adventitia, the tissue-engineered ePTFE hybrid graft had two layers of engineered tissues on the inner and outer surfaces of the ePTFE. There were no significant differences among the luminal tissue morphologies of the test samples from the effects of diffusion flow applications, with minor changes on their luminal surfaces. The results of water and heparin diffusion experiments indicated that these bioactive molecules were well transported from the cell culture medium to the tissue-engineering cells, enough to support tissue regeneration. We hope that these transport results may elucidate the transport behaviors of soluble nutrient molecules and biological signals through the vascular constructs under tissue engineering processes.