We have investigated whether poly(I:C) Toll-like receptor 3 (TLR3) and resiquimod Toll-like receptor 7 (TLR7) agonists can serve as vaccine adjuvants and promote the efficiency of therapeutic DNA vaccination against tumors expressing the human papilloma virus 16 (HPV-16) E7 protein. For this purpose, C57BL/6 mice were inoculated with 2 × 10(5) TC-1 cells, and they were then immunized with HPV-16 E7 DNA vaccine alone or with 50 μg of resiquimod or poly(I:C) individually. We found that poly(I:C) and resiquimod could induce more antigen-specific lymphocyte proliferation and cytolytic activity compared to vaccination with E7 DNA alone. While E7 DNA had no significant inhibitory effect on tumor growth, co-administration of poly(I:C) and resiquimod with E7 DNA induced significant tumor regression. Peripheral and local cytokine assays demonstrated that co-administration of poly(I:C) and resiquimod with E7 DNA induced circulating antigen-specific IFN-γ and nonspecific intratumoral IL-12. TLR3 and TLR7 agonists can be used to enhance the immune response to DNA vaccine immunogens. Taken together, these data indicate that combined vaccination with DNA encoding HPV-16 E7 plus TLR agonists provides a strategy for improving the efficacy of a vaccine as a possible immunotherapeutic strategy for cervical cancer.

The release of inflammatory mediators following cortical spreading depression (CSD) is suggested to play a role in pathophysiology of CSD-related neurological disorders. Toll-like receptors (TLR) are master regulators of innate immune function and involved in the activation of inflammatory responses in the brain. TLR3 agonist poly I:C exerts anti-inflammatory effect and prevents cell injury in the brain. The aim of the present study was to examine the effect of systemic administration of poly I:C on the release of cytokines (TNF-α, IFN-γ, IL-4, TGF-β1, and GM-CSF) in the brain and spleen, splenic lymphocyte proliferation, expression of GAD65, GABAAα, GABAAβ as well as Hsp70, and production of dark neurons after induction of repetitive CSD in juvenile rats. Poly I:C significantly attenuated CSD-induced production of TNF-α and IFN-γ in the brain as well as TNF-α and IL-4 in the spleen. Poly I:C did not affect enhancement of splenic lymphocyte proliferation after CSD. Administration of poly I:C increased expression of GABAAα, GABAAβ as well as Hsp70 and decreased expression of GAD65 in the entorhinal cortex compared to CSD-treated tissues. In addition, poly I:C significantly prevented production of CSD-induced dark neurons. The data indicate neuroprotective and anti-inflammatory effects of TLR3 activation on CSD-induced neuroinflammation. Targeting TLR3 may provide a novel strategy for developing new treatments for CSD-related neurological disorders.

BackgroundCervical cancer is the second-most-common cause of malignancies in women worldwide, and the oncogenic activity of the human papilloma virus types (HPV) E7 protein has a crucial role in anogenital tumors. In this study, we have designed a therapeutic vaccine based on chitosan nanodelivery systems to deliver HPV-16 E7 DNA vaccine, considered as a tumor specific antigen for immunotherapy of HPV-associated cervical cancer. We have developed a Nano-chitosan (NCS) as a carrier system for intramuscular administration using a recombinant DNA vaccine expressing HPV-16 E7 (NCS-DNA E7 vaccine). NCS were characterized in vitro for their gene transfection ability.ResultsThe transfection of CS-pEGFP NPs was efficient in CHO cells and the expression of green fluorescent proteins was well observed. In addition, NCS-DNA E7 vaccine induced the strongest E7-specific CD8+ T cell and interferon γ responses in C57BL/6 mice. Mice vaccinated with NCS-DNA E7 vaccine were able to generate potent protective and therapeutic antitumor effects against challenge with E7-expressing tumor cell line, TC-1.ConclusionsThe strong therapeutic effect induced by the Chitosan-based nanodelivery suggest that nanoparticles may be an efficient carrier to improve the immunogenicity of DNA vaccination upon intramuscular administration and the platform could be further exploited as a potential cancer vaccine candidate in humans.

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Introduction: Spreading depression (SD), discovered by Leao in 1944, is a pathophysiological depolarization wave that propagates slowly in the brain (3 mm/min) and causes dramatic ionic and hemodynamic changes. SD appears to act through several mechanisms and receptors, which have not completely understood. Here, we studied the effect of inhibitory system in animal model of SD using immunohistochemistry technique. Materials and Methods: After implanting recording electrodes and cannula over the brain, repetitive SD was induced by KCl injection (2 M) in juvenile rats for four consecutive weeks. Then all rats' brains removed. The mean number of dark neurons in the entorhinal cortex (EC) were determined using toluidine blue staining. To identify the prevalence and distribution of GABAA subunit receptors as well as glutamic acid decarboxylase (GAD), the GABA biosynthetic enzyme, immunohistochemistry technique was performed. Results: The mean number of SD induced was statistically increased during four weeks of experiments. The mean number of dark neurons in EC was significantly increased in SD group compared to sham group. Furthermore, expression of GAD65 in EC significantly increased in SD group compared to sham group. However, both GABA-A α and GABA-A β subunit receptors didn't significantly change in that region after SD. Conclusion:These data suggest that SD is able to damage the neuronal cells in neural tissues in juvenile rats and parallelly lead to enhancement of GAD65 in the central nervous system.

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The ability to induce pluripotency in somatic cells by reprogramming factors offers new opportunities for drug discovery and cell therapy. Induced pluripotent stem cells have the potential to differentiate to various cell types, such as neural and glial cells. Astrocytes, the major glial cells of the central nervous system, play an important role in the function of the brain by regulating of extracellular ions and neurotransmitters, feeding and protection of neurons as well as modulating the activity of microglia. Microglia over-activation can be resulted in brain inflammation with subsequent susceptibility to epileptic seizures. Hypothesis: For many years, embryonic Stem cell transplantation has been examined to prevent seizure attacks in epilepsy. These studies have indicated that adult cells from patient have the ability to be transformed to embryonic stage and convert to a pluripotent stem cell by using some Transcription factors (such as Oct4, Sox2, Nanog, Rex1, Klf, c-Myc and LIN28). Accordingly, fibroblasts from an epileptic have also been reprogrammed to embryonic stage. The resulting iPS cells are isogenic to patient and are able to transform to neurons or glia in a suitable culture condition. Previous studies on ES cell therapy have focused more on neurons than astrocytes. Astrocytes, by secretion of glial cell-derived neurotrophic factor, not only regulate the different microglial activities, such as proliferation, migration and cell adhesion, but can also reduce destructive effects of microglia. Conclusion: In this hypothesis, we suggest a reprogramming system for generating functional astrocyte from human pluripotent stem cell in the presence of neural growth factors. We hypothesize that these cells might reduce neuroinflammation induced by microglia and subsequent susceptibility to seizure. The reprogrammed cells could be used in cell replacement therapy of epilepsy.