B and T lymphocyte attenuator (BTLA)-herpesvirus entry mediator (HVEM) signaling coinhibitory pathway is believed to impair antitumor immune competences. An intriguing unresolved question is whether blockade of BTLA-HVEM guides an effective therapeutic tool against established tumors. To address this issue, we constructed a eukaryotic expression plasmid (psBTLA) that expressed the extracellular domain of murine BTLA (soluble form of BTLA), which could bind HVEM, the ligand of BTLA, and block BTLA-HVEM interactions. The data in this study showed that treatment by injection of psBTLA resulted in down-regulation of IL-10 and TGF-β and promotion of dendritic cell function by increasing the expression of B7-1 and IL-12, but the adaptive antitumor immune responses achieved by psBTLA administration alone were limited and could not eradicate the tumor effectively. Next, we evaluated the immunotherapeutic efficacy and mechanism of combination therapy of heat shock protein 70 (HSP70) vaccine/psBTLA by using murine TC-1 cervical cancer mice as an ectopic tumor model. Our in vivo studies revealed that treatment with HSP70 vaccine alone did not lead to satisfactory tumor growth inhibition, whereas cotreatment with psBTLA significantly improved antitumor immunity and compensated the deficiency of HSP70 vaccine by increasing the expression of Th1 cytokines, IL-2, and IFN-γ and decreasing transcription levels of IL-10, TGF-β, and Foxp3 in the tumor microenvironment. Taken together, our findings indicate that blocking the BTLA-HVEM interaction with sBTLA enhances antitumor efficacy and results in a significant synergistic effect against existent tumor cells in vivo when combined with the HSP70 vaccine.

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Human papillomavirus (HPV) infection is the most important risk factor for the development of cervical cancer.The oncogene E7 from high-risk HPV strains has the ability to immortalize epithelial cells and increase cellular transformation in culture. In this study, we explored the possibility of preventing cervical cancer growth by inhibiting HPV16 E7 expression through gene transfer of an antisense construct. A recombinant adeno-associated virus (rAAV) vector was chosen for the transfer, based on its transfection efficiency, in vivo stability, and lack of detectable pathology. In vitro transfer of an rAAV vector expressing antisense HPV16 E7 (AAV-HPV16E7AS) inhibited cell proliferation, induced apoptosis, reduced cell migration, and restrained in vivo proliferation of HPV16/HPV18^positive cervical cancer CaSki cells. These results indicate that down-regulation of HPV16 E7 with antisense RNA is beneficial in reducing the tumorigenicity of CaSki cells, and rAAV vectors ought to be a new efficient approach for delivering the expression of therapeutic genes.Most acquired and inherited diseases are rooted at the genetic level, and the ability to correct genetic defects that cause disease remains the ultimate goal of gene therapy. For some diseases, gene therapy presents the only hope for patients, but truly effective gene therapy continues to be elusive. Although a great variety of genes show potential for the treatment of human disease, delivering genetic material with therapeutic potential to specific target sites remains a challenge (1, 2).Gene delivery systems can be categorized as viral or nonviral systems. The more commonly used viral gene delivery systems are retrovirus (3), adenovirus (4), and adeno-associated virus (AAV; ref. 5) vectors. Commonly used nonviral delivery systems include cationic liposome (6), HVJ-liposome (7), and mechanical approaches, such as ''gene gun' ' (8), DNA infusion, and DNA injection (9). Many gene delivery systems have shown some degree of success in vitro, but all have fallen short in in vivo trials. The major problems encountered have been low efficiency of gene delivery and/or inability to achieve targeted gene placement.Antisense technology has been used mainly to knock out or down-regulate the expression of certain genes associated with disease. Antisense constructs are small and generally do not code for any known biological activities in the host, and in early trials, they have proven to be well tolerated (10, 11). However, the effectiveness of antisense therapy has generally been short lived, and problems with the efficiency and specificity of gene delivery have also limited its use. The side effects that result from nonspecific gene delivery can be circumvented if the chosen target for antisense attack is an acquired genetic material, such as a viral sequence, and not a somatic gene. Targeting acquired genes has been proposed for diseases associated with infectious agents, including some cancers, such as cervical cancer associated with human papillomavirus 16 (HP...

To investigate the role of S100 calcium-binding protein A16 (S100A16) in hepatic lipid metabolism, S100a16 transgenic, S100a16 knockdown, and wildtype C57BL/6 mice were fed either a high-fat diet (HFD) or normal-fat diet (NFD) for 16 weeks. The results showed that for HFD-fed mice, S100a16 transgenic mice showed significantly more severe fatty liver than other HFD-fed mice, with a significant increase in serum triglyceride (TG) concentration, with more and larger lipid droplets in the liver, whereas S100a16 knockdown mice were completely opposite, with liver fat lesions and TG serological changes being the mildest; for NFD-fed mice, liver fat accumulation and serum TG concentrations were significantly lower than those fed HFD, and no significant lipid droplets were found in the liver. Further, we found that calmodulin (CaM) interacts with S100A16, a member of the AMP-activated protein kinase (AMPK) pathway. Our research found that S100A16 regulates the AMPK pathway-associated protein by interacting with CaM to regulate liver lipid synthesis. S100A16 regulates liver lipid metabolism through the CaM/CAMKK2/AMPK pathway. Overexpression of S100A16 promotes the deterioration of fatty liver induced by HFD, and low expression of S100A16 can attenuate fatty liver. K E Y W O R D Scalmodulin, fatty liver, S100 calcium-binding protein A16