Dual activation of Toll-like receptors 7 and 9 impairs the efficacy of antitumor vaccines in murine models of metastatic breast cancer.

PURPOSE: Since combination of Toll-like receptor (TLR) ligands could boost antitumor immunity, we evaluated the efficacy of dendritic cell (DC) vaccines upon dual activation of TLR9 and TLR7 in breast cancer models. METHODS: DCs were generated from mouse bone marrow or peripheral blood from healthy human donors and stimulated with CpG1826 (mouse TLR9 agonist), CpG2006 or IMT504 (human TLR9 agonists) and R848 (TLR7 agonist). Efficacy of antitumor vaccines was evaluated in BALB/c mice bearing metastatic mammary adenocarcinomas. RESULTS: CpG-DCs improved the survival of tumor-bearing mice, reduced the development of lung metastases and generated immunological memory. However, dual activation of TLRs impaired the efficacy of DC vaccines. In vitro, we found that R848 inhibited CpG-mediated maturation of murine DCs. A positive feedback loop in TLR9 mRNA expression was observed upon CpG stimulation that was inhibited in the presence of R848. Impaired activation of NF-κB was detected when TLR9 and TLR7 were simultaneously activated. Blockade of nitric oxide synthase (NOS) and indoleamine-pyrrole-2,3-dioxygenase (IDO) improved the activation of CpG-DCs. When we evaluated the effect of combined activation of TLR9 and TLR7 in human DCs, we found that R848 induced robust DC activation that was inhibited by TLR9 agonists. CONCLUSIONS: These observations provide insight in the biology of TLR9 and TLR7 crosstalk and suggest caution in the selection of agonists for multiple TLR stimulation. Blockade of NOS and IDO could improve the maturation of antitumor DC vaccines. R848 could prove a useful adjuvant for DC vaccines in human patients.

Gene therapy for pituitary tumors.

Pituitary tumors are the most common primary intracranial neoplasms. Although most pituitary tumors are considered typically benign, others can cause severe and progressive disease. The principal aims of pituitary tumor treatment are the elimination or reduction of the tumor mass, normalization of hormone secretion and preservation of remaining pituitary function. In spite of major advances in the therapy of pituitary tumors, for some of the most difficult tumors, current therapies that include medical, surgical and radiotherapeutic methods are often unsatisfactory and there is a need to develop new treatment strategies. Gene therapy, which uses nucleic acids as drugs, has emerged as an attractive therapeutic option for the treatment of pituitary tumors that do not respond to classical treatment strategies if the patients become intolerant to the therapy. The development of animal models for pituitary tumors and hormone hypersecretion has proven to be critical for the implementation of novel treatment strategies and gene therapy approaches. Preclinical trials using several gene therapy approaches for the treatment of anterior pituitary diseases have been successfully implemented. Several issues need to be addressed before clinical implementation becomes a reality, including the development of more effective and safer viral vectors, uncovering novel therapeutic targets and development of targeted expression of therapeutic transgenes. With the development of efficient gene delivery vectors allowing long-term transgene expression with minimal toxicity, gene therapy will become one of the most promising approaches for treating pituitary adenomas.

Corticotropin-releasing (CRH) activity of thymic peptides on CRH-insensitive corticotropic tumor cells

Se sabe que ciertas preparaciones tímicas son capaces de estimular la liberación de corticotrofina (ACTH) en células hipofisarias. No resulta claro, sin embargo, si este efecto es mediado por el receptor para la hormona liberadora de ACTH (CRH). En el presente estudio se observó que la timosina fracción cinco (TF5), el péptido tímico MB-35 y posiblemente ciertas histonas de timo de ternera son capaces de estimular la liberación de ACTH en una sublínea insensible al CRH derivada de la línea tumoral coricotropa AtT20 y por lo tanto denominada AtT20(Cl). El rango de concentraciones efectivas en el cual TF5 y MB-35 mostraron un efecto dosis-dependiente sobre la liberación de ACTH fue de 100 a 2000 µg/ml y de 10 a 100 mg/ml para TF5 y MB-35, respectivamente. Aunque ninguna de estas preparaciones indujo cambios significativos en el contenido intracelular de ACTH en las células AtT20(Cl), se observó una tendencia hacia la depleción a las dosis más altas de RF5. Los resultados obtenidos sugieren que ciertos péptidos tímicos son capaces de estimular la liberación de ACTH en células corticotropas por un mecanismo independiente del receptor par CRH

Corticotropin-releasing (CRH) activity of thymic peptides on CRH-insensitive corticotropic tumor cells

Se sabe que ciertas preparaciones tímicas son capaces de estimular la liberación de corticotrofina (ACTH) en células hipofisarias. No resulta claro, sin embargo, si este efecto es mediado por el receptor para la hormona liberadora de ACTH (CRH). En el presente estudio se observó que la timosina fracción cinco (TF5), el péptido tímico MB-35 y posiblemente ciertas histonas de timo de ternera son capaces de estimular la liberación de ACTH en una sublínea insensible al CRH derivada de la línea tumoral coricotropa AtT20 y por lo tanto denominada AtT20(Cl). El rango de concentraciones efectivas en el cual TF5 y MB-35 mostraron un efecto dosis-dependiente sobre la liberación de ACTH fue de 100 a 2000 Ag/ml y de 10 a 100 mg/ml para TF5 y MB-35, respectivamente. Aunque ninguna de estas preparaciones indujo cambios significativos en el contenido intracelular de ACTH en las células AtT20(Cl), se observó una tendencia hacia la depleción a las dosis más altas de RF5. Los resultados obtenidos sugieren que ciertos péptidos tímicos son capaces de estimular la liberación de ACTH en células corticotropas por un mecanismo independiente del receptor par CRH (AU)