RESUMEN
Objective: To improve dendritic cells (DCs) function, we targeted DCs to over express CD40 and inducible costimulator ligand (ICOSL) costimulatory molecules along with total messenger RNA (mRNA) of tumor cells to achieve a safe and effective system for treatment of tumor. Materials and methods: We generated CD40 and ICOSL mRNA in vitro and manipulated DCs using chitosan nanoparticles and also lipofectamine transfection system then examined in vitro and in vivo. Results: Mice bone marrow derived DCs pulsed with total tumor mRNA/CD40 mRNA or ICOSL mRNA showed higher expression of DCs maturation markers (CD40, ICOSL, CD86, and MHC-II) and accelerated secretion of pro-inflammatory cytokines. Co-culture of DCs with T cells enhanced proliferation of T cells and shift toward stronger Th1 cytokine responses especially in presence of CD40 over expressed DCs. Intra-tumor administration of manipulated DCs to 4T1 tumor mice model showed delay in growth of tumor volume, trend to increase in mice survival, and stronger anti-tumor cytokines production in splenocytes of mice model (with higher efficacy of mRNA/chitosan nanoparticle system). Conclusions: Hence, we suggest that targeting intra-tumor DCs to elicit expression of CD40 and ICOSL and present broad range of tumor antigens could yield effective anti-tumor responses. In this regard, CD40 molecule manipulation trigger stronger functions, while mRNA/chitosan nanoparticles system could provide a high potent tool for targeting strategies.
Asunto(s)
Antígenos CD40/genética , Quitosano/química , Células Dendríticas/inmunología , Ligando Coestimulador de Linfocitos T Inducibles/genética , Nanopartículas/química , Neoplasias/inmunología , ARN Mensajero/genética , Animales , Línea Celular Tumoral , Proliferación Celular/genética , Células Cultivadas , Citocinas/inmunología , Citocinas/metabolismo , Células Dendríticas/metabolismo , Femenino , Inmunoterapia/métodos , Activación de Linfocitos/genética , Activación de Linfocitos/inmunología , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Neoplasias/genética , Análisis de Supervivencia , Linfocitos T/inmunología , TransfecciónRESUMEN
BACKGROUND: PIWIL2, a member of Argonaute family of proteins, is exclusively expressed in testis and functions in development and maintenance of germline stem cells. Recently, ectopic expression of PIWIL2 has been reported in a variety of human and mouse tumors. To investigate a potential involvement of PIWIL2 in human bladder cancer, we examined its expression in several human bladder cancer cell lines, normal uroepithelial cell cultures, and some bladder tissues. METHODS: Relative expression of PIWIL2 was determined by real-time quantitative RT-PCR in fifteen bladder carcinoma cell lines, six normal uroepithelial cell cultures and seventy tissue specimens of tumor, tumor margins and morphologically normal tissues of bladder. Specific primers for PIWIL2, TBP and GAPDH (as two internal controls) were used for reverse transcription polymerase chain reaction technique. RESULTS: Real-time qRT-PCR demonstrated high PIWIL2 expression in testis tissue, but at least 240-fold lower expression in all examined cell lines. The highest expression outside testis was observed in one of six primary cultures of normal uroepithelial cells, but even lower expression of PIWIL2 was detected in any of the examined tumor and non-tumor tissues. CONCLUSION: Lack of PIWIL2 expression in most tissues along with its aberrant expression in some tumors candidate the gene as an attractive tumor marker for some neoplasms. However, our study indicates that PIWIL2 does not play a role in carcinogenesis of human bladder carcinoma.