Therapy-induced antitumor vaccination in neuroblastomas by the combined targeting of IL-2 and TNF alpha

L19-IL2 and L19TNF are fusion proteins composed of L19(scFv), specific for the angiogenesis-associated ED-B containing fibronectin isoform and IL-2 or TNF. Because of the tumor targeting properties of L19, IL-2 and TNF concentrate at therapeutic doses at the tumor vascular level. To evaluate the therapeutic effects of L19-IL2 and L19mTNF in neuroblastoma (NB)-bearing mice, A/J mice bearing Neuro2A or NIE115 NB were systemically treated with L19-IL2 and L19mTNF, alone or in combination protocols. Seventy percent of Neuro2A- and 30% of NIE115-bearing mice were cured by the combined treatment with L19-IL2 and L19mTNF, and further rejected a homologous tumor challenge, indicating specific antitumor immune memory. The immunological bases of tumor cure and rejection were studied. A highly efficient priming of CD4+ T helper cells and CD8+ CTL effectors was generated, paralleled by massive infiltration in the tumor tissue of CD4+ and CD8+ T cells at day 16 after tumor cell implantation, when, after therapy, tumor volume was drastically reduced and tumor necrosis reached about 80%. The curative treatment resulted in a long-lasting antitumor immune memory, accompanied by a mixed Th1/Th2 type of response. Concluding, L19-IL2 and L19mTNF efficiently cooperate in determining a high percentage of NB cure that, in our experimental models, is strongly associated to the generation of adaptive immunity involving CD4+ and CD8+ T cells

vandetanib improves anti tumor effects of l19mtnfα in xenograft models of esophageal cancer

Purpose: Targeting the tumor vasculature by vascular disrupting agents (VDAs) has shown therapeutic activity in mouse models. In most cases, however, VDA efficacy is substantially compromised by the inability of these drugs to completely kill tumor cells located at the periphery of the tumor mass. In this study, we investigated anti-tumor effects of L19mTNFα, a fusion protein composed of L19 (scFv), specific for the angiogenesis-associated ED-B containing fibronectin isoform, and murine TNFα, in xenograft models of esophageal cancer. Experimental design: We evaluated ED-B expression in esophageal cancer samples. Subsequently, we generated subcutaneous xenografts from primary tumors, treated them with the L19mTNFα scFv, and determined effects on tumor vasculature, viability and proliferation, and VEGF expression and infiltration by hematopoietic cells. To overcome tumor resistance, L19mTNFα scFv was combined with vandetanib, a tyrosine kinase inhibitor of VEGF receptor, epidermal growth factor receptor, and RET signaling. Results: ED-B was broadly expressed by esophageal cancer cell lines, as well as xenografts and primary surgical samples of esophageal cancer. Administration of L19mTNFα acutely damaged tumor vasculature and increased necrosis, indicating a VDA-like activity of this immunoconjugate. This event was followed, however, by rapid tumor growth recovery associated with increased expression of VEGF and recruitment of CD11b+Gr1+ myeloid cells into tumors. Combination of L19mTNFα with vandetanib severely impaired vascular functions in tumors, leading to a reduction of cell proliferation and increased necrosis, without apparent signs of toxicity. Conclusion: These findings indicate that a combination of vascular damaging agents with anti-angiogenic drugs could represent a promising therapeutic strategy for esophageal cancer. Clin Cancer Res; 17(3); 447–58. ©2010 AACR

L19-IL2 immunocytokine in combination with the anti-syndecan-1 46F2SIP antibody format: A new targeted treatment approach in an ovarian carcinoma model

Epithelial ovarian cancer (EOC) is the fifth most common cancer affecting the female population. At present, different targeted treatment approaches may improve currently employed therapies leading either to the delay of tumor recurrence or to disease stabilization. In this study we show that syndecan-1 (SDC1) and tumor angiogenic-associated B-fibronectin isoform (B-FN) are involved in EOC progression and we describe the prominent role of SDC1 in the vasculogenic mimicry (VM) process. We also investigate a possible employment of L19-IL2, an immunocytokine specific for B-FN, and anti-SDC1 46F2SIP (small immuno protein) antibody in combination therapy in a human ovarian carcinoma model. A tumor growth reduction of 78% was obtained in the 46F2SIP/L19-IL2-treated group compared to the control group. We observed that combined treatment was effective in modulation of epithelial-mesenchymal transition (EMT) markers, loss of stemness properties of tumor cells, and in alleviating hypoxia. These effects correlated with reduction of VM structures in tumors from treated mice. Interestingly, the improved pericyte coverage in vascular structures suggested that combined therapy could be efficacious in induction of vessel normalization. These data could pave the way for a possible use of L19-IL2 combined with 46F2SIP antibody as a novel therapeutic strategy in EOC

Diseño, análisis sensorial y caracterización fisicoquímica de un dip de berenjena ahumada (Solanum Melongena l.)

El dip es un tipo de salsa elaborada a base de varios ingredientes, que suele ser más espesa que las salsas habituales,este se unta en alimentos sólidos como verduras, pan, tostadas, carnes, nachos y otros, complementando las característicassensoriales del alimento y a su vez dando un aporte nutricional. Se realizó un dip de berenjena ahumada(Solanum melongena L) con el objetivo de evaluar la aceptabilidad y sus atributos (Textura, Color, Sabor, Aromay Apariencia); y a su vez determinar un acompañante. Para la elaboración del dip se utilizó una formulación estandarizadateniendo una concentración de berenjena del 45%; mayonesa, un 52%; ajo, 1%; limón, 1,7% y sal 0,3%empleando una estufa para el ahumado y una licuadora casera para la mezcla homogénea de todos los ingredientes,logrado un dip de textura y apariencia deseada. Se realizaron pruebas sensoriales evaluando los atributos con 19consumidores semi-entrenados, también se realizó pruebas físico-químicas obteniendo como resultado 76% de humedadpor gramo de producto, 8 °Brix, pH de 4,1 y 0,2% de ácido málico. Los resultados sensoriales revelaron queel aroma y el sabor fueron los atributos que presentaron menor aceptabilidad con un promedio de 3,26 manifestandopoca afinidad de los catadores con el olor ahumado. En general el dip tuvo una aceptabilidad media de 3,65,donde los acompañantes con mayor propuesta fueron carnes y tostadas con un porcentaje de elección del 35%

Diseño, análisis sensorial y caracterización fisicoquímica de un dip de berenjena ahumada (Solanum Melongena l.)

El dip es un tipo de salsa elaborada a base de varios ingredientes, que suele ser más espesa que las salsas habituales,este se unta en alimentos sólidos como verduras, pan, tostadas, carnes, nachos y otros, complementando las característicassensoriales del alimento y a su vez dando un aporte nutricional. Se realizó un dip de berenjena ahumada(Solanum melongena L) con el objetivo de evaluar la aceptabilidad y sus atributos (Textura, Color, Sabor, Aromay Apariencia); y a su vez determinar un acompañante. Para la elaboración del dip se utilizó una formulación estandarizadateniendo una concentración de berenjena del 45%; mayonesa, un 52%; ajo, 1%; limón, 1,7% y sal 0,3%empleando una estufa para el ahumado y una licuadora casera para la mezcla homogénea de todos los ingredientes,logrado un dip de textura y apariencia deseada. Se realizaron pruebas sensoriales evaluando los atributos con 19consumidores semi-entrenados, también se realizó pruebas físico-químicas obteniendo como resultado 76% de humedadpor gramo de producto, 8 °Brix, pH de 4,1 y 0,2% de ácido málico. Los resultados sensoriales revelaron queel aroma y el sabor fueron los atributos que presentaron menor aceptabilidad con un promedio de 3,26 manifestandopoca afinidad de los catadores con el olor ahumado. En general el dip tuvo una aceptabilidad media de 3,65,donde los acompañantes con mayor propuesta fueron carnes y tostadas con un porcentaje de elección del 35%

Diseño, análisis sensorial y caracterización fisicoquímica de un dip de berenjena ahumada (Solanum Melongena l.)

El dip es un tipo de salsa elaborada a base de varios ingredientes, que suele ser más espesa que las salsas habituales,este se unta en alimentos sólidos como verduras, pan, tostadas, carnes, nachos y otros, complementando las característicassensoriales del alimento y a su vez dando un aporte nutricional. Se realizó un dip de berenjena ahumada(Solanum melongena L) con el objetivo de evaluar la aceptabilidad y sus atributos (Textura, Color, Sabor, Aromay Apariencia); y a su vez determinar un acompañante. Para la elaboración del dip se utilizó una formulación estandarizadateniendo una concentración de berenjena del 45%; mayonesa, un 52%; ajo, 1%; limón, 1,7% y sal 0,3%empleando una estufa para el ahumado y una licuadora casera para la mezcla homogénea de todos los ingredientes,logrado un dip de textura y apariencia deseada. Se realizaron pruebas sensoriales evaluando los atributos con 19consumidores semi-entrenados, también se realizó pruebas físico-químicas obteniendo como resultado 76% de humedadpor gramo de producto, 8 °Brix, pH de 4,1 y 0,2% de ácido málico. Los resultados sensoriales revelaron queel aroma y el sabor fueron los atributos que presentaron menor aceptabilidad con un promedio de 3,26 manifestandopoca afinidad de los catadores con el olor ahumado. En general el dip tuvo una aceptabilidad media de 3,65,donde los acompañantes con mayor propuesta fueron carnes y tostadas con un porcentaje de elección del 35%

Dietary Fat Patterns and Outcomes in Acute Pancreatitis in Spain

Background/Objective: Evidence from basic and clinical studies suggests that unsaturated fatty acids (UFAs) might be relevant mediators of the development of complications in acute pancreatitis (AP). Objective: The aim of this study was to analyze outcomes in patients with AP from regions in Spain with different patterns of dietary fat intake. Materials and Methods: A retrospective analysis was performed with data from 1,655 patients with AP from a Spanish prospective cohort study and regional nutritional data from a Spanish cross-sectional study. Nutritional data considered in the study concern the total lipid consumption, detailing total saturated fatty acids, UFAs and monounsaturated fatty acids (MUFAs) consumption derived from regional data and not from the patient prospective cohort. Two multivariable analysis models were used: (1) a model with the Charlson comorbidity index, sex, alcoholic etiology, and recurrent AP; (2) a model that included these variables plus obesity. Results: In multivariable analysis, patients from regions with high UFA intake had a significantly increased frequency of local complications, persistent organ failure (POF), mortality, and moderate-to-severe disease in the model without obesity and a higher frequency of POF in the model with obesity. Patients from regions with high MUFA intake had significantly more local complications and moderate-to-severe disease; this significance remained for moderate-to-severe disease when obesity was added to the model. Conclusions: Differences in dietary fat patterns could be associated with different outcomes in AP, and dietary fat patterns may be a pre-morbid factor that determines the severity of AP. UFAs, and particulary MUFAs, may influence the pathogenesis of the severity of AP

Optimal MHC-II-restricted tumor antigen presentation to CD4+ T helper cells: the key issue for development of anti-tumor vaccines

Present immunoprevention and immunotherapeutic approaches against cancer suffer from the limitation of being not “sterilizing” procedures, as very poor protection against the tumor is obtained. Thus newly conceived anti-tumor vaccination strategies are urgently needed. In this review we will focus on ways to provide optimal MHC class II-restricted tumor antigen presentation to CD4+ T helper cells as a crucial parameter to get optimal and protective adaptive immune response against tumor. Through the description of successful preventive or therapeutic experimental approaches to vaccinate the host against the tumor we will show that optimal activation of MHC class II-restricted tumor specific CD4+ T helper cells can be achieved in various ways. Interestingly, the success in tumor eradication and/or growth arrest generated by classical therapies such as radiotherapy and chemotherapy in some instances can be re-interpreted on the basis of an adaptive immune response induced by providing suitable access of tumor-associated antigens to MHC class II molecules. Therefore, focussing on strategies to generate better and suitable MHC class II–restricted activation of tumor specific CD4+ T helper cells may have an important impact on fighting and defeating cancer

Synthesis of tenascin and laminin beta2 chain in human bronchial epithelial cells is enhanced by cysteinyl leukotrienes via CysLT1 receptor

<p>Abstract</p> <p>Background</p> <p>Cysteinyl leukotrienes (CysLTs) are key mediators of asthma, but their role in the genesis of airway remodeling is insufficiently understood. Recent evidence suggests that increased expression of tenascin (Tn) and laminin (Ln) β2 chain is indicative of the remodeling activity in asthma, but represents also an example of deposition of extracellular matrix, which affects the airway wall compliance. We tested the hypothesis that CysLTs affect production of Tn and Ln β2 chain by human bronchial epithelial cells and elucidated, which of the CysLT receptors, CysLT₁or CysLT₂, mediate this effect.</p> <p>Methods</p> <p>Cultured BEAS-2B human bronchial epithelial cells were stimulated with leukotriene D₄(LTD₄) and E₄(LTE₄) and evaluated by immunocytochemistry, Western blotting, flow cytometry, and RT-PCR. CysLT receptors were differentially blocked with use of montelukast or BAY u9773.</p> <p>Results</p> <p>LTD₄and LTE₄significantly augmented the expression of Tn, whereas LTD₄, distinctly from LTE₄, was able to increase also the Ln β2 chain. Although the expression of CysLT₂prevailed over that of CysLT₁, the up-regulation of Tn and Ln β2 chain by CysLTs was completely blocked by the CysLT₁-selective antagonist montelukast with no difference between montelukast and the dual antagonist BAY u9773 for the inhibitory capacity.</p> <p>Conclusion</p> <p>These findings suggest that the CysLT-induced up-regulation of Tn and Ln β2 chain, an important epithelium-linked aspect of airway remodeling, is mediated predominantly by the CysLT₁receptor. The results provide a novel aspect to support the use of CysLT₁receptor antagonists in the anti-remodeling treatment of asthma.</p

Particular distribution and expression pattern of endoglin (CD105) in the liver of patients with hepatocellular carcinoma

<p>Abstract</p> <p>Background</p> <p>Endoglin (CD105) has been considered a prognostic marker for hepatocellular carcinoma (HCC), and widely used as an appropriate targeting for antiangenesis therapy in some cancers. Our aim was to evaluate the distribution and expression of CD105 in the liver of patients with HCC, and to discuss whether CD105 may be used as an appropriate targeting for antiangenesis therapy in HCC.</p> <p>Methods</p> <p>Three parts of liver tissues from each of 64 patients with HCC were collected: tumor tissues (TT), adjacent non-tumor (AT) liver tissues within 2 cm, and tumor free tissues (TF) 5 cm far from the tumor edge. Liver samples from 8 patients without liver diseases served as healthy controls (HC). The distribution and expression of CD105 in tissues were evaluated by immunohistochemistry, Western blotting analysis, and real-time PCR. HIF-1alpha and VEGF₁₆₅protein levels in tissues were analyzed by Immunohistochemistry and Western blotting analysis or ELISA.</p> <p>Results</p> <p>CD105 was positively stained mostly in a subset of microvessels 'endothelial sprouts' in TT of all patients while CD105 showed diffuse positive staining, predominantly on hepatic sinus endothelial cells in the surrounding of draining veins in TF and AT. The mean score of MVD-CD105 (mean ± SD/0.74 mm²) was 19.00 ± 9.08 in HC, 153.12 ± 53.26 in TF, 191.12 ± 59.17 in AT, and 85.43 ± 44.71 in TT, respectively. Using a paired <it>t </it>test, the expression of CD105 in AT and TF was higher than in TT at protein (MVD, <it>p </it>= 0.012 and <it>p </it>= 0.007, respectively) and mRNA levels (<it>p </it>< 0.001 and <it>p </it>= 0.009, respectively). Moreover, distribution and expression of CD105 protein were consistent with those of HIF-1alpha and VEGF₁₆₅protein in liver of patients with HCC. The level of <it>CD105 </it>mRNA correlated with VEGF₁₆₅level in TF (r = 0.790, <it>p </it>= 0.002), AT (r = 0.723, <it>p </it>< 0.001), and TT (r = 0.473, <it>p </it>= 0.048), respectively.</p> <p>Conclusion</p> <p>It is demonstrated that CD105 was not only present in neovessels in tumor tissues, but also more abundant in hepatic sinus endothelium in non-tumor tissues with cirrhosis. Therefore, CD105 may not be an appropriate targeting for antiangenesis therapy in HCC, especially with cirrhosis.</p