Selma Benito-Martínez

Bim-mediated apoptosis and PD-1/PD-L1 pathway impair reactivity of PD1+/CD127− HCV-specific CD8+ cells targeting the virus in chronic hepatitis C virus infection

PD-1 molecule promotes anergy and IL-7 receptor (CD127) induces an anti-apoptotic effect on T cells. Correlation between PD-1/CD127 phenotype and hepatitis C virus (HCV)-specific CD8(+) cell reactivity in resolved infection (RI) after treatment and persistent HCV-infection (PI) was analysed. Directly ex vivo, PD-1 and CD127 expression on HCV-specific CD8(+) cells displayed a positive and negative correlation, respectively with viraemia. Proliferation after stimulation on PD-1(-)/CD127(+) cells from RI cases was preserved, while it was impaired on PD-1(+)/CD127(-) cells from PI patients. PD1(+)/CD127(+) population was observed in PI, and these maintained expansion ability but they did not target the virus. Frequency of PI cases with HCV-specific CD8(+) cell proliferation increased after anti-PD-L1 and anti-apoptotic treatment. Bim expression on HCV-specific CD8(+) cells from PI patients was enhanced. In conclusion, during chronic HCV infection non-reactive HCV-specific CD8(+) cells targeting the virus are PD-1(+)/CD127(-)/Bim(+) and, blocking apoptosis and PD-1/PD-L1 pathway on them enhances in vitro reactivity.

Medicina herbal china ofertada en páginas web en español: calidad de la información y riesgos

OBJECTIVE The growing use of purchase online via Internet retailers favours the access to potentially toxic natural products. It also contributes to the quick dissemination of the claims made by the retailers on efficacy and safety, these claims being not always based upon reliable information. Here, we have conducted an online search to find Spanish-language retail websites for Chinese herbal medicine and we have analysed them for the quality of product information and the potential health risks. METHODS i) Online search in Google España to find Spanish-language retail websites for Chinese herbal medicine in which we analysed both the claims regarding possible health benefits and adequate safe use indications ii) Identification of potentially toxic herbs in the websites iii) Quantification of Chinese herbal medicines withdrawn by the Agencia Española de Medicamentos y Productos Sanitarios (AEMPS). RESULTS 1) Only one third of the 30 Spanish-language retail websites found which sell Chinese herbal medicine observe the law, given that the other websites include illegal Western disease claims as marketing tools, 2) Five websites provide some safety information, 3) Two websites offer potentially toxic herbs and 4) Chinese herbal medicine adulterated with sibutramine, silfenafil or their analogues make a considerable percentage of the total products withdrawn by the AEMPS. CONCLUSION Online health seekers should be warned about misinformation on retail websites for Chinese herbal medicine and directed to a Spanish government Web site for guidance in safely navigating the Internet for buying Chinese herbal medicine.

Information quality and health risks in Spanish-language retail websites for Chinese herbal medicine

OBJECTIVE The growing use of purchase online via Internet retailers favours the access to potentially toxic natural products. It also contributes to the quick dissemination of the claims made by the retailers on efficacy and safety, these claims being not always based upon reliable information. Here, we have conducted an online search to find Spanish-language retail websites for Chinese herbal medicine and we have analysed them for the quality of product information and the potential health risks. METHODS i) Online search in Google España to find Spanish-language retail websites for Chinese herbal medicine in which we analysed both the claims regarding possible health benefits and adequate safe use indications ii) Identification of potentially toxic herbs in the websites iii) Quantification of Chinese herbal medicines withdrawn by the Agencia Española de Medicamentos y Productos Sanitarios (AEMPS). RESULTS 1) Only one third of the 30 Spanish-language retail websites found which sell Chinese herbal medicine observe the law, given that the other websites include illegal Western disease claims as marketing tools, 2) Five websites provide some safety information, 3) Two websites offer potentially toxic herbs and 4) Chinese herbal medicine adulterated with sibutramine, silfenafil or their analogues make a considerable percentage of the total products withdrawn by the AEMPS. CONCLUSION Online health seekers should be warned about misinformation on retail websites for Chinese herbal medicine and directed to a Spanish government Web site for guidance in safely navigating the Internet for buying Chinese herbal medicine.

OriginalMedicina herbal china ofertada en páginas web en español: calidad de la información y riesgosInformation quality and health risks in Spanish-language retail websites for Chinese herbal medicine

OBJECTIVE The growing use of purchase online via Internet retailers favours the access to potentially toxic natural products. It also contributes to the quick dissemination of the claims made by the retailers on efficacy and safety, these claims being not always based upon reliable information. Here, we have conducted an online search to find Spanish-language retail websites for Chinese herbal medicine and we have analysed them for the quality of product information and the potential health risks. METHODS i) Online search in Google España to find Spanish-language retail websites for Chinese herbal medicine in which we analysed both the claims regarding possible health benefits and adequate safe use indications ii) Identification of potentially toxic herbs in the websites iii) Quantification of Chinese herbal medicines withdrawn by the Agencia Española de Medicamentos y Productos Sanitarios (AEMPS). RESULTS 1) Only one third of the 30 Spanish-language retail websites found which sell Chinese herbal medicine observe the law, given that the other websites include illegal Western disease claims as marketing tools, 2) Five websites provide some safety information, 3) Two websites offer potentially toxic herbs and 4) Chinese herbal medicine adulterated with sibutramine, silfenafil or their analogues make a considerable percentage of the total products withdrawn by the AEMPS. CONCLUSION Online health seekers should be warned about misinformation on retail websites for Chinese herbal medicine and directed to a Spanish government Web site for guidance in safely navigating the Internet for buying Chinese herbal medicine.

Su1649 Dynamics of Mitochondrial Network in Helicobacter pylori Infection

INTRODUCTION: Mitochondria are dynamic organelles that move around and can fuse with one another or can break off from these unions in a continuous process called fusion/ fission. The fusion/fission balance determines the mitochondrial connectivity degree and the mitochondrial network morphology inside the cells. It is known that a fusion/fission imbalance is involved in several pathological processes that occur as a result of an increased cellular death by apoptosis, as it is observed in Helicobacter pylori (HP)-infection. AIMS: To evaluate in gastric epithelial cells the effect of HP on mitochondrial fission/fusion modulation and apoptosis. Relationship with oxidative stress. MATERIALS AND METHODS: Human gastric epithelial cells (AGS, ATCC CRL-1739) were cultured with a HP strain (ATCC51932) for 24h at 108CFU/mL, with and without VitE (10-4M). It was determined: -Cellular (ROS) and mitochondrial (O2.-) oxidative stress using H2DCFDA and MitoSOX Red probes, respectively. -Mitochondrial network phenotype, visualized with NAO staining. -mRNA levels of mitofusin-2 (fusion-involved protein) and drp-1 (dynamin family protein, related to mitochondrial fission). -Number of apoptotic cells visualized with Hoechst dye. Technologies used: Flow Cytometry (FACScan, Becton Dickson), Confocal Microscopy (Olympus FV 1000) and Real Time-PCR (ABI Prism 7000, Applied Biosystems). RESULTS: HP caused an increase of H2DCFDA and MitoSOX fluorescences (2.5 and 1.8-fold, respectively) in HPinfected AGS cells. The reticulotubular phenotype, characteristic of viable cells and composed by long chains of fusedmitochondria, was transformed into a punctiform phenotype (rounded and isolated mitochondria) typical of an increased fission. A 13.6-fold increase in drp-1/ mitofusin-2 mRNA ratio it was observed. Hoechst showed a homogeneous staining in controls, whereas a 30-40% of infected cells displayed a phenotype related to apoptotic features (condensation, peripheral location and shape resembling a half-moon of nuclear chromatin). VitE pre-treatment avoided all these alterations, with statistically significant differences. CONCLUSIONS: The increased oxidative stress in HP-infected gastric epithelial cells is associated with fission-mediated fragmentation of mitochondrial network. The recovery of fusion/fission balance by the antioxidant Vit.E, was accompanied by the inhibition of apoptosis. So, these data could indicate that the mitochondrial dynamic is other stage in the apoptotic cascade triggered by oxidative stress and therefore, it is involved in the development of gastric pathologies related to HP-infection.

Pd1(+) Regulatory T Cells Are Increased in Helicobacter pylori-Associated Gastritis

INTRODUCTION: Regulatory T cells (Treg) (CD4+CD25+FoxP3+) play a fundamental role in modulating the balance between inflammation and immune tolerance and are identified as a factor that contributes to bacterial persistence and to infection chronicity. It is emerging that Treg include different functional and phenotypic subpopulations according to the expression intensity of surface CD25 molecule (IL-2 receptor alpha-chains). Moreover, more recently it has been described that they can be subclassified based on the surface expression of programmed death receptor 1 (PD1). PD-1 is a negative co-stimulatory molecule that plays an important role in the balance and regulation of adaptive immune responses. PD1/PDL1 engagement can down-regulate dramatically T cell activation. AIM: To evaluate Treg cell subtypes in relation with phenotype CD25 and PD1 in H. pylori(+) and H. pylori () biopsies specimens from patients suffering gastritis. METHODS: We analyzed CD4(+) lymphocytes from 16 gastric biopsy specimens of patients with gastritis. H. pylori infection was diagnosed with histology and rapid urease test. Afterwards, it was confirmed by RT PCR assay with SYBR Green I fluorochrome, performed on gastric biopsies using a set of primers for the ureC gene specific for the bacteria. 12 cases of H. pylori(+)-gastritis and 4 of gastritis where H. pylori was not detected were studied. After disintegrating biopsies with a needle, lymphocytes were isolated stirring the remaining tissue in a collagenase-DNase solution (100 U collagenase/mL and 0.1 mg DNase/mL) at 37°C for 2h. The cell suspension was then filtered through mesh and the staining for flow cytometry was performed. The following antibodies were used: anti-PD1-FITC, anti-CD4-PECy5 and anti-CD25-APC. For intracellular labeling with anti-Foxp3-PE, cells were previously permeabilized using Cytofix/ Cytoperm solution. Cells were classified in CD25+ and CD25++ according to the fluorescence of this antibody (cut-off was determined with CD8(+) fluorescence). RESULTS: The percentage of CD4+CD25++Foxp3+ was elevated 5-fold in H. pylori(+) compared to H. pylori(-) samples. The number of CD4+CD25+Foxp3+ was similar in both cases. The subpopulation of CD4+CD25++Foxp3+PD1+ cells in biopsies H. pylori(+) were increased 1.7-fold compared to those found in biopsies H. pylori(-). CONCLUSIONS: H. pylori infection is associated with a marked increase of Treg subpopulation expressing PD1 in gastric mucosa. The use of antibodies anti-PD1 inhibiting only such subset of cells should be investigated as a potential new therapy to reduce gastric inflammation associated with H. pylori infection