Marine seagrass extract of Thalassia testudinum suppresses colorectal tumor growth, motility and angiogenesis by autophagic stress and immunogenic cell death pathways

Marine plants have become an inexhaustible reservoir of new phytopharmaceuticals for cancer treatment. We demonstrate in vitro/in vivo antitumor efficacy of a standardized polyphenol extract from the marine angiosperm Thalassia testudinum (TTE) in colon tumor cell lines (RKO, SW480, and CT26) and a syngeneic allograft murine colorectal cancer model. MTT assays revealed a dose-dependent decrease of cell viability of RKO, CT26, and SW480 cells upon TTE treatment with IC50 values of, respectively, 175, 115, and 60 mu g/mL. Furthermore, TTE significantly prevented basal and bFGF-induced angiogenesis in the chicken chorioallantoic membrane angiogenesis assay. In addition, TTE suppressed bFGF-induced migration of endothelial cells in a wound closure assay. Finally, TTE treatment abrogated CT26 colorectal cancer growth and increased overall organism survival in a syngeneic murine allograft model. Corresponding transcriptome profiling and pathway analysis allowed for the identification of the mechanism of action for the antitumor effects of TTE. In line with our in vitro/in vivo results, TTE treatment triggers ATF4-P53-NF kappa B specific gene expression and autophagy stress pathways. This results in suppression of colon cancer cell growth, cell motility, and angiogenesis pathways in vitro and in addition promotes antitumor immunogenic cell death in vivo

Mixed T helper cell signatures in chronic rhinosinusitis with and without polyps.

In chronic rhinosinusitis (CRS) different phenotypes have been reported based on cytokine profile and inflammatory cell patterns. The aim of this study was to characterize the intracytoplasmatic cytokines of T cells infiltrating the inflamed sinonasal mucosa.Infiltrated T cells and tissue homogenates from sinonasal mucosal samples of 7 healthy subjects, 9 patients with CRS without nasal polyp (CRSsNP), 15 with CRS with nasal polyps (CRSwNP) and 5 cystic fibrosis patients (CF-NP) were analyzed for cytokine expression using flow cytometry and multiplex analysis respectively. Intracytoplasmic cytokinesin T cells were analyzed after stimulation of nasal polyps with Staphylococcus aureus enterotoxin B for 24 hours.The number of T cells per total living cells was significantly higher in patients with CRSwNP vs. CRSsNP and controls. 85% of the CD4(+) T cells showed to be memory T cells. The effector T cells present in all tissues have a predominant Th1 phenotype. Only in CRSwNP, a significant fraction of T cells produced the Th2 cytokines IL-4 and IL-5, while nasal polyps from CF patients were characterized by a higher CD4/CD8 T cell ratio and an increased number of Th17 cells. 24 h stimulation with SEB resulted in a significant induction of CD4(+) T cells producing IL-10 (Tr1 cells).T cell cytokine patterns in healthy and inflamed sinonasal mucosa revealed that Th2 cells (IL-4 and IL-5 producing cells) are significantly increased in CRSwNP mucosal inflammation. Exposure to SEB stimulates Tr1 cells that may contribute to the Th2 bias in CRSwNP

Staphylococcus aureus and Chronic Airway Disease

Staphylococcus aureus (S. aureus) is correlated with the development of persistent severe inflammatory disease of the upper airway including chronic rhinosinusitis with nasal polyps. This inflammation of the upper airways is characterized by a T-helper 2-driven disease: interleukin-5 is significantly increased and local production of immunoglobulin E is observed. S. aureus and its enterotoxins are deregulating the tissue inflammation at different levels: structural cells and the innate and adaptive immune system. Knowing the triggers of the pathomechanisms involved will greatly help us to find new therapeutic approaches to resolve this chronic inflammatory process. Keywords: chronic rhinosinusitis, nasal polyps, Staphylococcus aureus, innate and adaptive immune system, superantigen

Mast cells and nasal mucosa as new targets for leptin action /

Thesis submitted in fulfillment of the requirements for the degree of doctor in medical science

Influence of SEB on T cell cytokine pattern.

<p>Data are expressed in Box-Whisker-plots presenting the results of CRSwNP single suspension after 24 hours without and with 0,5 µg/ml SEB (a). Significance (<i>p</i>) values after <i>Mann-Whitney U</i> test are represented by: *** when p<0,005. Pie charts representing the median result of the different T cells subsets with and without SEB (b).</p

T cell subsets in healthy and diseased nasal mucosa.

<p>Different T cell subsets, namely Th1, Th2, Th17, Th22, Th1/Th17, T cell producing IL-10 or IL-21 and non-producing T cells were analyzed by flow cytometry. Median results for control, CRSsNP, CRSwNP and CF-NP are presented in pie charts. Pie charts presenting the variability in CRSwNP signatures. Scatter plots presenting the cytokine expression by CD4+ T cells without stimulus (a) and with stimulus PMA/Ionomycin (b).</p

Patient description.

<p>Patient description.</p

Cytokine data on tissue homogenates.

<p>Data are expressed as median and interquartile range. P values signs after Mann-Whitney <i>U</i> test means as follow: *: p<0,05 in C versus CRSsNP, C versus CRSwNP or C versus CF-NP; ∇: p<0,05 in CRSsNP versus CRSwNP or CRSsNP versus CF-NP; ◊: p<0,05 in CRSwNP versus CF-NP.</p

Flow cytometricanalyses of T cells in sinonasal mucosa.

<p>Data are expressed in Box-Whisker-plots presenting the results of CD3+ T cells (a), CD4+ T cells (b), CD8+ T cells (c) and the proportion CD45RA+ within the CD3+ T cell population (d). Box-Whisker-plot presenting the results of the percentage of Th2 cells in the different disease groups (e). Percentage of Tc1 cells (IFNγ producing CD8 T cells) in healthy and diseased nasal mucosa are presented as a Box-Whisker-plot (f). Significance (<i>p</i>) values after <i>Mann-Whitney U</i> test are represented by: * when p<0,05, ** when p<0,01 and *** when p<0,005.</p