Expansion of CD16+ CD56+ NK cells in vericyte® NK cell growth medium
Brohée, Laura ; Bastin, Renaud ; et al
Natural Killer (NK) cells play a key role in host resistance to virus and tumour. These cells are potent killers of virus infected and tumour cells via a direct recognition of the target by activation ... [more ▼]
Natural Killer (NK) cells play a key role in host resistance to virus and tumour. These cells are potent killers of virus infected and tumour cells via a direct recognition of the target by activation receptor such as NKG2D or by inducing Fcγ receptor (FcγRIII, CD16) mediated antibody dependent cellular cytotoxicity (ADCC). Current NK cell-based cancer immunotherapy aims to produce large amounts of functional NK cells, unfortunately most culture media used for NK cell expansion induced the downregulation of CD16 on NK cells. Here, we tested the impact of a new NK cell growth medium (Vericyte® from Medicyte) on CD16 expression. Sorted NK cells and peripheral blood mononuclear cells (PBMC) were cultivated in vericyte® NK cell growth medium and cells issued from these cultures were characterized in term of expansion and phenotype at several time points. After 5 days of culture, an expansion of both NK cells and PBMC was observed and maintained at least until day 20 of culture. In PBMC cultures, we observe only a small preferential NK cell growth since NK cells were around 5-10% at beginning of the culture and this percentage increased to 15% at the end of the culture. However, these cells showed a high proliferative potential when we started the culture with sorted NK cells (the proportion of contaminant cells remain low, under 5%). NK cells expressed CD56 and NKp46 and interestingly after a decreased expression of CD16 on the cell surface at day 3, this receptor was up regulated and most of the cells are CD56bright CD16bright from day 7 to day 12. According FACS FCS/SSC dot plot, NK cells acquired morphology of large activated lymphocytes and some of them expressed activation markers such CD25. Finally, these cells were able to kill efficiently tumour cell line K562. Thus our data show that vericyte® NK cell growth medium allows the expansion of functional CD16+CD56+ NK cells. Cytokine production and ADCC function are under investigation. [less ▲]Detailed reference viewed: 25 (2 ULg)
L'addiction au Soleil, son estocade et la parade des crèmes solaires.
FRANCHIMONT, Claudine ; HERMANNS, Trinh ; PIERARD, Gérald et al
in Revue Médicale de Liège (2013), 68(5-6), 321-325Detailed reference viewed: 11 (1 ULg)
Analytic dermoscopy of superficial basal cell carcinoma.
FRANCHIMONT, Claudine ; ; et al
in Journal of Case Reports in Medicine (2013), 3Detailed reference viewed: 14 (5 ULg)
In vivo evaluation of the skin tensile strength by the suction method: pilot study coping with hysteresis and creep extension.
PIERARD, Gérald ; Pierard, Sébastien ; Delvenne, Philippe et al
in ISRN dermatology (2013), 2013
From an engineering standpoint, both the skin and subcutaneous tissue act as interconnected load-transmitting structures. They are subject to a variety of intrinsic and environmental influences. Changes ... [more ▼]
From an engineering standpoint, both the skin and subcutaneous tissue act as interconnected load-transmitting structures. They are subject to a variety of intrinsic and environmental influences. Changes in the cutaneous viscoelasticity represent an important aspect in a series of skin conditions. The aim of this work was to explore the methodology of biomechanical measurements in order to better appreciate the evolution and severity of some connective tissue diseases. The Cutometer MPA 580 (C+K electronic) was used in the steep and progressive suction procedures. Adapting measurement modalities was explored in order to mitigate any variability in data collection. The repeat steep suction procedure conveniently reveals the creep phenomenon. By contrast, the progressive suction procedure highlights the hysteresis phenomenon. These viscoelastic characteristics are presently described using the 2 and 4 mm probes on normal skin and in scleroderma, acromegaly, corticosteroid-induced dermatoporosis, and Ehlers-Danlos syndrome. The apposition of an additional outer contention on the skin altered differently the manifestations of the creep extension and hysteresis among the tested skin conditions. Any change in the mechanical test procedure affects the data. In clinical and experimental settings, it is mandatory to adhere to a strict and controlled protocol. [less ▲]Detailed reference viewed: 7 (2 ULg)
Dissection of Iliac Artery in a Patient With Autosomal Dominant Polycystic Kidney Disease
Courtois, Audrey ; Nusgens, Betty ; DELVENNE, Philippe et al
in AORTA (2013), 1(2), 123-125Detailed reference viewed: 19 (8 ULg)
Myoferlin is a key regulator of EGFR activity in breast cancer.
Turtoi, Andrei ; Blomme, Arnaud ; Bellahcene, Akeila et al
in Cancer Research (2013)
Myoferlin is a member of the ferlin family of proteins that participate in plasma membrane fusion, repair and endocytosis. While some reports have implicated myoferlin in cancer, the extent of its ... [more ▼]
Myoferlin is a member of the ferlin family of proteins that participate in plasma membrane fusion, repair and endocytosis. While some reports have implicated myoferlin in cancer, the extent of its expression in and contributions to cancer are not well established. In this study, we show that myoferlin is overexpressed in human breast cancers and that it is has a critical role in controlling degradation of the EGFR after its activation and internalization in breast cancer cells. Myoferlin depletion blocked EGF-induced cell migration and epithelial-to-mesenchymal transition. Both effects were induced as a result of impaired degradation of phosphorylated EGFR via dysfunctional plasma membrane caveolae and alteration of caveolin homooligomerization. In parallel, myoferlin depletion reduced tumor development in a chicken chorioallantoic membrane xenograft model of human breast cancer. Considering the therapeutic significance of EGFR targeting, our findings identify myoferlin as an novel candidate function to target for future drug development. [less ▲]Detailed reference viewed: 33 (5 ULg)
Toxic Epidermal Necrolysis and Graft-versus-Host Reaction: Revisiting a Puzzling Similarity.
PIERARD, Gérald ; Hermanns-Lê, Trinh ; PAQUET, Philippe et al
in ISRN Dermatology (2013)Detailed reference viewed: 20 (5 ULg)
Development of anti-HPV lipoplexes for the treatment of cervical cancer
Lechanteur, Anna ; Furst, Tania ; Evrard, Brigitte et al
Poster (2013)Detailed reference viewed: 21 (11 ULg)
The skin landscape in diabetes mellitus. Focus on dermocosmetic management.
PIERARD, Gérald ; ; HERMANNS, Trinh et al
in Clinical, Cosmetic and Investigational Dermatology (2013), 6
BACKGROUND: Some relationships are established between diabetes mellitus (DM) and a series of cutaneous disorders. Specific dermatoses are markers for undiagnosed DM. Other disorders represent supervening ... [more ▼]
BACKGROUND: Some relationships are established between diabetes mellitus (DM) and a series of cutaneous disorders. Specific dermatoses are markers for undiagnosed DM. Other disorders represent supervening complications in an already treated DM patient. OBJECTIVE: To review the information about dermocosmetic care products and their appropriate use in the management and prevention of dermatoses related to DM. METHOD: The peer-reviewed literature and empiric findings are covered. Owing to the limited clinical evidence available for the use of dermocosmetics, a review of the routine practices and common therapies in DM-related dermatoses was conducted. RESULTS: Some DM-related dermatoses (acanthosis nigricans, pigmented purpuric dermatosis) are markers of macrovascular complications. The same disorders and some others (xerosis, Dupuytren's disease) have been found to be more frequently associated with microangiopathy. Other skin diseases (alopecia areata, vitiligo) were found to be markers of autoimmunity, particularly in type 1 DM. Unsurprisingly, using dermocosmetics and appropriate skin care has shown objective improvements of some DM-related dermatoses, such effects improve the quality of life. The most common skin manifestations of DM fall along continuum between "dry skin," xerosis, and acquired ichthyosis, occurring predominately on the shins and feet. Dermocosmetic products improve the feeling of well-being for DM patients. [less ▲]Detailed reference viewed: 15 (1 ULg)
Comment j'explore ... l'atteinte fonctionnelle cutanee d'une sclerodermie.
HERMANNS, Trinh ; Franchimont, Claudine ; PIERARD, Gérald et al
in Revue Médicale de Liège (2013), 68(3), 141-7
Scleroderma refers to distinct clinical presentations sharing in common a sclerotic process most often clinically obvious on the skin. The involvement possibly affects the skin alone in morphea or in ... [more ▼]
Scleroderma refers to distinct clinical presentations sharing in common a sclerotic process most often clinically obvious on the skin. The involvement possibly affects the skin alone in morphea or in combination with internal lesions in systemic sclerosis. Some objective and non-invasive functional assessments are useful for better appreciating the severity and evolution of the disease, as well as to monitor the therapeutic efficacy. In this endeavour, in vivo measurements of the skin mechanical properties are unsurprisingly informative. [less ▲]Detailed reference viewed: 18 (7 ULg)
Ruthenium oligonucleotides, targeting HPV16 E6 oncogene, inhibit the growth of cervical cancer cells under illumination by a mechanism involving p53
Reschner, Anca ; BONTEMS, Sébastien ; et al
in Gene Therapy (2013), 20(4), 435-443Detailed reference viewed: 42 (12 ULg)
Human papillomavirus predicts the outcome following concomitant chemoradiotherapy in patients with head and neck squamous cell carcinomas.
; ; et al
in Oncology Reports (2013), 30(1), 371-6
We investigated the prevalence of human papillomavirus (HPV) in a clinical series of 72 patients with head and neck squamous cell carcinoma (HNSCC) using a retrospective and prospective study design. The ... [more ▼]
We investigated the prevalence of human papillomavirus (HPV) in a clinical series of 72 patients with head and neck squamous cell carcinoma (HNSCC) using a retrospective and prospective study design. The majority of patients were smokers and/or drinkers and were treated with concomitant chemoradiotherapy (CCR). Furthermore, we assessed the impact of HPV positivity on the response to CCR. Paraffin-embedded samples from HNSCC patients (n=72) were evaluated for the presence of HPV DNA using both GP5+/GP6+ consensus PCR and type-specific E6/E7 PCR to detect HPV types 6, 11, 16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 67 and 68. The type-specific E6/E7 PCR demonstrated that 20 out of 69 HNSCC patients (29%) presented with high-risk (HR) HPV types and that 5 of the 69 HNSCC patients (7%) presented with low-risk (LR) HPV types. Using the GP5+/GP6+ PCR, we observed that the rate of response was statistically lower in the HPV+ group (P=0.02). Concerning patient outcomes in terms of recurrence and survival, we observed that the prognosis was poorer for HPV+ patients. We showed for the first time that patients with HPV+ HNSCC present with a worse prognosis after CCR. This observation highlights the need for prospective studies with large numbers of patients and a detailed history of tobacco and alcohol consumption before validating HPV as a marker of prognosis following CCR. [less ▲]Detailed reference viewed: 17 (4 ULg)
Expression of type 2 orexin receptor in human endometrium and its epigenetic silencing in endometrial cancer.
Dehan, Pierre ; ; et al
in Journal of Clinical Endocrinology and Metabolism (2013), 98(4), 1549-57
CONTEXT: Orexins A and B are neuropeptides that bind and activate 2 types of receptors. In addition to direct action in the brain, the orexinergic system has broader implications in peripheral organs, and ... [more ▼]
CONTEXT: Orexins A and B are neuropeptides that bind and activate 2 types of receptors. In addition to direct action in the brain, the orexinergic system has broader implications in peripheral organs, and it has been proposed to have a role in the induction of apoptosis. There are very few data on the endometrium. OBJECTIVE: The expression and epigenetic regulation of type 2 orexin receptor (OX2R) was investigated in the human endometrium as well as in endometrial endometrioid carcinoma (EEC). METHODS: OX2R localization was studied by immunohistochemistry in normal endometrium (n = 24) and in EEC (n = 32). The DNA methylation status of a CpG island located in the first exon of OX2R was analyzed by bisulfite sequencing in normal (n = 18), EEC (n = 34), and 3 endometrial cell lines. On the latter, mRNA expression and Western blotting as well as in vitro induction with orexin were performed. RESULTS: Expression of the OX2R protein was detected in normal endometrial epithelia, whereas it was frequently lacking in EEC. This loss was associated with hypermethylation of OX2R in EEC in comparison with normal endometrium (median CpG methylation percentages of 48.85% and 5.85%, respectively). In cell lines, hypermethylation correlated with weak OX2R expression. Additionally, in vitro treatment of the 3 EEC cell lines with orexins A and B did not result in proliferation change CONCLUSIONS: Altogether our data provide evidence for the epigenetic silencing of OX2R in EEC. The implication of the OX2R loss in tumoral progression remains to be elucidated. [less ▲]Detailed reference viewed: 2 (1 ULg)
Characterization of spontaneous collagen fibrillogenesis in a cell-free and tension-free environment.
PIERARD, Gérald ; Hermanns-Lê, Trinh ; Delvenne, Philippe et al
in Clinical & Experimental Dermatology (2013)
The collagen fibril packing that forms threads and bundles is poorly defined, despite the fact that it is important for distinct aspects of the adventitial and reticular dermis. The present study explored ... [more ▼]
The collagen fibril packing that forms threads and bundles is poorly defined, despite the fact that it is important for distinct aspects of the adventitial and reticular dermis. The present study explored an in vitro fibrillogenesis model using the property of heat polymerization. The process was performed on glass slides with mixtures of collagen I and III, and the material was viewed by scanning electron microscopy. In all instances, collagen I and III formed fibrils with regular sizes. The formation of threads was influenced by the relative proportions of collagen I and III; increasing the relative proportion of collagen I resulted in the formation of threads showing increasing variations in thickness. These findings are in line with the differential presentation and compositions of the different parts of the dermis. The possible interventions of stromal cells and of other macromoleules of the extracellular matrix were not considered in this study. [less ▲]Detailed reference viewed: 11 (1 ULg)
Novel cooperation between CX3CL1 and CCL26 inducing NK cell chemotaxis via CX3CR1: a possible mechanism for NK cell infiltration of the allergic nasal tissue.
EL SHAZLY, Amr ; ; et al
in Clinical & Experimental Allergy : Journal of the British Society for Allergy & Clinical Immunology (2013), 43(3), 322-31
BACKGROUND: Recent data indicated that natural killer (NK) cells and chemokines could play a pivotal role in nasal inflammation. CX3CR1, the only receptor for fractalkine/CX3CL1, is abundantly expressed ... [more ▼]
BACKGROUND: Recent data indicated that natural killer (NK) cells and chemokines could play a pivotal role in nasal inflammation. CX3CR1, the only receptor for fractalkine/CX3CL1, is abundantly expressed by NK cells, and was recently shown to also be a receptor for eotaxin-3/CCL26. However, no reports explored the NK cells-CX3CL1-CCL26 axis via CX3CR1 in allergy. OBJECTIVE: Our goals were first to determine specifically NK cell recruitment pattern in nasal tissue of allergic chronic rhinosinusitis (ACRS) and non-allergic chronic rhinosinusitis (NACRS) patients in comparison with healthy controls, and secondly, to investigate the function of CX3CR1 in NK cell migration. METHODS: Immunohistochemistry, microchemotaxis chambers, flow cytometry and confocal microscopy were used in this study. RESULTS: Herein, we showed that NK cells infiltrated the epithelial layers of nasal tissue only in ACRS patients and not in NACRS patients or controls. NK cells were also more numerous in the stroma of the nasal tissue from ACRS patients compared with NACRS patients or controls. This migration could be mediated by both CX3CL1 and CCL26, as these two chemokines induced NK cell migration. Moreover, both molecules also stimulated cytoskeleton changes and F-actin reorganisation in NK cells. Chemotaxis and cytoskeleton changes were sensitive to genistein, a tyrosine kinase inhibitor. By flow cytometry, we demonstrated that a single antigen nasal provocation challenge increased the expression of CX3CR1 on NK cells in allergic rhinitis (AR) patients. The function of this receptor was associated with a significant augmentation of NK cell chemotaxis against the optimal doses of CX3CL1 and CCL26. CONCLUSIONS AND CLINICAL RELEVANCE: Our results highlight a novel role for CX3CR1 in NK cell migration that may contribute to the NK cell trafficking to the allergic upper airway. This could be mediated largely by CX3CL1 and CCL26 stimulation of the tyrosine kinase pathway. [less ▲]Detailed reference viewed: 23 (3 ULg)
Dendritic cells in Barrett's carcinogenesis: an inadequate environment for antitumor immunity?
Somja, Joan ; Demoulin, Stéphanie ; RONCARATI, Patrick et al
Conference (2013)Detailed reference viewed: 17 (6 ULg)
Evaluation of a new biocompatible poly(N-(morpholino ethyl methacrylate)-based copolymer for the delivery of ruthenium oligonucleotides, targeting HPV16 E6 oncogene
Reschner, Anca ; ; et al
in Journal of Biomedical Nanotechnology (2013), 9
This study investigates the use of a new biocompatible block copolymer poly(2-(dimethylamino)ethyl methacrylate-N-(morpholino)ethyl methacrylate (PDMAEMA-b-PMEMA) for the delivery of a particular ... [more ▼]
This study investigates the use of a new biocompatible block copolymer poly(2-(dimethylamino)ethyl methacrylate-N-(morpholino)ethyl methacrylate (PDMAEMA-b-PMEMA) for the delivery of a particular antisense oligonucleotide targeting E6 gene from human papilloma virus. This antisense oligonucleotide was derivatized with a polyazaaromatic RuII complex which, under visible illumination, is able to produce an irreversible crosslink with the complementary targeted sequence. The purpose of this study is to determine whether by the use of a suitable transfection agent, it is possible to increase the efficiency of the antisense oligonucleotide targeting E6 gene, named Ru-P-4. In a recent study, we showed that Oligofectamine® transfected Ru-P-4 antisense oligonucleotide failed to inhibit efficiently the growth of cervical cancer cell line SiHa, contrarily to the Ru-P-6 antisense oligonucleotide, another sequence also targeting the E6 gene. The ability of PDMAEMA-b-PMEMA to form polyplexes with optimal physicochemical characteristics was investigated first. Then the ability of the PDMAEMA-b-PMEMA/Ru-P-4 antisense oligonucleotide polyplexes to transfect two keratinocyte cell lines (SiHa and HaCat) and the capacity of polyplexes to inhibit HPV16 + cervical cancer cell growth was evaluated. PDMAEMA-b-PMEMA base polyplexes at the optimal molar ratio of polymer nitrogen atoms to DNA phosphates (N/P), were able to deliver Ru-P-4 antisense oligonucleotide and to induce a higher growth inhibition in human cervical cancer SiHa cells, compared to other formulations based on Oligofectamine®. [less ▲]Detailed reference viewed: 86 (24 ULg)
Echocardiographic integrated backscatter for detecting progression and regression of aortic valve calcifications in rats.
; ; et al
in Cardiovascular Ultrasound (2013), 11(1), 4
ABSTRACT: BACKGROUND: Calcification is an independent predictor of mortality in calcific aortic valve disease (CAVD). The aim of this study was to evaluate the use of non-invasive, non-ionizing ... [more ▼]
ABSTRACT: BACKGROUND: Calcification is an independent predictor of mortality in calcific aortic valve disease (CAVD). The aim of this study was to evaluate the use of non-invasive, non-ionizing echocardiographic calibrated integrated backscatter (cIB) for monitoring progression and subsequent regression of aortic valvular calcifications in a rat model of reversible renal failure with CAVD, compared to histology. METHODS: 28 male Wistar rats were prospectively followed during 21 weeks. Group 1 (N=14) was fed with a 0.5% adenine diet for 9 weeks to induce renal failure and CAVD. Group 2 (N=14) received a standard diet. At week 9, six animals of each group were killed. The remaining animals of group 1 (N=8) and group 2 (N=8) were kept on a standard diet for an additional 12 weeks. cIB of the aortic valve was calculated at baseline, 9 and 21 weeks, followed by measurement of the calcified area (Ca Area) on histology. RESULTS: At week 9, cIB values and Ca Area of the aortic valve were significantly increased in the adenine-fed rats compared to baseline and controls. After 12 weeks of adenine diet cessation, cIB values and Ca Area of group 1 decreased compared to week 9, while there was no longer a significant difference compared to age-matched controls of group 2. CONCLUSIONS: cIB is a non-invasive tool allowing quantitative monitoring of CAVD progression and regression in a rat model of reversible renal failure, as validated by comparison with histology. This technique might become useful for assessing CAVD during targeted therapy. [less ▲]Detailed reference viewed: 18 (5 ULg)