bluuebunny/crossref_metadata_2025 huggingface dataset:
At Least Three Phosphorylation Events Induced by Pemphigus Vulgaris Sera are Pathogenically Involved in Keratinocyte Acantholysis
Authors: Cirillo N. , Lanza M. , De Rosa A. , Femiano F. , Gombos F. , Lanza A.
Published: January 2017

Intercellular adhesion among keratinocytes is guaranteed by desmosomes. Disruption of desmosomal integrity leads to cell-cell detachment or acantholysis, as it classically occurs in pemphigus vulgaris (PV), an autoimmune blistering disease of skin and mucous membranes. While purified PV IgG seems to trigger intracellular signaling that crucially involves p38 MAPK, keratinocyte acantholysis induced by whole PV serum may recruit a number of additional signals. In this study, the Pro-Q® Diamond Phosphoprotein Assay was used to investigate the overall changes in protein phosphorylation levels in an in vitro model of PV. We showed that keratinocytes exposed to whole PV sera underwent at least three early and transient phosphorylation events. Two bands with apparent molecular masses of ∼35 and ∼45 kDa were found to be phosphorylated within 1 min after incubation with PV sera. A third band of about 80 kDa reached the peak of phosphorylation level after 3 hours. Morphologic evidence of cell shrinkage and acantholysis were late events and did not correlate temporally with kinase activation, suggesting that cytoskeleton reorganization is a downstream phenomenon. Interestingly, pharmacological abrogation of PV-specific protein phosphorylation was able to inhibit the cell-cell detachment, rounding up, and redistribution of Dsg3 in keratinocytes. Thus, at least three phosphorylation events are pathogenically involved in pemphigus acantholysis.
Impact of Different Texture of Polypropylene Mesh on the Inflammatory Response
Authors: Di Vita G. , Patti R. , Sparacello M. , Balistreri C.R. , Candore G. , Caruso C.
Published: January 2017

Over the past decade, hernia surgery has undergone a considerable transformation with the use of prosthetic materials. The most used polypropylene meshes induce a rapid acute inflammatory response followed by chronic foreign body reaction. Many factors influence this response such as density, size, physical characteristics, different texture and porosity of each biomaterial. The aim of this study is to assess whether the implant of monofilament or multifilament meshes, in the inguinal hernioplasty, determine a different inflammatory response. Thirty-two male patients were included in the study and were randomly divided into two groups. In the first group (MO) inguinal hernioplasty was performed using monofilament polypropylene mesh, while in the second one (MU) multifilament prosthesis was used. Peripheral venous blood samples were collected 24 hours before surgery and then 6,24,48 and 168 hours posto-peratively. Modifications in leukocyte count, C-reactive protein (CRP), alpha-1 antitrypsin (α1-AT), interleukin (IL)-1, IL-6, IL-1 ra and IL-10 serum levels were recorded at all sampling times. We present evidence that serum levels of CRP, (α1-AT), leukocytes and cytokines were significantly increased post-operatively in both groups, returning to basal values 168 hours afterwards. In particular, the production of all pro-inflammatory mediators was higher in the MU group, whereas the anti-inflammatory cytokine (IL-10, IL-1ra) production was higher in MO patients. Our results indicate that polypropylene multifilament mesh allows a higher intense acute inflammatory response as compared to monofilament mesh implantation.
Intratumoral vs Systemic Administration of Meta-Tetrahydroxyphenylchlorin for Photodynamic Therapy of Malignant Gliomas: Assessment of Uptake and Spatial Distribution in C6 Rat Glioma Model
Authors: Mannino S. , Molinari A. , Sabatino G. , Ciafrè S.A. , Colone M. , Maira G. , Anile C. , Arancia G. , Mangiola A.
Published: January 2017

Malignant gliomas, with an incidence of 5 cases per 100,000 population per year, represent the most common primary brain tumour. They have an overall survival length of less than 2 years. Many different adjuvant therapies have been developed. Among them, Photodynamic Therapy (PDT), that is based on photochemical reactions between light and tumoral tissue selectively labelled with exogenous photosensitizing agents. Among photosensitizers, m-THPC (Temoporfin), seems to be the most promising one for the treatment of brain tumors, but, unfortunately, it causes problems of high skin photosensitivity. To by-pass this problem, we devised an intratumoral route of administration of this photosensitizer. The aim of this study is to investigate and compare the uptake of m-THPC in brain tumor and normal tissue after systemic and intratumoral administration of the drug. 30 female Wistar rats received m-THPC 12 days after C6 tumor implantation. Temoporfin was administered intratumorally in 24 rats at two different concentrations. 6 rats constituted the control group and received m-THPC by means of an intraperitoneal injection. The brains were extracted at 4 h, 24 h and 96 h after Temoporfin injection. The samples were examined with a confocal laser scanning microscope. All samples showed high fluorescence emission exclusively in the tumour area, without appreciable differences between the samples taken at the different times of sacrifice and the two routes of administration. No fluorescence whatsoever was detected among normal brain tissue surrounding the tumour. The intratumoral route appears to give comparable results to the systemic one, regarding intracellular uptake efficiency and tumour - normal tissue ratio, with the advantage of a much shorter time needed to reach optimal intratumoural concentration - that is just four hours from m-THPC injection.
Dilated Cardiomyopathy Due to Thrombotic Microangiopathy as the only Manifestation of Antiphospholipid Syndrome: A Case Report
Authors: Vaccaro F. , Caccavo D. , Roumpedaki E. , De Vincentis G. , Di Gioia C.R.T. , Gallo P. , Palange P.
Published: January 2017

Dilated cardiomyopathy due to thrombotic microangiopathy has been rarely reported as a clinical manifestation of antiphopspholipid syndrome (APS). We describe the case of a 39-year-old woman affected by systemic lupus erythematosus (SLE) and positive antiphospholipid antibodies (aPL) who presented with orthopnea and peripheral oedema. Diagnosis of dilated cardiomyopathy due to myocardial thrombotic microangiopathy was made and treatment with anticoagulants prevented the worsening of the clinical condition. Interestingly, at variance with other cases, our patient showed no extracardiac signs of APS. The review of the current literature has confirmed that dilated cardiomyopathy due to thrombotic microangiopathy is a rare manifestation of APS.
Hypoxemia with High Alveolar-Arterial Oxygen Gradient but no Lung Involvement in a Patient with Churg-Strauss Syndrome: Case Report
Authors: Utsugi M. , Ishizuka T. , Hisada T. , Sato K. , Ishizuka T. , Dobashi K. , Mori M.
Published: January 2017

Churg-Strauss syndrome (CSS) is characterized by asthma and/or a history of allergy, eosinophilia and an often life-threatening systemic necrotizing vasculitis. We describe a patient with CSS and hypoxemia with a high alveolar-arterial oxygen gradient (AaDO2), but no pulmonary parenchymal involvement. The patient also had a low diffusion capacity with normal lung volume and a high level of serum thrombomodulin, a marker of endothelial cell injury. Treatment for CSS, such as corticosteroid, improved both hypoxemia and AaDO2 consistent with amelioration of diffusion capacity and serum thrombomodulin level, suggesting that this pathosis involves microangiopathy with endothelial cell damage induced by vasculitis in pulmonary blood vessels.
Clinical Evaluation and Treatment of Acute Asthma Exacerbations in Children
Authors: Indinnimeo L. , Bertuola F. , Cutrera R. , De Benedictis F.M. , Di Pietro P. , Duse M. , Gianiorio P. , Indirli G. , La Grutta S. , La Rosa M. , Longhi R. , Sopo S. Miceli , Miglioranzi P. , Del Giudice M. Miraglia , Monaco F. , Radzik D. , Renna S. , Snijders D. , Zampogna S. , Barbato A.
Published: January 2017

This update on treatment of asthma exacerbations in children is the result of an Italian Pediatric Society Task-force, made up of a panel of experts working in 2007–2008. The aim is to give clear indications on the use of the drugs most employed in children, grading the quality of evidence and the strength of recommendations. Suggestions on their limits due to unlicensed and off-label use are reported. The level of evidence and the strength of recommendations for different therapeutic approaches demonstrate that frequently the use of drugs in children is extrapolated from the experience in adults and that more studies are required to endorse the correct use of different drugs in asthmatic children.
Suppression of Map Kinases Inhibits Microglial Activation and Attenuates Neuronal Cell Death Induced by α-Synuclein Protofibrils
Authors: Wilms H. , Rosenstiel P. , Romero-Ramos M. , Arlt A. , Schäfer H. , Seegert D. , Kahle P.J. , Odoy S. , Claasen J.H. , Holzknecht C. , Brandenburg L.O. , Deuschl G. , Schreiber S. , Kirik D. , Lucius R.
Published: January 2017

α-Synuclein (α-Syn) accounts, as a major component of Lewy bodies (LB), for the filamentous deposits in many cases of neurodegenerative diseases. Yet, little is known about the molecular mechanisms of neuronal loss in these diseases. The correlation between α-Syn oligomerization/aggregation and pathologies raises the key question of which molecular form of α-Syn (i.e. monomeric α-Syn, protofibrils or mature fibrils) represents the damage-inducing culprit in the scenario of synucleinopathies. We show that human α-Syn protofibrils (PFs) are potent activators of parallel proinflammatory signalling pathways (p38 and ERK1/2 MAP kinases and NF-κB) in microglial cells in vitro. Furthermore, stereotactic injection of α-Syn PFs into the substantia nigra of adult rats leads to a profound activation of microglia and adjacent neuronal cell loss, which can be attenuated by the MAP kinase inhibitor semapimod. We propose that the neurodegenerative process of α-synucleinopathies involves microglial activation through α-Syn released or extruded from cells with pathogenic α-Syn metabolism. Compounds that inhibit the MAPK/NF-κB pathways might be a promising pharmacological strategy for the treatment of the inflammatory component of synucleinopathies including PD.
Effect of PD98059, a Selective MAPK3/MAPK1 Inhibitor, on Acute Lung Injury in Mice
Authors: Di Paola R. , Crisafulli C. , Mazzon E. , Genovese T. , Paterniti I. , Bramanti P. , Cuzzocrea S.
Published: January 2017

The aim of the present study is to evaluate the contribution of mitogen-activated protein kinase 1–3 (MAPK3/MAPK1) in a model of acute lung inflammation in mice. Injection of carrageenan into the pleural cavity of mice elicited an acute inflammatory response characterized by: accumulation of fluid containing a large number of neutrophils (PMNs) in the pleural cavity, infiltration of PMNs in lung tissues and subsequent adhesion molecule expression (I-CAM and P-selectin), lipid peroxidation, and increased production of tumour necrosis factor-α, (TNF-α) and interleukin-1β (IL-1β). Furthermore, carrageenan induced lung apoptosis (Bax and Bcl-2 expression) as well as nitrotyrosine formation, NF-κB activation, and pJNK expression, as determined by immunohistochemical analysis of lung tissues and the degree of lung inflammation and tissue injury (histological score). Administration of PD98059, an inhibitor of MAPK3/MAPK1 (10 mg/kg) 1 h after carrageenan caused a reduction in all the parameters of inflammation measured. Thus, based on these findings we propose that inhibitors of the MAPK3/MAPK1 signaling pathways, such as PD98059, may be useful in the treatment of various inflammatory diseases.
Moraxella Catarrhalis-Specific Th1 Cells in Bal Fluids of Chronic Obstructive Pulmonary Disease Patients
Authors: Amedei A. , Della Bella C. , Niccolai E. , Stanflin N. , Benagiano M. , Duranti R. , Del Prete G. , Murphy T.F. , D'Elios M.M.
Published: January 2017

In chronic obstructive pulmonary disease (COPD) patients' airway mucosa is infiltrated by macrophages and T lymphocytes, potentially reactive to pathogens. We studied the antigen-specificity and the effector functions of in vivo activated T lymphocytes isolated from BAL (Bronchoalveolar lavage) of 5 Moraxella catarrhalis (Mc)-infected and 5 Mc-non-infected COPD patients. Mc-specific T cells were detected only in BAL or peripheral blood of Moraxella catarrhalis-infected patients. The majority of BAL Mc-specific T cells expressed the T helper type 1 (Th1) cytokine profile with high cytotoxic and pro-apoptotic activity. Upon antigen stimulation, all Mc-specific T clones were able to help the immunoglobulin production by autologous B cells and the MMP (Matrix MetalloProteinase)-12 activity by monocytes. Our results suggest a role for Th1-driven response to Moraxella catarrhalis in the genesis of COPD.
Endothelial Progenitor Cells are Mobilized after Major Laparotomic Surgery in Patients with Cancer
Authors: Schillaci G. , Rondelli F. , Pirro M. , Bagaglia F. , Pucci G. , Noya G. , Mannarino E.
Published: January 2017

The progression of cancer is largely dependent on neoangiogenesis. Circulating endothelial progenitor cells (EPC) have the ability to form complete vascular structures in vitro and play a crucial role in tumor vasculogenesis. Emerging evidence suggests that surgical injury may induce the mobilization of EPC in animal models, and this might have a negative effect on the prognosis of cancer patients. We studied 20 patients (10 men, 65±13 years) undergoing laparotomy for surgical treatment of various forms of abdominal cancer, and 20 age- and sex-matched healthy control subjects. The number of circulating EPC, defined as CD34+/KDR+ cells identified among mononuclear cells isolated from peripheral venous blood, was determined preoperatively and at days 1 and 2 after surgery. Surgery induced a significant increase in circulating EPC levels at day 1 (from 278/mL, interquartile range 171–334, to 558/mL, interquartile range 423–841, p<0.001) and day 2 (709/mL, interquartile range 355–834, p<0.001) compared with baseline values. EPC levels did not change in control subjects. Seven subjects who underwent laparotomic surgery for non-neoplastic disease also showed an increase in EPC levels after surgery (p=0.009 and p=0.028 at day 1 and day 2, respectively). We conclude that patients undergoing elective laparotomic surgery for cancer demonstrate an increase in EPC post-operatively. The potential adverse effects of surgical stress-induced EPC mobilization on tumor and metastasis growth in cancer patients need to be addressed in future studies.
Natural Killer Cell Activity Decreases in Workers Occupationally Exposed to Extremely Low Frequency Magnetic Fields Exceeding 1 μt
Authors: Gobba F. , Bargellini A. , Bravo G. , Scaringi M. , Cauteruccio L. , Borella P.
Published: January 2017

In a preliminary study a reduction in Natural Killer (NK) cell activity in peripheral blood lymphocytes (PBL) was observed in a group of workers exposed to levels of Extremely Low Frequency-Magnetic Fields (ELF-MF) exceeding 1 μT. This study was performed to confirm the results. In 121 workers engaged in various occupational activities, individual ELF-MF exposure was monitored for 2 work shifts. Exposure levels were calculated as Time-Weighted Average (TWA). Subjects were classified as Low exposure (TWA ≤ 0.2 μT), Medium exposure (TWA 0.21–0.99 μT), or Higher exposure (TWA ≥1 μT). In higher exposure workers NK activity proved significantly reduced compared to low exposure, (p<0.01). In medium exposure a reduction was also observed, but the difference was not significant. Multivariate analysis also confirmed the relation between exposure and NK activity. It has been suggested that ELF might affect tumour progression by inducing changes in the immune system: due to the role played by NK activity in host defence against cancer, the interference with the NK cell activity observed in this study is in agreement with this hypothesis. Furthermore, an increased risk for some neurodegenerative disorders has been reported in some epidemiological studies in ELF-MF-exposed workers: changes in NK function were also described in these diseases. Our results, showing the effect on NK activity of exposure exceeding 1 μT, suggest a possible mechanism for ELF-MF effects. This could open new horizons regarding the adverse long-term effects of these fields.
Vascular Endothelial Growth Factor and E-Nitric Oxide Synthase-Mediated Regenerative Response Occurring upon Autologous and Heterologous Bone Grafts
Authors: Tetè S. , Zara S. , Vinci R. , Zizzari V. , Di Tore U. , Mastrangelo F. , Cataldi A. , Gherlone E.
Published: January 2017

Bone regeneration procedures allow oral rehabilitation with dental implants also in edentulous ridges with severe bone atrophy. The integration of grafted materials with the host tissue can initiate regenerative, inflammatory and apoptotic response. Since molecular mechanisms exist at the basis of such response, the aim of this work is to investigate, by immunohistochemical analyses, the expression of proteins involved in the graft integration process, in parallel to clinical and histological modifications, occurring on sites treated with extraoral autologous bone graft deriving from the parietal region of the calvaria (eAB), intraoral autologous bone graft deriving from mandibular ramus (iAB) and heterologous bone graft from swine (hB) in human patients. In our study, the immunohistochemical expression of BSP, VEGF, eNOS in eAB samples was significantly higher (p< 0.05) compared to values recorded in iAB and hB samples. The inflammatory response, investigated by iNOS expression, was found lower in all autologous samples (eAB and iAB) compared to hB, at statistically significant values. Moreover, the expression of the pro-apoptotic molecule, Bax, resulted significantly lower (p< 0.05) in eAB than in iAB and hB samples. These values, together with the low number of apoptotic cells detected in autologous samples, suggest a good regenerative response when extraoral autologous bone graft is used in comparison to the response from the other grafts, and also suggest the use of calvaria graft as a predictable therapeutic procedure for repairing severe bone defects in oral and maxillofacial surgery, not only by clinical and biomechanical criteria, but also from a biomolecular aspect.
Bevacizumab-Related Osteneocrosis of the Jaw
Authors: Serra E. , Paolantonio M. , Spoto G. , Mastrangelo F. , Tetè S. , Dolci M.
Published: January 2017

We describe a case of jaw bone necrosis after a lung adenocarcinoma bone metastasis, treated the first time in 2004 by means of pneumonectomy and lymph node ablation. One week after a dental extraction, the patient experienced pain in the mandibular region, in conjunction with alveolar bone exposure. Treatment with amoxicillin and clavulanate every 12 hours for 15 days and 0.2% chlorhexidine rinses was administered and there was a remission of infective complications, but not the closure of the exposed alveolar bone. Only at this time did the patient refer that he was treated with bevacizumab therapy immediately after the extraction. A preventive dental assessment of patients scheduled for bevacizumab therapy should be useful as for the zoledronic acid therapy. Dental surgery procedures for patients during bevacizumab therapy should be carefully evaluated and considered as the last choice, to reduce all possible risks and prevent complications.
Combination Treatment with Etanercept and an Intensive SPA Rehabilitation Program in Active Ankylosing Spondylitis
Authors: Colina M. , Ciancio G. , Garavini R. , Conti M. , Trotta F. , Govoni M.
Published: January 2017

The aim of this study is to determine the effects of a combination treatment with etanercept and spa rehabilitation versus etanercept alone on function, disability and quality of life in a group of patients with active ankylosing spondylitis (AS). Sixty patients with AS underwent etanercept as suggested by ASAS/EULAR recommendations. As the clinical and laboratory conditions improved, 30 patients accepted the proposal of coupling the medical therapy with a 7-day rehabilitation program in a thermal baths centre; the remaining 30 subjects continued to take the biologic agent alone. The comparisons between the 2 groups were made after 3 and 6 months. The primary outcome was an improvement in BASFI. The secondary outcome was an improvement in the visual analogic scale of EuroQol (EQ-5Dvas). After 6 months a statistically significant improvement in BASFI (p < 0.05) and EQ-5DVAS (p < 0.05) scores was observed in both groups. The mean change in EQ-5DVAS value showed a statistically significant difference in favour of the combination therapy group versus the monotherapy group (22 vs 32, p < 0.05). A therapeutic regimen combining etanercept with an intensive rehabilitation program contributes to disability reduction and ameliorates quality of life for AS patients.
Long-Term Tolerability of Etoricoxib in Patients with Previous Reactions to Non-Steroidal Anti-Inflammatory Drugs
Authors: Di Leo E. , Aloia A.M. , Nettis E. , Cardinale F. , Foti C. , Distaso M. , Ferrannini A. , Vacca A.
Published: January 2017

Non-steroidal anti-inflammatory drugs frequently cause adverse reactions. This retrospective study was based on analysis of the data obtained from interviews conducted with 173 patients, who underwent and tolerated a challenge test with etoricoxib (a selective cyclooxigenase 2 enzyme inhibitor). Only one of 82 patients who were treated with etoricoxib reported reactions. We can conclude that etoricoxib shows a high long-term tolerability in patients with non-steroidal anti-inflammatory drug hypersensitivity.
Life Expectancy of Women with Lupus Nephritis Now Approaches That of the General Population
Authors: Stratta P. , Mesiano P. , Campo A. , Grill A. , Ferrero S. , Santi S. , Besso L. , Mazzucco G. , Rosso S. , Spitale A. , Fop F. , Ciccone G.
Published: January 2017

Immunosuppressive treatment has changed the prognosis of Lupus nephritis over time, but improvement in prognosis is difficult to analyze in different historical periods, and should be better demonstrated in comparison with life expectancy of sex-and age-matched people. Long-term patient and renal survival of 90 patients diagnosed with Lupus nephritis at our center from 1968 to 2001 with a follow-up time of 14±8 years was retrospectively evaluated. Patient and kidney survival significantly increased over time. Multivariate analyses show that risks of patient and renal death decreased by 8% at each year of follow-up, and increased by more than 5 time in patients aged > 30 years at diagnosis. As only 14 patients were men, relative survival as compared to that of the sex- and age-matched general population of the Piedmont Region was calculated for the 76 women. Improvement in the survival of the cohort of women was seen at any time of follow-up: in particular, it was sharply lower in the first period (relative survival at 5,10 and 15 years = 0.784, 0.665, and 0.620, respectively) and increased in the second (relative survival at 5,10 and 15 years = 0.939, 0.921, and 0.850, respectively) nearly approaching that expected for the general population, i.e. 0.993, 0.983 and 0.967, respectively. Taken together, our data allow us to draw the conclusion that life expectancy in women with Lupus nephritis has improved over time, paralleling an improved awareness of the disease and a significant increase in steroid pulse therapy as induction/remission phase. Improvement in survival is for the first time demonstrated to cover the gap with life expectancy of the general population for women with Lupus nephritis.
Mixed Panniculitis Responding to Cyclosporin-A with a 12-Month Follow-up: A Case Report
Authors: Migliore A. , Bizzi E. , Santacroce C.M. , Tarquini E. , Massafra U. , Vacca F. , Martin L.S. Martin
Published: January 2017

Panniculitides represent a heterogeneous group of inflammatory diseases involving subcutaneous fat. Subcutaneous fat is normally organized into adipose cells, adipocytes, and septa of connective tissue. The inflammation involving such tissues can be more represented in septa (septal panniculitis) or in lobules (lobular panniculitis) or be equally distributed in both (mixed panniculitis). A bioptical study is necessary in order to discern between different forms. Vascular involvement is also different in such diseases, as it can interest arteries, or veins, or both. Different grades of fat necrosis can also be observed, such as adipocytes without nuclei, lipophagic necrosis, liquefactive fat necrosis, microcystic fat necrosis, ischaemic fat necrosis. Panniculitis can be idiopathic or secondary to other diseases such as systemic sclerosis, rheumatoid arthritis, systemic erithematous lupus and many others. Therapies usually vary on the single patient but the general orientation leads to the use of immunosuppressive drugs such as thalidomide, corticosteroids, cyclosporin-A, hydroxychloroquine and cyclophosphamide. We report a case of a 19-year-old female affected by primary mixed panniculitis, associated with fever and deep asthenia, that resolved in a few weeks and was maintained with oral cyclosporin-A.
Detailed Characterisation of CB2 Receptor Protein Expression in Peripheral Blood Immune Cells from Healthy Human Volunteers Using Flow Cytometry
Authors: Graham E.S. , Angel C.E. , Schwarcz L.E. , Dunbar P.R. , Glass M.
Published: January 2017

It is commonly accepted from gene expression studies that the CB2 receptor is expressed by most cell types of the rodent and human immune system. However, the exact identity of cells expressing CB2 receptor protein in human blood or the abundance of receptors expressed by each immune subset is not well characterised. We conducted a detailed analysis of CB2 protein levels expressed by blood-derived immune cells from healthy human donors. Flow-cytometry was conducted using 4 commercially available anti-CB2 polyclonal antibodies in conjunction with a selection of immune cell specific markers. Across multiple healthy subjects we observed that NK cells, B-lymphocytes and monocytes expressed a higher level of CB2 receptor than CD4+ or CD8+ T-lymphocytes. Neutrophils also expressed a low level of CB2 receptor. NK cells had the greatest variation in CB2 expression levels, whereas for each of the other cell types CB2 levels were relatively similar between subjects. In contrast to other methods, the high sensitivity of flow-cytometry revealed that CB2 receptors are present on resting T-lymphocytes at low abundance in some healthy subjects. These data provide the first detailed analysis of CB2 protein levels in blood leukocyte subsets from healthy donors and identifies the cell types which could be targeted with CB2-mimetic drugs in humans.
Increased Tumor-Specific CD8+ T Cell Induction by Dendritic Cells Matured with a Clinical Grade TLR-Agonist in Combination with IFN-γ
Authors: Vanderlocht J. , Van Elssen C.H.M.J. , Senden-Gijsbers B.L.M.G. , Meek B. , Cloosen S. , Libon C. , Bos G.M.J. , Germeraad W.T.V.
Published: January 2017

The limited response rate of cancer patients treated with dendritic cell (DC)-based vaccines indicates that vast improvements remain necessary. In many murine tumour models it has been demonstrated that the use of innate triggers (e.g. TLR triggers) in the maturation of DC results in higher efficacy. However, as few of these innate triggers are generated clinical grade, there remains a great necessity to fill the gap between fundamental mouse studies and a clinical trial in humans. In the present study we used a TLR2/4-agonist (FMKp which is available clinical grade) in combination with IFN-γ (FI-cocktail) in the maturation of elutriated monocyte-derived DC and compared it with the most used DC in current clinical trials (TNF-α/PGE-2, i.e. TP-cocktail). In addition to the assessment of CD4+ T cell polarizing capacity, we compared the quantity and intrinsic quality of induced CD8+ T cells of 2 different DC maturation protocols with all cells from the same donor. Besides differences in the cytokine profile, which could be coupled to increased Th1 and Th17 polarization, we demonstrate in this study that FMKp/IFN-γ matured DC are twice as effective in inducing cytotoxic T cells against known tumor antigens. Both DCs induced phenotypically equivalent effector memory CD8+ T cells that did not show a significant difference in their intrinsic capacity to kill tumor cells. These findings point to the therapeutic applicability of FI-DC as superior inducers of functional antigen-specific T cells. Their increased chemokine secretion is suggestive of a mechanism by which these DC may compensate for the limited migration observed for all ex vivo cultured DC when applied in patients.
In Vitro Inhibition of Lipopolysaccharide and Mycobacterium Bovis Bacillus Calmette Guérin-Induced Inflammatory Cytokines and in Vivo Protection from D-Galactosamine/Lps -Mediated Liver Injury by the Medicinal Plant Sclerocarya Birrea
Authors: Fotio A.L. , Olleros M.L. , Vesin D. , Tauzin S. , Bisig R. , Dimo T. , Nguelefack T.B. , Dongo E. , Kamtchouing P. , Garcia I.
Published: January 2017

Sclerocarya birrea is a medicinal plant used for the treatment of inflammatory- and bacterial-related diseases. The present study investigated in vitro and in vivo the effects of the stem bark methanol extract of S. birrea. Nitrite, TNF, IL-1β, IL-6 and IL-12p40 production by bone marrow-derived macrophages (BMDM) pre-incubated with or without S. birrea, and stimulated with Lipopolysaccharide (LPS) or infected with live Mycobacterium bovis Bacillus Calmette Guérin (BCG) was evaluated. S. birrea extract inhibited, in a concentration-dependent manner, nitrite, TNF, IL-1β, IL-6 and IL-12p40 production by BMDM stimulated with LPS or infected with live BCG. The iNOS expression was reduced by S. birrea after stimulation of BMDM with LPS. In addition, S. birrea inhibited the nuclear factor κB (NF-κB) activation by both LPS and BCG. The effects of the plant extract were also evaluated in an in vivo model of liver injury induced by D-galactosamine/LPS (D-GalN/LPS) administration in mice. S. birrea limited D-GalN/LPS-liver injury as assessed by a reduction in transaminases and TNF, IL-1β, IL-6 serum levels, and translocation of NF-κB to the nucleus. Taken together, our data indicate that stem bark methanol extract of S. birrea possesses anti-inflammatory properties by inhibiting NF-κB activation and cytokine release induced by inflammatory or infectious stimuli.
Clinicopathological and Therapeutic Significance of CXCL12 Expression in Lung Cancer
Authors: Imai H. , Sunaga N. , Shimizu Y. , Kakegawa S. , Shimizu K. , Sano T. , Ishizuka T. , Oyama T. , Saito R. , Minna J.D. , Mori M.
Published: January 2017

Interactions between CXCL12 and its receptors CXCR4 or CXCR7 are involved in tumor growth and metastasis in various types of human cancer. However, CXCL12 expression and its role in lung cancer are not fully elucidated. Here we examined the expression of CXCL12 in 54 lung cancer cell lines consisting of 23 small cell lung cancers (SCLCs) and 31 non-small cell lung cancers (NSCLCs). CXCL12 was overexpressed in lung cancer cell lines compared to non-malignant human bronchial epithelial cell lines (N = 6). CXCL12 expression was positively but weakly correlated with the expression of CXCR4 or CXCR7. We also examined CXCL12 expression in 89 NSCLC specimens and found that CXCL12 expression was significantly higher in tumor specimens from female patients, non-smokers and adenocarcinoma patients. Small interfering RNAs targeting CXCL12 inhibited cellular proliferation, colony formation and migration of CXCL12-overexpressing lung cancer cells; however, this inhibition did not occur in lung cancer cells that lacked CXCL12. Furthermore, the anti-CXCL12 neutralizing antibody mediated inhibitory effects in three lung cancer cell lines that overexpressed CXCL12, but not in two CXCL12 non-expressing lung cancer cell lines nor two non-malignant bronchial epithelial cell lines. The present study demonstrates that: CXCL12 is concomitantly overexpressed with CXCR4 or CXCR7 in lung cancers; CXCL12 is highly expressed in NSCLCs from females, non-smokers and adenocarcinoma patients; and disruption of CXCL12 inhibits the growth and migration of lung cancer cells. Our findings indicate that CXCL12 is required for tumor growth and provide a rationale for the anti-CXCL12 treatment strategy in lung cancer.
Isolation and Characterization of Circulating Tissue Transglutaminase-Specific T Cells in Coeliac Disease
Authors: Ciccocioppo R. , Finamore A. , Mengheri E. , Millimaggi D. , Esslinger B. , Dieterich W. , Papola F. , Colangeli S. , Tombolino V. , Schuppan D. , Corazza G.R.
Published: January 2017

Tissue transglutaminase (TG2) was identified as the humoral autoantigen in coeliac disease, but whether it can also serve as T cell autoantigen is still unknown. We aimed, therefore, to firstly explore the presence of TG2-specific T cells in peripheral blood of ten adult patients (four active, i.e. carrying both serological and histological features of the disease; four treated, i.e. with proven mucosal recovery and disappearance of specific antibodies after an adequate period of gluten free diet; and two potential coeliacs, i.e. carrying the serological stigmata of the disease, but not the intestinal lesions), and four healthy controls (two carrying the HLA-DQ2 haplotype of susceptibility to the disease), and secondly to carry out a detailed in vitro characterization of the isolated antigen-specific T cells. T cell lines were first established by means of weekly stimulation with human recombinant TG2 followed by generation of T cell clones through distribution of T cells on plates at one cell/well limiting dilution and further rounds of stimulation. Antigen specificity and HLA-DQ2 restriction were both assessed by evaluating the proliferative response to TG2 in the absence and presence of human sera blocking HLA-DQ2 molecules, after exclusion of impurities in the antigen preparation. Immune phenotyping of T cell clones was performed by flow cytometry, and the expression of IL-1β, IL-4, IL-6, IL-10, IL-12, TGF-β, IFN-γ and TNF-α was determined by ELISA assay on the supernatants of these clones. A total of 91 T cell clones were isolated from the three HLA-DQ2-positive, active patients, but none from the other patients and controls. The immune phenotyping showed that the majority of them (85.7%) were CD3/CD4+ and only a small percentage (14.3%) were CD3/CD8+, all carried the TCR αβ, and had a memory phenotype. The cytokine profile showed high levels of IFN-γ and IL-6 that, together with the absence of IL-4, placed these T cell clones in the T helper type 1-like category. Further in vitro analysis was carried out on 32/91 CD4+ clones and showed a specific and dose-dependent proliferative response towards TG2 and an HLA-DQ2 restriction. Finally, when incubating duodenal mucosal specimens of treated patients with the supernatant of TG2-specific T cell clones, characteristic disease lesions were found, indicating a role for TG2-specific cellular immune response in the pathogenesis of coeliac disease.
Soluble CD30: A Biomarker for Evaluating the Clinical Risk versus Benefit of IFNβ1A Treatment in Multiple Sclerosis Patients
Authors: Contasta I. , Totaro R. , Berghella A-M. , Pellegrini P. , Del Beato T. , Carolei A. , Adorno D.
Published: January 2017

Aberrant redox regulation occurs in immune and neurological pathologies, hence targeting the pathways involved in the regulation of the redox system could provide further insights into these diseases and open up new avenues for therapy. Soluble (s) CD30 is of key clinical importance in this respect, as its levels reflect the functionality of the CD30 receptor (CD30R), the specific lymphocyte receptor for thiol disulfide/oxidoreductase thioredoxin 1 (Trx1) which is known to regulate important immune and neurological processes. Increased levels of sCD30 appear to be a common element of oxidative stress, immunological alterations and neurological deficit, therefore these increases could be used as a clinical biomarker and target for therapy. We targeted sCD30 in our study of dendritic cell (DC) regulation of the T helper (Th) cell network in multiple sclerosis (MS) patients, as abnormalities in T regulatory (Treg)/Th1/Th17 pathways contribute to the pathogenesis of this immunological/neurological disease. DC profiles in Treg/Th1/Th2/Th17-types of cytokine production in culture supernatants were used as they determine the type of Th differentiation. Our results show that sCD30 levels increase significantly in MS patients, reflecting the disruption in the regulation of the Treg/Th1/Th17 cell network. A fall in the level of soluble CD30, induced by IFNβ1a therapy, opposed the increase of neurological deficit through increasing IL10 and TGFβ levels, thus re-establishing network homeostasis but only when this was accompanied by an increase in IL12p70 levels. Since IL12p70 cytokine production is regulated by Trx1, our results indicate that redox system alterations may be the cause of IFNβ1a therapeutic inefficacy. We conclude that an increase in the level of IL10, TGFβ and IL12p70 and a fall in the level of sCD30 represent a means of evaluating the clinical risk/benefit of IFNβ1a treatment.
The Osteopontin Gene +1239A/C Single Nucleotide Polymorphism is Associated with Type 1 Diabetes Mellitus in the Italian Population
Authors: Chiocchetti A. , Orilieri E. , Cappellano G. , Barizzone N. , D'Alfonso S. , D'Annunzio G. , Lorini R. , Ravazzolo R. , Cadario F. , Martinetti M. , Calcaterra V. , Cerutti F. , Bruno G. , Larizza D. , Dianzani U.
Published: January 2017

Secreted phosphoprotein 1, also known as Osteopontin (Opn), is a proinflammatory cytokine involved in the TH1 response and is highly expressed in the islets and pancreatic lymph nodes of non-obese diabetic mice before the onset of diabetes. In humans, typing of the +1239A/C single nucleotide polymorphism (SNP) in the 3'UTR of the Opn gene (SPP1) showed that +1239C carriers displayed higher Opn serum levels than +1239A homozygotes and a higher risk of developing autoimmune/lymphoproliferative syndrome, multiple sclerosis, and systemic lupus erythematosus. The aim of this work is to evaluate whether +1239A/C is also associated with type 1 diabetes mellitus (T1DM). We typed +1239A/C in an initial cohort of 184 T1DM patients and 361 controls, and confirmed our data in a second cohort of 513 patients and 857 controls. In both cohorts, +1239C carriers displayed a significantly higher risk of T1DM than +1239A homozygotes (combined cohorts: OR=1.63, 95%CI: 1.34–1.97). Clinical analysis did not detect any differences between patients carrying or not +1239C in terms of gender distribution and age at T1DM diagnosis. These data suggest that SPP1 variants marked by +1239C are associated with T1DM development in the Italian population. The predisposing effect may depend on its effect on Opn levels.
Undifferentiated Connective Tissue Disease - An Unsolved Problem: Revision of Literature and Case Studies
Authors: Conti V. , Esposito A. , Cagliuso M. , Fantauzzi A. , Pastori D. , Mezzaroma I. , Aiuti F.
Published: January 2017

In clinical practice, patients with a range of signs and symptoms suggestive of connective tissue disease, but who do not fulfil the classification criteria for a defined disease are often found. This condition is defined as “Undifferentiated Connective Tissue Disease” (UCTD). Most of the authors consider UCTD as a distinct clinical entity, generally stable during follow-up. Despite this, no mutual agreement regarding criteria for its diagnosis has been reached. The clinical, serological, therapeutical and evolutional patterns of 41 patients initially diagnosed as having early UCTD during a 3-year follow-up are described in this study. At the end of the observational period, 21% of the enrolled patients, followed throughout the follow-up, demonstrated clinical evolution to a defined connective tissue disease (CTD), whereas 52% of the observed subjects maintained an undifferentiated profile with variable clinical findings and presenting a generally stable disease over time. The remaining patients showed clinical improvement or complete regression of the symptoms associated with normalization of the inflammatory indexes. The role of therapy in these different clinical courses is discussed.
A Split-Mouth Study on Microbiological Profile in Clinical Healthy Teeth and Implants Related to Key Inflammatory Mediators
Authors: Schierano G. , Pejrone G. , Roana J. , Scalas D. , Allizond V. , Martinasso G. , Pagano M. , Canuto R.A. , Cuffini A.M.
Published: January 2017

This split-mouth study investigated the correlation of the qualitative and quantitative bacterial composition in dental plaque around clinically healthy periodontal and peri-implant subgingival sites with the levels of selected pro- and anti-inflammatory cytokines and the inflammatory infiltrate in the soft tissue surrounding a healthy dental implant and natural tooth in the same patient. Nine patients, all in good health and non-smokers, were studied. All of the patients were highly motivated in terms of oral hygiene and had healthy natural teeth and at least one healthy implant. After three sessions of professional oral care, clinical parameters were recorded. A sample of subgingival plaque was harvested with a sterile curette from the buccal side of the selected implants and teeth. The plaque samples were cultured to quantify the total microbiota and the number of obligate and facultative bacterial strains. Simultaneously, from the lingual/palatal aspect of the same implants and teeth the keratinized periodontal and peri-implant soft tissues were biopsied for cytokine expression and histomorphometric analysis. The tissue biopsies were halved: the real-time reverse transcriptase-polymerase chain reaction (PCR) was performed to detect active TNF-α, IL-1β, IL-8, and TGF-β2 and distribution, composition, quantification of inflammation were assessed in parallel. The patients harbored no periodontopathogens and the microbiological composition of the plaque taken from implant sites did not differ from that harvested from teeth. No significant differences were seen between implants and teeth for both pro- and anti-inflammatory cytokines. Even the histological examination showed no significant epithelial changes, although slight perivascular lymphocytic infiltration was seen in some biopsies.
Circadian Variations of Cortisol, Melatonin and Lymphocyte Subpopulations in Geriatric Age
Authors: Mazzoccoli G. , Vendemiale G. , La Viola M. , De Cata A. , Carughi S. , Greco A. , Balzanelli M. , Tarquini R.
Published: January 2017

A number of age-related changes in the 24-hour hormonal and non-hormonal rhythms have been found in older human beings. Lymphocyte subpopulations present circadian variation of some of their subsets and this variation may influence magnitude and expression of the immune responses. Numerous interactions exist among the nervous, endocrine and immune systems, mediated by neurotransmitters, hormones and cytokines. The aim of this study is to evaluate circadian variations of some endocrine and immune factors in older adults. Cortisol and melatonin serum levels were measured and lymphocyte subpopulation analyses were performed on blood samples collected every four hours for 24 hours from ten healthy young and middle-aged subjects and from ten healthy elderly subjects. There was a statistically significant difference between the groups in the observed values of CD20 (higher in young and middle-aged subjects) and CD25 and DR+ T cells (higher in elderly subjects). In the group of young and middle-aged subjects a clear circadian rhythm was validated for the time-qualified changes of all the factors studied. In the group of elderly subjects a number of rhythms were absent or altered. The results of the current study show that aging is associated with enhanced responsiveness of T cell compartment and alterations of circadian rhythmicity.
Thymoma-Associated Immunodeficiency: A Syndrome Characterized by Severe Alterations in NK, T and B-Cells and Progressive Increase in Naïve CD8+ T Cells
Authors: Vitiello L. , Masci A.M. , Montella L. , Perna F. , Angelini D.F. , Borsellino G. , Battistini L. , Merola G. , De Palma R. , Spadaro G. , Cosentini E. , Palmieri G. , Racioppi L.
Published: January 2017

Thymomas are rare tumours that sustain T-lymphopoiesis and trigger a variety of autoimmune diseases and immunodeficiencies, including a fatal hypogammaglobulinemia, namely Good's Syndrome (GS). Due to its rarity, GS has been poorly investigated and immunological features, as well as pathogenetic mechanisms underlying this syndrome, are unclear. We studied 30 thymoma patients by performing an immunological assessment, including immunophenotype and analysis of T cell repertoire (TCR). Development of GS was characterized by a progressive decrease in B, CD4 T and NK lymphocytes. These alterations paired with accumulation of CD8+CD45RA+ T cells that showed a polyclonal repertoire without expansions of specific clonotypes. GS is defined as hypogammaglobulinemia with thymoma. Here, we show for the first time that this syndrome is characterized by a severe loss of CD4+, NK and B cells. Furthermore, the accumulation of CD8+CD45RA+ T lymphocytes parallels these changes; this accumulation may have a role in determining the disease and can be used to monitor clinical stages of immunodeficiency in thymoma.
Giant Scrotal Elephantiasis: An Idiopathic Case
Authors: Dianzani C. , Gaspardini F. , Persichetti P. , Brunetti B. , Pizzuti A. , Margiotti K. , Degener A.M.
Published: January 2017

Scrotal elephantiasis is very rare disease in industrialized countries, where it is mainly due to surgery, irradiation or malignancies. It can be defined as idiopathic only when the possible congenital, infectious and compressive causes are excluded. We report a case of massive scrotal lymphoedema in an adult Caucasian patient, in Italy. He presented an extremely voluminous scrotal mass measuring 50 × 47 × 13 cm (weight 18 kg), which extended below his knees, invalidating all his daily activities. The patient was hospitalized in order to undergo to surgical treatment. Although genetic causes were searched and the possible role of infectious agents and compressive factors was evaluated, no etiology was ascertained. Histopathologic examination showed non-specific chronic inflammation, confirming the diagnosis of idiopathic elephantiasis. One year after surgical treatment, the patient is healthy without recurrence signs.
Role of Effector Cells (CCR7−CD27−) and Effector-Memory Cells (CCR7−CD27+) in Drug-Induced Maculopapular Exanthema
Authors: Fernandez T.D. , Torres M.J. , Lopez S. , Antunez C. , Gomez E. , Del Prado M.F. , Canto G. , Blanca M. , Mayorga C.
Published: January 2017

Maculopapular exanthema (MPE) induced by drugs is a T-cell mediated reaction and effector cells may play an important role in its development. We assessed the effector and cutaneous homing phenotype in peripheral blood cells from allergic patients after drug stimulation. This study included 10 patients and 10 controls. The effector phenotype (CCR7−CD27+/−), chemokine receptors (CCR4 and CCR10), and activation (CD25low) and regulatory markers (CD25high) were measured by flow cytometry in both peripheral blood mononuclear cells (PBMCs) and CD4-T-lymphocytes. Proliferation was determined by 5-(−6)-carboxyfluorescein diacetate succinimidyl ester (CFSE) assay and the migratory capacity by a chemotaxis assay using CCL17 and CCL27. Compared to controls, CCR7−CD27− cells were increased in patients without (p=0.003) and with drug stimulation (p<0.001) and had significantly higher proliferation (p=0.010). CCR10 expression was increased in patients after drug stimulation in total and memory CD27+ T-cells. Lymphocyte migration with CCL27 was higher in patients with drug stimulation (p=0.048), with a decrease in CCR7−CD27− (p<0.0001) and an increase in CCR7−CD27+ (p=0.017). In patients, CD4-T-lymphocytes were significantly activated after drug stimulation (p<0.001). In conclusion, we show that effector memory CD4+ T-cells (CCR7−CD27+) respond specifically to the drug responsible for MPE and confirm previous data about the involvement of CCR10 in cell trafficking to the skin.