Tryptophan-derived Catabolites Are Responsible for Inhibition of T and Natural Killer Cell Proliferation Induced by Indoleamine 2,3-Dioxygenase

Macrophages exposed to macrophage colony-stimulating factor acquire the capacity to suppress T cell proliferation; this effect is associated with de novo expression of the tryptophan-catabolizing enzyme indoleamine 2,3-dioxygenase (IDO). We have purified IDO and tested its activity in in vitro models of T cell activation. IDO was able to inhibit proliferation of CD4+ T lymphocytes, CD8+ T lymphocytes, and natural killer (NK) cells; proliferation of B lymphocytes was not affected. The inhibitory role of tryptophan and of its catabolites was then tested. In the presence of tryptophan, only l-kynurenine and picolinic acid inhibit cell proliferation. In a tryptophan-free medium cell proliferation was not affected. In the absence of tryptophan inhibition induced by l-kynurenine and picolinic acid was observed at concentrations below the lowest concentration that was effective in the presence of tryptophan, and quinolinic acid acquired some inhibitory capacity. Inhibition of cell proliferation induced by the tryptophan catabolites resulting from IDO activity was selective, applying only to cells undergoing activation. Resting cells were not affected and could subsequently activate normally. We suggest that IDO exerts its effect on cell proliferation by (i) starting the cascade of biochemical reactions that produce the three catabolites and by (ii) enhancing their inhibitory potential by depriving the extracellular microenvironment of tryptophan

High Human Papillomavirus DNA loads in Inflammatory Middle Ear Diseases

Background. Previous studies reported human papillomaviruses (HPVs) in middle ear tumors, whereas these viruses have been poorly investigated in chronic inflammatory middle ear diseases. The purpose of this study was to investigate HPVs in non-tumor middle ear diseases, including chronic otitis media (COM). Methods. COM specimens (n=52), including chronic suppurative otitis media (CSOM) (n=38) and cholesteatoma (COMC) (n=14), as well as normal middle ear specimens (NME) (n=56) were analyzed. HPV DNA sequences and DNA loads were analyzed by quantitative PCR. HPV genotyping was performed by direct sequencing of the amplimers. Results. HPV DNA was detected in 23% (12/52) of COM and in 30.4% (17/56) NME (p>0.05). Specifically, HPV DNA sequences were revealed in 26.3% (10/38) of CSOM and in 14.3% (2/14) COMC (p>.05). Interestingly, the HPV DNA load was higher in COMC (mean 7.47 copy/cell) than in CSOM (mean 1.02 copy/cell), and NME (mean 1.18 copy/cell) (P=.03 and P=.017 versus CSOM and NME, respectively). HPV16 and HPV18 were the main genotypes detected in COMC, CSOM and NME. Conclusions. This data indicates that HPV-positive CSOM and COMC are generally associated with higher viral DNA loads as compared to NME. In addition, for the first time, HPVs were detected in normal middle ear mucosa specimens. This result suggests that NME is an additional epithelial tissue that can be HPV infected

Social cognition impairment in genetic frontotemporal dementia within the GENFI cohort

A key symptom of frontotemporal dementia (FTD) is difficulty interacting socially with others. Social cognition problems in FTD include impaired emotion processing and theory of mind difficulties, and whilst these have been studied extensively in sporadic FTD, few studies have investigated them in familial FTD. Facial Emotion Recognition (FER) and Faux Pas (FP) recognition tests were used to study social cognition within the Genetic Frontotemporal Dementia Initiative (GENFI), a large familial FTD cohort of C9orf72, GRN, and MAPT mutation carriers. 627 participants undertook at least one of the tasks, and were separated into mutation-negative healthy controls, presymptomatic mutation carriers (split into early and late groups) and symptomatic mutation carriers. Groups were compared using a linear regression model with bootstrapping, adjusting for age, sex, education, and for the FP recognition test, language. Neural correlates of social cognition deficits were explored using a voxel-based morphometry (VBM) study. All three of the symptomatic genetic groups were impaired on both tasks with no significant difference between them. However, prior to onset, only the late presymptomatic C9orf72 mutation carriers on the FER test were impaired compared to the control group, with a subanalysis showing differences particularly in fear and sadness. The VBM analysis revealed that impaired social cognition was mainly associated with a left hemisphere predominant network of regions involving particularly the striatum, orbitofrontal cortex and insula, and to a lesser extent the inferomedial temporal lobe and other areas of the frontal lobe. In conclusion, theory of mind and emotion processing abilities are impaired in familial FTD, with early changes occurring prior to symptom onset in C9orf72 presymptomatic mutation carriers. Future work should investigate how performance changes over time, in order to gain a clearer insight into social cognitive impairment over the course of the disease

Detection of Merkel Cell Polyomavirus DNA in Serum Samples of Healthy Blood Donors

Merkel cell polyomavirus (MCPyV) has been detected in 80% of Merkel cell carcinomas (MCC). In the host, the MCPyV reservoir remains elusive. MCPyV DNA sequences were revealed in blood donor buffy coats. In this study, MCPyV DNA sequences were investigated in the sera (n = 190) of healthy blood donors. Two MCPyV DNA sequences, coding for the viral oncoprotein large T antigen (LT), were investigated using polymerase chain reaction (PCR) methods and DNA sequencing. Circulating MCPyV sequences were detected in sera with a prevalence of 2.6% (5/190), at low-DNA viral load, which is in the range of 1–4 and 1–5 copies/μl by real-time PCR and droplet digital PCR, respectively. DNA sequencing carried out in the five MCPyV-positive samples indicated that the two MCPyV LT sequences which were analyzed belong to the MKL-1 strain. Circulating MCPyV LT sequences are present in blood donor sera. MCPyV-positive samples from blood donors could represent a potential vehicle for MCPyV infection in receivers, whereas an increase in viral load may occur with multiple blood transfusions. In certain patient conditions, such as immune-depression/suppression, additional disease or old age, transfusion of MCPyV-positive samples could be an additional risk factor for MCC onset

Merkel cell carcinomas arising in autoimmune disease affected patients treated with biologic drugs including anti-TNF

open12siThis study has been supported in parts by Associazione Italiana per la Ricerca sul Cancro (AIRC), grant IG16046; Associazione Sammarinese per la Lotta contro le Leucemie e le Emopatie Maligne (ASLEM), grant CFR/2015; LIONS Club International, District 108 TB, Italy, grant UNIFE 2015; and Fondazione Cassa di Risparmio di Cento, grant 2014 (all to M. Tognon).Purpose: The purpose of this investigation was to characterize Merkel cell carcinomas (MCC) arisen in patients affected by autoimmune diseases and treated with biologic drugs. Experimental Design: Serum samples from patients with MCC were analyzed for the presence and titer of antibodies against antigens of the oncogenic Merkel cell polyomavirus (MCPyV). IgG antibodies against the viral oncoproteins large T (LT) and small T (ST) antigens and the viral capsid protein-1 were analyzed by indirect ELISA. Viral antigens were recombinant LT/ST and virus-like particles (VLP), respectively. MCPyV DNA sequences were studied using PCR methods in MCC tissues and in peripheral blood mononuclear cells (PBMC). Immunohistochemical (IHC) analyses were carried out in MCC tissues to reveal MCPyV LT oncoprotein. Results: MCPyV DNA sequences identified in MCC tissues showed 100% homology with the European MKL-1 strain. PBMCs from patients tested MCPyV-negative. Viral DNA loads in the three MCC tissues were in the 0.1 to 30 copy/cell range. IgG antibodies against LT/ST were detected in patients 1 and 3, whereas patient 2 did not react to the MCPyV LT/ST antigen. Sera from the three patients with MCC contained IgG antibodies against MCPyV VP1. MCC tissues tested MCPyV LT-antigen–positive in IHC assays, with strong LT expression with diffuse nuclear localization. Normal tissues tested MCPyV LT–negative when employed as control. Conclusions: We investigated three new MCCs in patients affected by rheumatologic diseases treated with biologic drugs, including TNF. A possible cause–effect relationship between pharmacologic immunosuppressive treatment and MCC onset is suggested. Indeed, MCC is associated with MCPyV LT oncoprotein activity.openRotondo, John Charles; Bononi, Ilaria; Puozzo, Andrea; Govoni, Marcello; Foschi, Valentina; Lanza, Giovanni; Gafa, Roberta; Gaboriaud, Pauline; Touzã©, Françoise Antoine; Selvatici, Rita; Martini, Fernanda; Tognon, MauroRotondo, John Charles; Bononi, Ilaria; Puozzo, Andrea; Govoni, Marcello; Foschi, Valentina; Lanza, Giovanni; Gafa, Roberta; Gaboriaud, Pauline; Touzã©, Françoise Antoine; Selvatici, Rita; Martini, Fernanda; Tognon, Maur

Extra-pleuric coaxial system for CT-guided percutaneous fine-needle aspiration biopsy (FNAB) of small (≤20 mm) lung nodules: a novel technique using multiplanar reconstruction (MPR) images

The aim of the study is to present the diagnostic feasibility, usefulness, and safety of a novel technique for coaxial CT-guided fine-needle aspiration biopsy of small (≤20 mm in diameter) lung nodules. A 18-gauge (G) (1.2 × 40 mm) needle is inserted through the skin in the depth of the thoracic wall tissues remaining outside the pleura. Its positioning is planned and adjusted using multiplanar reconstruction (MPR) images along the 18-G guide needle axis tracing a reference outline extended from the needle tip to the target nodule. When the insertion of the 18-G extra-pleuric needle (EPN) proves to be precise, a 22-G Chiba needle is then passed through the outer 18-G EPN until it reaches the thoracic lesion for the sampling procedure. Patient population included 153 males and 94 females, with a mean age of 61.3 ± 21.6 years. Mean nodule diameter was 14.1 ± 2.2 mm. The lesion depth from pleural plane ranged from 0 mm to 127 mm. An average of 1.29 aspirates were performed per lesion. The most common complication was pneumothorax in 27 cases; there were no cases of PNX requiring chest tube insertion. Intrapulmonary bleeding along the needle track was observed in 32 patients. Exploiting the advantage of MPR images, our novel technique of extra-pleuric coaxial system with a 18-G EPN allows the operator to multiple samplings of small (≤20 mm) target lesions in various locations with a thinner (22-G Chiba) needle, thus reducing the degree of pleural, parenchymal, or adjacent organs damage