Plaque-infiltrating T lymphocytes in patients with carotid atherosclerosis: an insight into the cellular mechanisms associated to plaque destabilization.

AIM: Inflammation plays a crucial role in the progression of atherosclerotic plaques. The aim of the present study was to investigate phenotypic and functional characteristics of plaque-infiltrating T lymphocytes associated with a complicated phenotype of carotid atherosclerotic lesions. METHODS: Atherosclerotic plaques were obtained from 17 patients undergoing carotid endarterectomy and cultured to isolate infiltrating T lymphocytes. Blood samples were obtained from patients and from 20 sex- and age-matched healthy subjects. The presence of lymphocytes (CD3+ cells) within atherosclerotic plaques was determined by immunohistochemistry. Phenotypic characteristics and intracellular cytokine expression of plaque-infiltrating and circulating T lymphocytes were determined by flow cytometry. Cytokine levels in supernatants from infiltrating T cell cultures were evaluated by enzyme-linked immunosorbent assay. RESULTS: A higher number of CD3+ cells was detected in complicated than in uncomplicated plaques. Complicated plaques had higher percentages of tumor necrosis factor (TNF)-α- and interferon (IFN)-γ- positive cells than uncomplicated ones, especially in CD4+ subpopulation. In patients the percentages of TNF-α-positive cells were higher in infiltrating than in circulating lymphocyte samples. Intracellular TNF-α, IFN-γ, interleukin (IL)-4 and IL-10 expression resulted higher in circulating lymphocyte samples from patients than in those from healthy subjects. Supernatants of infiltrating T cell cultures from complicated plaques showed higher levels of TNF-α and lower levels of IL-4 than those from uncomplicated plaques. CONCLUSION: Our data provide new information on the presence of increased percentages of pro-inflammatory T lymphocytes in complicated plaques with respect to uncomplicated ones and support the concept of the key role played by activated T cells in the progression of atherosclerotic lesions.

Identification of IP-10 and IL-5 as proteins differentially expressed in human complicated and uncomplicated carotid atherosclerotic plaques.

Inflammation plays a crucial role in the development and progression of atherosclerotic plaques. The aim of this study is to compare culture supernatants from uncomplicated and complicated carotid atherosclerotic plaques by a multiplex approach, to assess the molecular mediators associated with a plaque complicated phenotype. Atherosclerotic plaques were obtained from 17 patients undergoing carotid endarterectomy. Supernatants from plaque cultures were evaluated by Bio-Plex cytokine assay to determine 27 pro- and anti-inflammatory cytokines, chemokines and growth factors. Complicated plaques secreted higher levels of IP-10 (p = 0.027) and lower levels of IL-5 (p = 0.045) than did uncomplicated ones. Distinctive secretory patterns of cytokines, chemokines and growth factors were present in the two types of plaque. Our study identifies IP-10 and IL-5 as proteins differentiating complicated and uncomplicated plaques from human carotid arteries and provides new insights into the interplay of molecular mediators with atherosclerotic plaque progression.

Beta2-glycoprotein I is a target of T cell reactivity in patients with advanced carotid atherosclerotic plaques.

Evidence in animal models that beta2-glycoprotein I (beta2GPI), the principal target of autoimmune antiphospholipid antibodies, is involved in the initiation and progression of atherosclerosis, prompted us to investigate the possible role of this self protein as a target autoantigen of immune reactions in patients with carotid atherosclerosis. Plaque-infiltrating T lymphocytes from patients, and circulating T lymphocytes from patients and healthy subjects were tested by cell proliferation assay and by flow cytometry for intracellular cytokine expression in response to beta2GPI. ELISA was used to detect cytokine production in culture supernatants and anti-beta2GPI/anti-cardiolipin antibodies in serum samples. Eight of 35 PBMC samples and 1 of 5 plaque-infiltrating T lymphocyte samples from patients proliferated in response to beta2GPI, whereas PBMC from healthy subjects did not. Patients PBMC samples that proliferated in response to beta2GPI produced significantly higher IFN-gamma and TNF-alpha than non-proliferating PBMC. beta2GPI-specific plaque-derived T lymphocytes expressed IFN-gamma, TNF-alpha and IL-4, suggesting concomitant Th1 and Th2 activation. Only one patients serum was positive for anti-beta2GPI and anti-cardiolipin IgM antibodies. These new findings indicate that beta2GPI induces a cellular immune response in a subpopulation of patients with carotid atherosclerosis thus contributing to the inflammatory responses involved in carotid atherosclerotic disease.

Association of intracellular pro- and anti-inflammatory cytokines in peripheral blood with the clinical or ultrasound indications for carotid endarterectomy in patients with carotid atherosclerosis.

Early non-invasive diagnostic information would be useful in identifying patients at risk of progressive carotid atherosclerosis, despite an apparently harmless plaque on ultrasound imaging. In this study, we assessed the possible association of intracellular cytokines in peripheral blood with the ultrasound (stenosis > or = 70%) and clinical indications (transient ischaemic attack, amaurosis fugax or stroke) for carotid endarterectomy (CEA) in patients. Intracellular cytokine expression was determined in 106 patients (67 undergoing and 39 not undergoing CEA). Cells primed for the proinflammatory cytokines tumour necrosis factor (TNF)-alpha, interferon (IFN)-gamma, interleukin (IL)-1beta, IL-6, IL-8 and the anti-inflammatory cytokines IL-4 and IL-10 were found in significantly higher percentages in patients undergoing CEA than in patients who were not (P < 0.05). Intracellular cytokine expression was significantly higher in patients undergoing CEA who had stenosis > or = 70% (TNF-alpha, IFN-gamma, IL-1beta, IL-6, IL-4 and IL-10), with previous stroke (IFN-gamma, IL-1beta, IL-6, IL-8, IL-4 and IL-10) and with amaurosis fugax (IFN-gamma, IL-6, IL-4 and IL-10) than in patients not undergoing CEA. Increased intracellular cytokines in patients' peripheral blood might be a warning signal indicating progressive atherosclerosis. If so, intracellular cytokine monitoring could help in selecting patients at high risk of future clinical cardiovascular events and therefore most likely to benefit from CEA or adjustment of pharmacological therapy.

Morphological analysis of cell subpopulations within carotid atherosclerotic plaques.

Atherosclerosis is considered a chronic inflammatory process, prompted by lipid accumulation and propagated by cell-mediated mechanisms. The present work was undertaken to clarify this process by characterizing cellular components of inflammatory infiltrate localized within atheroma. Cryostat sections of atherosclerotic lesions obtained from human carotid endarterectomy were analysed immunohistochemically by using monoclonal and polyclonal antibody directed against T cell subpopulations (CD3, CD4, CD8), B cells (CD20), plasma cells (CD138), macrophages (CD14), mast cells (anti-tryptase). Our results assess that T cells are the predominant cell type among plaque infiltrating inflammatory cells. B cells were detected near the lipid core of atheroma and clusters of plasma cells were observed within cellular infiltrates in most plaques. Numerous tryptase positive mast cells were noticed in many areas of complicated lesions. Our results indicate the presence of many inflammatory cells within type V and VI atherosclerotic plaques, suggesting the involvement of those cells in plaque progression. In fact it was previously shown that stability of atherosclerotic lesions is influenced by mast cell-released matrix metalloproteinases which induce plaque rupture and by cytokines and chemokines which increase local inflammatory response and are produced by lymphocytes and macrophages.

Screening of a HUAEC cDNA library identifies actin as a candidate autoantigen associated with carotid atherosclerosis.

The humoral immune response to endothelium has a pivotal role in the development of atherosclerosis. Using a molecular method, we sought to identify endothelial autoantigens in carotid atherosclerosis. Immunoscreening of a HUAEC expression library with IgG from a pool of two sera from patients with carotid atherosclerosis identified a clone specific to actin. We evaluated actin-specific IgG reactivity in patients with carotid atherosclerosis and compared responses with those in patients with systemic lupus erythematosus and type 1 diabetes mellitus and in healthy subjects. Enzyme-linked immunoassay detected actin-specific IgG in a significantly higher percentage of sera from patients with atherosclerosis and systemic lupus erythematosus than from healthy subjects (16/61, 26% and 13/33, 39%versus 2/41, 5%, P = 0.012 and P < 10(-4), by chi2 test). Mean optical density values were significantly higher in patients than in healthy subjects (P < 10(-4) by Student's t-test). Patients with atherosclerosis and uncomplicated plaques had significantly higher serum anti-actin IgG reactivity than those with complicated plaques (P = 0.048 by Student's t-test). Our findings suggest that actin is an autoantigenic molecule of potential clinical interest in carotid atherosclerosis.

Cytokine expression in circulating T lymphocytes from patients undergoing carotid endarterectomy.

AIM: We investigated a possible relationship of cytokine expression and phenotype features of circulating T lymphocytes with the histological type of atherosclerotic plaque removed during carotid endarterectomy. METHODS: Peripheral blood samples were taken from 20 patients with carotid atherosclerosis and from 8 healthy blood donors. Patients were divided into 2 groups according to the histological type of their atherosclerotic plaques (types V and VI). Expression of intracellular tumor necrosis factor (TNF)-alpha, interferon (IFN)-gamma, interleukin-4 (IL-4), and of surface antigens (CD4, CD8, CD45RA, CD45RO, T cell receptor (TCR) alpha/beta, TCR gamma/delta) in circulating T lymphocytes was determined by 3-colour cytofluorimetric analysis. RESULTS: The percentage of T lymphocytes primed for TNF-alpha, IFN-gamma and IL-4 was higher in blood samples from patients than from healthy subjects; the difference was statistically significant for TNF-alpha-producing cells (p=0.01). In patients, the percentage of TNF-alpha-producing cells was significantly higher in the CD4+ subset than in the CD8+ subset (p=10(-4)). The percentage of TNF-alpha-, IFN-gamma- and IL-4-primed cells was higher in patients with type VI plaques (complicated lesions) than in patients with type V plaques (less complicated lesions). The difference was statistically significant for TNF-alpha-primed cells (p=0.05). No statistically significant differences were found in T cell phenotype features among patients or between patients and healthy subjects. CONCLUSION: Our results suggest a relationship between the percentage of circulating T lymphocytes expressing TNF-alpha and possibly IFN-gamma and IL-4 and the histological type of atherosclerotic plaque in patients with carotid artery disease.

[General anesthesia and cervical anesthesia. What changes in carotid surgery]. / Anestesia generale ed anestesia plessica. Cosa cambia in chirurgia carotidea.

Carotid surgery must be preventive; therefore cerebral protection procedures have been the centre of interest for a decade. Nowadays local cervical block anaesthesia seems to have changed the terms of the problem allowing achieve the aim of "no risk surgery". Therefore we considered our 16 year experience (352 carotids operated on 290 patients). Since 1990 we employed cervical block anaesthesia. In order to ratify as much as possible the two groups of comparison, considering that we adopted some exclusion criteria, we compared the results of the first hundred carotids that underwent surgery with general anaesthesia and the first hundred operated using cervical block anaesthesia. Since we adopted cervical block anaesthesia there was no need of intra-operative monitoring systems because we considered exclusively the patients' clinical answer to preclamping. The use of shunt decreased from 9% to 3%. Surgery performed in cervical block anaesthesia gives a positive impression. This is due to the fact that there is not only a significant reduction of the morbidity rate, especially from a neurologic point of view, but also a reduction of the post-operation hospital stay. Furthermore there is also a better organization of the surgical phases. Direct monitoring of the cerebral function allows a precise analysis of the peri-operatory neurological events. In conclusion our study suggests that cervical block anaesthesia allows clinical benefits for the patient as far as safety is concerned being also more convenient under the economic point of view.

[Intravascular surgery in the treatment of peripheral arteriopathies: our experience]. / La chirurgia endovascolare nel trattamento delle arteriopatie periferiche: la nostra esperienza.

The Authors, through a review of their vascular surgery experience in the treatment of PAOD at the III General Surgery Institute directed by Prof. G. Di Matteo (University, of Rome), focus their attention on endovascular surgery. Initially considered as an effective complement to "traditional surgery" rapidly its role has grown as an effective alternative for a number of vascular patients.

[Abdominal vascular and colonic pathology: the timing of combined treatment]. / Patologia vascolare addominale e colica: timing del trattamento combinato.

The authors report their experience with the combined treatment of concomitant vascular and colic pathology. After a wide review of the literature, the different surgical approaches to reduce the risks of mortality, morbidity and bacterial contamination of the vascular prosthesis are discussed.

[Abdominal vascular and biliary pathology: timing of the combined treatment]. / Patologia vascolare addominale e biliare: timing del trattamento combinato.

The Authors, after an accurate review of the Literature, report their experience in the combined management of aorto-iliac and biliary lesions. Surgical priorities are well specified in order to reduce the possibility of bacterial contamination of the vascular prosthesis.

[The changes in the sexual behavior of patients undergoing aorto-iliac revascularization]. / Studio delle modificazioni dell'attività sessuale nei pazienti sottoposti a rivascolarizzazione aorto-iliaca.

A correct approach to sexual disorders in vascular patients presupposes an accurate investigation before and after operation. The authors evaluate the reliability of the diagnostic methods used, pre- and postoperatively, to ascertain sexual disorders, and analyse the influence of surgery on sexual function.

[Primary "small-cell" lymphoma of the lung. A clinical case report]. / Linfoma primitivo del polmone "a piccole cellule". Osservazioni su un caso clinico.

Primary lymphoma of the lung is a rare pathological condition arising from mucosa-associated lymphoid tissue (MALT). The lack of specific symptoms and the related diagnostic problems induced the authors to report a clinical case recently observed. Therefore, some histopathologic characteristics useful for a correct differential diagnosis with the pseudolymphoma of the lung and the interstitial lymphocyte pneumonia are analysed.