Giancarlo Torre

CD8+CD28− T Regulatory Lymphocytes Inhibiting T Cell Proliferative and Cytotoxic Functions Infiltrate Human Cancers

Tumor growth is allowed by its ability to escape immune system surveillance. An important role in determining tumor evasion from immune control might be played by tumor-infiltrating regulatory lymphocytes. This study was aimed at characterizing phenotype and function of CD8+CD28− T regulatory cells infiltrating human cancer. Lymphocytes infiltrating primitive tumor lesion and/or satellite lymph node from a series of 42 human cancers were phenotypically studied and functionally analyzed by suppressor assays. The unprecedented observation was made that CD8+CD28− T regulatory lymphocytes are almost constantly present and functional in human tumors, being able to inhibit both T cell proliferation and cytotoxicity. CD4+CD25+ T regulatory lymphocytes associate with CD8+CD28− T regulatory cells so that the immunosuppressive activity of tumor-infiltrating regulatory T cell subsets, altogether considered, may become predominant. The infiltration of regulatory T cells seems tumor related, being present in metastatic but not in metastasis-free satellite lymph nodes; it likely depends on both in situ generation (via cytokine production) and recruitment from the periphery (via chemokine secretion). Collectively, these results have pathogenic relevance and implication for immunotherapy of cancer.

A model to prioritize access to elective surgery on the basis of clinical urgency and waiting time

BackgroundPrioritization of waiting lists for elective surgery represents a major issue in public systems in view of the fact that patients often suffer from consequences of long waiting times. In addition, administrative and standardized data on waiting lists are generally lacking in Italy, where no detailed national reports are available. This is true although since 2002 the National Government has defined implicit Urgency-Related Groups (URGs) associated with Maximum Time Before Treatment (MTBT), similar to the Australian classification. The aim of this paper is to propose a model to manage waiting lists and prioritize admissions to elective surgery.MethodsIn 2001, the Italian Ministry of Health funded the Surgical Waiting List Info System (SWALIS) project, with the aim of experimenting solutions for managing elective surgery waiting lists. The project was split into two phases. In the first project phase, ten surgical units in the largest hospital of the Liguria Region were involved in the design of a pre-admission process model. The model was embedded in a Web based software, adopting Italian URGs with minor modifications. The SWALIS pre-admission process was based on the following steps: 1) urgency assessment into URGs; 2) correspondent assignment of a pre-set MTBT; 3) real time prioritization of every referral on the list, according to urgency and waiting time. In the second project phase a prospective descriptive study was performed, when a single general surgery unit was selected as the deployment and test bed, managing all registrations from March 2004 to March 2007 (1809 ordinary and 597 day cases). From August 2005, once the SWALIS model had been modified, waiting lists were monitored and analyzed, measuring the impact of the model by a set of performance indexes (average waiting time, length of the waiting list) and Appropriate Performance Index (API).ResultsThe SWALIS pre-admission model was used for all registrations in the test period, fully covering the case mix of the patients referred to surgery. The software produced real time data and advanced parameters, providing patients and users useful tools to manage waiting lists and to schedule hospital admissions with ease and efficiency. The model protected patients from horizontal and vertical inequities, while positive changes in API were observed in the latest period, meaning that more patients were treated within their MTBT.ConclusionThe SWALIS model achieves the purpose of providing useful data to monitor waiting lists appropriately. It allows homogeneous and standardized prioritization, enhancing transparency, efficiency and equity. Due to its applicability, it might represent a pragmatic approach towards surgical waiting lists, useful in both clinical practice and strategic resource management.

Clonal Analysis of T Lymphocytes Infiltrating the Thyroid Gland in Hashimoto’s Thyroiditis

T cells isolated from thyroid tissue and peripheral blood of 2 patients with Hashimotos thyroiditis were studied by a high cloning efficiency microculture technique. Clonal efficiencies of 37 and 24% were obtained from thyroid-derived T cell cultures, while 40 and 90% efficiencies resulted from peripheral-blood-derived cultures. A prevalence of T4-/T8+ T cell clones were found in thyroid infiltrates. The functional analysis of the clones demonstrated significantly higher proportions of clones with cytolytic activity in a lectin-dependent assay in thyroid-derived microcultures, as compared to peripheral blood-derived ones. The proportion of clones displaying natural-killer-like activity was increased in 1 patient only. Cytolytic activity was displayed not only by all T4-/T8+, but also by several T4+/T8- intrathyroid clones. Remarkable proportions of cytolytic clones were also able to release interleukin-2 upon phytohemagglutinin stimulation. Finally, the proportion of T cell clones able to release gamma-interferon following mitogen stimulation was significantly higher in thyroid- vs. peripheral-blood-derived microcultures. These results provide further data about the possible pathogenetical role of both regulatory and effector T lymphocytes in human autoimmune thyroiditis.

Non‐Antigen‐Specific CD8+ T Suppressor Lymphocytes in Diseases Characterized by Chronic Immune Responses and Inflammation

Abstract: Recent studies on regulatory lymphocytes demonstrate that CD8+ T suppressor (Ts) cells may have great relevance in controlling immune system homeostasis and avoiding development of chronic inflammatory diseases. Among the three subpopulations of CD8+ Ts cells so far recognized in humans, the type 2 (non‐antigen‐specific) cell is characterized by the capacity to inhibit both T cell proliferation and cytotoxic T lymphocyte activity through secretion of soluble factors. Previous work has shown the impairment of in vitro generation of type 2 CD8+ Ts cells from the peripheral blood of relapsed patients with multiple sclerosis, systemic lupus erythematosus, or systemic sclerosis. Here, similar findings are demonstrated for patients with human immunodeficiency virus or chronic hepatitis C virus infection. Furthermore, the presence of type 2 CD8+ Ts cells infiltrating diseased tissues in patients with autoimmune thyroiditis or cancer is shown. Collectively, these findings suggest that type 2 CD8+ Ts cells may be involved in the control of pathologic chronic immune responses, contributing in some cases to the pathogenesis of the disease.

Different intrathyroid expression of intercellular adhesion molecule-1 (ICAM-1) in Hashimoto's thyroiditis and Graves' disease: Analysis at mRNA level and association with B7.1 costimulatory molecule

Cultured thyroid epithelial cells can be induced to express intercellular adhesion molecule-1 (ICAM-1, or CD54). However, constitutive follicular expression of ICAM-1 has been reported only in thyroid autoimmunity. We evaluated the expression of ICAM-1 mRNA and protein on thyroid tissue from different autoimmune thyroid diseases, and its relationship with other immunologically relevant surface markers, namely costimulatory molecules of B7 family. Thyroid tissue sections were obtained by surgically removed thyroid glands from 6 patients with Hashimoto’s thyroiditis (HT), 6 with Graves’ disease (GD) and 3 with multinodular nontoxic goiter. We used in situ hybridization to localize ICAM-1 mRNA, and immunohistochemical analysis by alkaline phosphatase anti-alkaline phosphatase (APAAP) method. We showed a clear hybridization pattern, localized in follicular cells, in sections of glands with HT. The hybridization pattern was far less pronounced in GD: no staining was apparent on follicular cells. These results were strictly consistent with those obtained by means of immunohistochemistry. Moreover, double-staining experiments demonstrated colocalization of ICAM-1 and B7.1 molecules in HT, whereas no B7.1 expression was observed in Graves’ or in non-autoimmune thyroid diseases. These data agree with the hypothesis of distinct immunoregulatory phenomena and effector mechanisms in the 2 main autoimmune thyroid diseases.

Fibrous invasive (Riedel's) thyroiditis with critical response to steroid treatment.

The Riedel’s thyroiditis is an uncommon form of chronic thyroiditis characterized by an invasive fibrosclerosis of the gland, often involving the surrounding tissues. Usually, the only possible treatment is the surgical decompression of the tissues. We describe a case of aggressive Riedel’s thyroiditis with severe compression and dislocation of thrachea and esophagus. The surgical approach was uneffective, while an “ex juvantibus” steroid treatment, resulted in a dramatic regression of fibrosclerosis and a complete clinical remission. This report points out the possible effectiveness of corticosteroids in the treatment of selective disorders involving increased fibrogenesis.

The role of simulation in developing communication and gestural skills in medical students

BackgroundInternational studies have shown that laboratory training, particularly through the application of the principles of simulation learning, is an effective means of developing the communication and gestural skills of healthcare professionals. At the Advanced Simulation Center of the University of Genoa we have therefore established the first clinical skill laboratory with medical school students and an interprofessional team of trainers, as the first step towards developing simulation training of both medical and nursing students at our University.The aim of this study was to assess student satisfaction with laboratory training in an Advanced Simulation Center.MethodsAll of the third-year students of the Medical School (n = 261) were invited to participate in the laboratory sessions at the Advanced Simulation Center. They were divided into groups and attended the Center for one week. The team of trainers included medical doctors and nurses involved in teaching at the University Medicine and Nursing programs. At the end of the week, the students were administered an anonymous questionnaire made up of two sections: the first one was on the content of individual laboratory sessions; the second on the training methods, materials used and the trainers. A five-point Likert scale was used to measure satisfaction.ResultsAccording to the students all of the topics covered by the laboratory sessions were irreplaceable. Questionnaire results showed a high level of satisfaction with the methods used, the instruments developed, and with the expertise and approachability of the educators. Almost all of the students wanted to participate in similar laboratory activities in the future.ConclusionsThe study highlighted the need to permanently integrate laboratory training sessions into the curriculum of medical students, who found them very useful and stimulating. The limit of this study was that only the teaching staff was interprofessional, and the students were only 3rd Year students of medicine.In the future, we hope to include also nursing students because they will need to learn how to deal with aspects of their clinical practice that require an interprofessional approach.

Expression of very late activation antigen-1 on intrathyroid lymphocytes in autoimmune thyroid disease

Very late activation antigen-1 (VLA-1) is a ß1-integrin implied in interaction with extracellular matrix components. It is expressed by T lymphocytes upon prolonged activation in vitro. In this work we have evaluated VLA-1 expression in thyroid infiltrates and peripheral blood of patients with autoimmune thyroid disease (AITD), by immunofluorescence on dispersed cells and by in situ analysis on frozen tissue sections. The results obtained showed increased proportions of VLA-1 positive lymphocytes in thyroid infiltrates, similarly to that observed with “early” activation antigens. No positivity of thyroid follicular cells was observed. Given the role of the VLA-1 molecule in lymphocyte adhesion, the increased VLA-1 positivity of infiltrating lymphocytes is consistent with intrathyroidal homing of long-term activated cells, possibly relevant to AITD pathogenesis.

Is euthyroidism the goal of surgical treatment of diffuse toxic goitre

OBJECTIVE To find out by studying a homogeneous group of patients whether euthyroidism is achievable by surgical treatment of diffuse toxic goitre. DESIGN Retrospective study. SETTING Teaching hospital, Italy. SUBJECTS 128 of the 152 patients operated on for diffuse toxic goitre during the period January 1971-December 1994 and followed up for a median of 83 months (range 6-289). INTERVENTION. Standard subtotal thyroidectomy. MAIN OUTCOME MEASURES Operative mortality, recurrence, hypothyroidism and late complications. RESULTS There were no operative deaths. After 10 years follow up, 11 patients (9%) had developed recurrences and 61 (48%) were euthyroid. In the univariate analysis the risk of hypothyroidism was significantly associated with the year of operation (p = 0.04), the duration of symptoms (p < 0.01), and the degree of lymphocytic infiltration (p < 0.01). The last two were confirmed by multivariate analysis. CONCLUSION Subtotal thyroidectomy seems to be an effective treatment of diffuse toxic goitre as a stable euthyroid state can be achieved in nearly half the patients after a prolonged follow up.

Differentiated thyroid cancer: surgical treatment of 190 patients

Abstract Between 1968 and 1991, 190 patients (51 men, 139 women) with a mean age of 46.3 years underwent surgery for differentiated thyroid cancer (148 papillary and 42 follicular carcinomas). In 29.5% of the cases a concomitant goitre was histologically demonstrated. These patients were significantly older (mean: 54.7 years) (P 40 years), pT, stage, pM and symptomatic metastases.