Gianluca Taccagni

Human Papillomavirus Infection and Associated Cervical Disease in Human Immunodeficiency Virus-Infected Women: Effect of Highly Active Antiretroviral Therapy

To determine the effect of highly active antiretroviral therapy (HAART) on high-risk human papillomavirus (HR-HPV) infections and related cervical lesions, the virologic and cytologic markers of HPV infection were prospectively studied in 163 human immunodeficiency virus (HIV)-infected women, including 27 untreated, 62 treated with reverse transcriptase inhibitors, and 74 treated with HAART. A high prevalence of both infections with HR-HPV types (68%) and squamous intraepithelial lesions (SILs; low grade, 20.2%; high grade, 6.2%) was observed. The risks of infection and disease were inversely correlated with CD4 cell counts (P=.015 and P=.022, respectively). During the observation period (mean, 15.4 months; range, 6-24 months), CD4 cell counts increased significantly only in subjects receiving HAART (P<.001). Persistence of HR-HPV infection and progression of SILs were comparable in the 3 groups. These results indicate that, even in the era of HAART, HIV-infected women should be monitored carefully for the emergence of high-grade SILs and cervical cancer.

Solitary fibrous tumour of the thyroid: clinicopathological, immunohistochemical and ultrastructural study of three cases.

We describe three cases of solitary fibrous tumour (SFT) arising from thyroid stroma. Grossly, the tumours were clearly delimited but only partly encapsulated. The following histomorphological growth patterns were observed: bundles of cells in storiform configuration; non-structured bundles; prevalence of fibrous matrix; highly cellular, non-structured; prevalence of loose, non-structured extracellular substance; cellular proliferation and vascular spaces in a haemangiopericytic configuration and a lipomatous component. Immunohistochemical investigation demonstrated intense, diffuse vimentin positivity and focal, less intense actin positivity in all three cases. At electron microscopy we observed a primitive cell of mesenchymal type, with cytoplasm poor in organelles and rich in filaments; this cell sometimes presented differentiation characteristics. SFT is at present the most correct term for the lesions presented here despite some morphological characteristics which differ from cases reported in the literature.

Human vasculogenesis ex vivo: embryonal aorta as a tool for isolation of endothelial cell progenitors.

Vasculogenesis, the de novo formation of new blood vessels from undifferentiated precursor cells or angioblasts, has been studied with experimental in vivo and ex vivo animal models, but its mechanism is poorly understood, particularly in humans. We used the aortic ring assay to investigate the angioforming capacity of aortic explants from 11- to 12-week-old human embryos. After being embedded in collagen gels, the aorta rings produced branching capillary-like structures formed by mesenchymal spindle cells that lined a capillary-like lumen and expressed markers of endothelial differentiation (CD31, CD34, von Willebrand factor [vWF], and fms-like tyrosine kinase-1 [Flk-1;[sol;vascular endothelial growth factor receptor 2 [VEGFR2]). The cell linings of these structures showed ultrastructural evidence of endothelial differentiation. The neovascular proliferation occurred primarily in the outer aspects of aortic rings, thus suggesting that the new vessels mainly arose from immature endothelial precursor cells localized in the outer layer of the aortic stroma, ie, a process of vasculogenesis rather than angiogenesis. The undifferentiated mesenchymal cells (CD34+/CD31−), isolated and cultured on collagen-fibronectin, differentiated into endothelial cells expressing CD31 and vWF. Furthermore, the CD34+/CD31+ cells were capable of forming a network of capillary-like structures when cultured on Matrigel. This is the first reported study showing the ex vivo formation of human microvessels by vasculogenesis. Our findings indicate that the human embryonic aorta is a rich source of CD34+/CD31− endothelial progenitor cells (angioblasts), and this information may prove valuable in studies of vascular regeneration and tissue bioengineering.

Myofibrosarcoma of the breast: review of the literature on myofibroblastic tumors and criteria for defining myofibroblastic differentiation.

A case of myofibrosarcoma of the breast is described. A 55-year-old woman presented with a small mammary nodule. A large recurrent lesions appeared a month later, and the patient died 11 months following initial presentation from diffuse pleuropulmonary metastases. Histologically, the primary tumor consisted mainly of spindled cells, arranged in fascicles and surrounded by varying quantities of dense hyaline collagen. The recurrent lesion had a more pleomorphic organization. In both lesions, there was positive immunostaining for vimentin, smooth-muscle actin, and fibronectin, and negative results for desmin, laminin, and type IV collagen. Electron microscopy revealed abundant rough endoplasmic reticulum, myofilaments with focal densities, and the fibronexus junctions and fibronectin fibrils characteristic of myofibroblasts. Given these cellular features and behavior, the tumor was interpreted as a malignant neoplasm showing myofibroblastic differentiation, i.e., a myofibrosarcoma. This case enlarges the group of myofibrosarcoma of breast, also with the demonstration of fibronexus and fibronectin fibrils. The paper emphasizes the criteria required for myofibroblastic differentiation and reviews lesions of the breast reported in the literature as myofibroblastic.

Productive HIV-1 infection of human cervical tissue ex vivo is associated with the secretory phase of the menstrual cycle.

Cervical tissue explants (CTEs) from 22 HIV-1 seronegative women were exposed to R5 HIV-1 ex vivo. Eight CTEs were productively infected in terms of HIV-1 p24Gag release in culture supernatants, whereas 14 were not. Nonetheless, both accumulation of HIV-1gag DNA and of p24Gag+ CD4+ T cells and macrophages occurred in both productive and, at lower levels, in nonproductive CTEs. Nonproductive CTEs differed from productive CTEs for higher secretion of C-C motif chemokine ligand 3 (CCL3) and CCL5. A post-hoc analysis revealed that all productive CTEs were established from women in their secretory phase of the menstrual cycle, whereas nonproductive CTEs were derived from women either in their secretory (28%) or proliferative (36%) menstrual cycle phases or with an atrophic endometrium (36%). Thus, our results support the epidemiological observation that sexual HIV-1 transmission from males to women as well as from women to men is more efficient during their secretory phase of the menstrual cycle.

Determination of Human Papillomavirus (HPV) Load and Type in High-Grade Cervical Lesions Surgically Resected from HIV-Infected Women during Follow-up of HPV Infection

BACKGROUND The role of human papillomavirus (HPV) load and the importance of multiple-strain HPV infections as biomarkers for the development of cervical disease were evaluated in human immunodeficiency virus (HIV)-positive women. METHODS A total of 108 samples were analyzed, 64 of which were obtained from 16 HIV-positive women who underwent surgical resection of the cervical cone for treatment of a histologically confirmed high-grade cervical intraepithelial neoplasm (cases) and 44 of which were obtained from 22 HIV-positive women who had high-risk HPV but a negative colposcopy result (controls). Each patient underwent periodic examinations at 6-12-month intervals that included colposcopy, Papanicolaou testing, biopsy (if indicated), and cervical brushing for HPV testing. Viral typing was performed by reverse dot-blot hybridization and quantification of viral load by in-house real-time PCR and commercial assays. RESULTS Analysis of the cervical-brush samples collected when high-grade squamous intraepithelial lesions were diagnosed revealed that all cases had HPV loads that were significantly higher than those of controls (P=.0004 and P=.0003, by PCR and the Hybrid Capture 2 index [Digene], respectively). Decreasing concentrations of HPV load were observed when comparing samples obtained before and after treatment (P<.0001). The number and type of HPV strains that were detected were not statistically different between cases and controls. CONCLUSIONS The significantly higher HPV load detected in women with high-grade cervical dysplasia, as well as the dramatic decrease in the load after surgical removal of the lesion, suggest that HPV load is a possible prognostic marker of high-grade SIL.

IFN-γ Produced by Human Papilloma Virus-18 E6-Specific CD4+ T Cells Predicts the Clinical Outcome after Surgery in Patients with High-Grade Cervical Lesions

Cervical neoplastic lesions are associated with infection by high-risk human papilloma viruses (HPVs). HPV-16 and HPV-18 are the most common genotypes. It has been proposed that development of HPV-16-positive cervical lesions is associated with impaired CD4+ T cell immunity against early Ags. The aim of the study was to evaluate whether this impairment also applies to HPV-18. We investigated the presence and the quality of anti-HPV-18 E6 CD4+ T cell responses in the blood of 37 consecutive patients with high-grade cervical lesions, 25 normal donors, and 20 cord bloods. The immune infiltrate in the cervical lesions was also evaluated. The characteristics of the responses were correlated to the clinical outcome. We found that one or more HPV-18 E6 peptides, containing naturally processed epitopes, were able to induce a response in 40–50% of the patients, depending on the effector function tested. Importantly, these percentages rose to 80–100% when HPV-18-positive patients were considered. HPV-18 E6-specific CD4+ T cells produced mixed Th1/Th2 responses and statistical analysis of the cytokines produced revealed that the amount of IFN-γ released could predict infection persistence and/or disease relapse after surgery. Finally, we found that a higher number of infiltrating CD4+ and T-bet+ T cells in the lesions correlated with a favorable clinical outcome. Our results strongly suggest a relevant role for CD4+ T cells in the control of the HPV-18 compared with HPV-16 infections in patients with high-grade cervical lesions and identify an immunologic parameter potentially useful for patients’ stratification.

Unraveling the two entities of endometrioid ovarian cancer: A single center clinical experience

OBJECTIVE Due to the increasing prevalence of the benign condition, ovarian carcinoma arising from endometriosis is emerging as a relevant clinical entity with an unclear biological signature. We have investigated clinical and histologic features of endometriosis-associated endometrioid ovarian cancer using an institutional retrospective database. METHODS Patients diagnosed with endometrioid ovarian cancer at our institution were divided into two groups according to the fulfillment or not of Sampsons and Scotts criteria for the detection of endometriosis-associated ovarian cancer. Clinical and histological data were reported and compared. Survival analysis was obtained using the log-rank test in an unadjusted Kaplan-Meier method. Multivariate analysis was performed using the Cox proportional hazards regression model to establish independent factors associated with endometriosis-associated endometrioid ovarian cancer and to identify predictors of survival. The degree of concordance was evaluated by Cohens Kappa measures. RESULTS Patients with endometriosis-associated endometrioid ovarian cancer were significantly younger, had a lower disease stage (62% vs 23%; p=0.003), a less prevalent high grade tumor (38% vs 82%; p=0.002) and a higher prevalence of squamous and mucinous metaplasia. The rate of endometrial cancer diagnosis was significantly higher in women with endometriosis-associated endometrioid ovarian cancer (33%) than in other patients (11%) (p=0.04) with a 92% concordance between ovarian and endometrial histologic tumor grade. A significant difference in survival rate could not be demonstrated between patients with or without endometriosis. CONCLUSIONS The analysis of a retrospective endometrioid ovarian cancer database may allow to suggest a 40 molecular, morphological and clinical parallelism between endometrial and endometrioid ovarian cancers.

[18f]fluorodeoxyglucose positron emission tomography as a useful indicator of metastatic gestational trophoblastic tumor: preliminary results in three patients

OBJECTIVE The goal of this study was to evaluate the usefulness of positron emission tomography with [(18)F]fluorodeoxyglucose ([(18)F]FDG-PET) in detecting metastases in patients with gestational trophoblastic tumor (GTTs). METHODS A retrospective study was conducted on three patients with GTTs who had been studied with [(18)F]FDG-PET and computed tomography (CT) after an increase in human chorionic beta-gonadotropin (betahCG) serum levels. PET scans were performed with a multiring whole-body positron emission tomograph 45 min after an intravenous bolus injection of [(18)F]FDG ( approximately 5.2 MBq/kg). CT studies were obtained on a spiral scanner prior and after administration of intravenous iodinated contrast material. Within a week of CT and [(18)F]FDG-PET studies, the patients underwent surgical procedures for histological diagnosis. RESULTS In one patient, a lung lesion positive for neoplastic tissue with [(18)F]FDG-PET and negative with CT was confirmed to be a GTT metastasis at histology. In another patient, [(18)F]FDG-PET was negative, while CT was positive for the presence of lung metastasis; no viable tumor tissue was found at histological analysis. The remaining patient had a positive [(18)F]FDG-PET and CT study for lung metastasis; this was confirmed at histological analysis. In the same patient, both [(18)F]FDG-PET and CT depicted the presence of a liver lesion. Necrotic lesion regression after treatment was clearly documented with [(18)F]FDG-PET only. CONCLUSIONS Our preliminary results suggest that [(18)F]FDG-PET may be useful for the assessment of metastatic disease in patients with GTTs.

The role of DNA ploidy in postoperative management of stage I endometrial cancer.

BACKGROUND Definition of high-risk stage I endometrial cancer (EC) patients who might benefit from adjuvant therapy (AT) is controversial. Decision is on the basis of traditional prognostic factors. We report our experience in which ploidy has found to play a role in clinical practice since 1999. PATIENTS AND METHODS Two hundred and twenty-two patients with stage I EC with a median follow-up of 4.57 years were studied. After primary surgery, patients are chronologically divided in group A, from 1990 to 1998 (n = 141), receiving AT in IC stage and group B, from 1999 to 2003 (n = 81), receiving AT in case of DNA index >1.2 or stage IC grade 3 with unknown lymph node status. We analyzed prognostic factors, survival and relapse rate of the two groups. RESULTS Since ploidy was introduced as a decision-making factor, only 30.6% (n = 11) of patients with stage IC received AT. Despite this considerable decrease of AT, no tumor-related deaths were reported in the group of patients with diploid IC stage who did not receive AT. Only DNA ploidy and age at diagnosis were independent predictors of overall survival. CONCLUSIONS Our results indicate the important role of ploidy in order to identify high-risk patients who need AT and avoid overtreatment.