Mauro Modesti

Immunity to Extracellular Matrix Antigens is Associated with Ultrastructural Alterations of the Stroma and Stratified Epithelium Basement Membrane in the Skin of Hashimoto's Thyroiditis Patients:

Employing purified extracellular matrix (ECM) proteins, i.e. type I, III, IV and V collagens (CI, CHI, CIV, CV), laminin (LM) and fibronectin (FN), as antigen sources we detected autoantibodies to conformational and/or denatured ECM antigens among 34 of 50 sera obtained from Hashimotos thyroiditis (HT) patients and 6 of 51 control sera obtained from non-autoimmune thyroid disease patients and healthy donors (HT sera vs. control sera p=4×10−9). Reactivity to conformational antigens, mostly due to autoantibodies of the IgG isotype, was observed in 30/50 HT sera and in 6/51 control sera (p=3.5×10−7) and was not always concomitant with that to linear antigens, found in 23/50 HT and in 6/51 control sera (p=1.6×10−4). Ultrastructural analysis of skin biopsies obtained from 18 HT patients without symptomatic cutaneous diseases revealed defects of the stratified squamous epithelium basement membrane in 11/18, alterations of the stroma in 13/18 and both basement membrane and stromal defects in 9/18. Interestingly, 13/13 (p=0.012) and 9/11 (p=0.012) patients with stromal and basement membrane defects respectively, exhibited serum antibodies to at least one ECM antigen involved in the organization of the altered tissue compartment. Lastly, 10/18 skin biopsies presented immunoglobulin (Ig) and/or complement (C3) deposits along the cutaneous basement membrane zone (BMZ) or in the papillary dermis and 9/10 sera from the same patients simultaneously showed antibodies to at least one ECM antigen involved in the organization of these two skin compartments. Besides, 8/11 HT patients with basement membrane defects exhibited Ig or C3 deposits along the BMZ. Our findings suggest that autoantibodies to ECM molecules might contribute to the development of asymptomatic extra-thyroid skin diseases in HT patients.

Local delivery of recombinant vaccinia virus encoding for neu counteracts growth of mammary tumors more efficiently than systemic delivery in neu transgenic mice

Recombinant vaccinia virus has been widely employed as a cancer vaccine in several clinical trials. In this study we explored, employing BALB/c mice transgenic for the rat neu oncogene, the ability of the recombinant vaccinia virus neu (rV-neuT) vaccine to inhibit growth of neu+ mammary carcinomas and whether the efficacy of vaccination was dependent on: (a) carcinogenesis stage at which the vaccination was initiated; (b) number of vaccinations and (c) route of delivery (systemic vs. local). BALB-neuT mice were vaccinated one, two and three times by subcutaneous (s.c.) and intramammary gland (im.g.) injection with rV-neuT or V-wt (wild-type vaccinia virus) starting at the stage in which mouse mammary gland displays atypical hyperplasia, carcinoma in situ or invasive carcinoma. We demonstrated that vaccination using rV-neuT was more effective when started at an earlier stage of mammary carcinogenesis and after three vaccinations. The im.g. vaccination was more effective than the s.c. vaccination in inhibiting mammary carcinogenesis, eliciting anti-Neu antibodies, increasing anti-Neu IgG2a/G3 isotypes and inducing antibodies able to trigger mammary tumor cells apoptosis and antibody-dependent cellular cytotoxicity. The better protective ability of rV-neuT im.g. vaccination was associated with its capacity to induce a superior degree of in vivo mammary cancer cells apoptosis. Our research suggests that intratumoral vaccination using recombinant vaccinia virus could be employed to increase the activity of a genetic cancer vaccine. This study may have important implications for the design of cancer vaccine protocols for the treatment of breast cancer and of accessible tumors using recombinant vaccinia virus.

Metastasis to the Pancreas from Breast Cancer: Difficulties in Diagnosis and Controversies in Treatment

Background: Metastasis to the pancreas originating from malignant tumours is a rare event and, in the literature, we have found only 11 reported cases of solitary pancreatic metastases originating from breast cancer. Case Report: We report a case of a 51-year-old woman with primary breast cancer who developed obstructive jaundice and epigastric pain after 2 years without any symptoms. The pancreatic mass revealed by computed tomography (CT) scan and magnetic resonance imaging (MRI) was not recognised as a metastasis from breast cancer and the patient underwent cephalic pancreaticoduodenectomy. Conclusions: We discuss all aspects of the case management, stressing the importance of a careful evaluation of the clinical history and the primary cancer features and the usefulness of a multi-disciplinary approach. These aspects are of main importance for a correct diagnostic process and an appropriate therapeutic choice when a pancreatic lesion develops in a patient with prior neoplasm.

Ewing's sarcoma lines synthesize laminin and fibronectin

Immunoelectron microscopy was employed to detect laminin and fibronectin cell surface expression on five Ewings sarcoma lines plus a normal fibroblast line as control. Monospecific antibodies to both glycoproteins were detected on tumour cell and fibroblast layers with colloidal gold - protein A conjugates. All five tumour lines were positive for fibronectin and/or laminin, whereas the fibroblast line expressed fibronectin only, as expected. Fibronectin displayed a dense granular pattern, typically in the cell-cell and cell-matrix adhesion areas; laminin displayed a punctate pattern.3H-leucine metabolical labelling was also used to demonstrate laminin and fibronectin synthesis. The labelled proteins released in the culture media were separated by molecular weight on SDS-PAGE and identified by immunoprecipitation with the monospecific antibodies. The results substantiated the immunoelectron microscopy data. These findings indicate that Ewings sarcoma lines produce a complex extracellular matrix including fibronectin and laminin, in addition to the collagens described by other workers. Histogenetic classification of this tumour in terms of extracellular matrix proteins synthesis is thus more difficult than has been supposed. The same complexity must also be borne in mind when using the matrix components as an aid to Ewings sarcoma differentiation from other childhood tumours.

EXTRACELLULAR MATRIX REMODELLING IN A MURINE MAMMARY ADENOCARCINOMA TRANSFECTED WITH THE INTERFERON‐ALPHA1 GENE

The rejection of interferon alpha1 gene‐transfected mammary adenocarcinoma cells (TSA‐IFNα) injected into syngeneic BALB/c mice was accompanied by an unusual stromal reaction and marked CD8‐positive T‐lymphocyte involvement. To investigate the biological background of this reaction, the possibility was evaluated that an interaction between TSA‐IFNα and stromal cells might remodel the extracellular matrix (EM). When fibroblasts were co‐cultured with TSA‐IFNα or treated with exogenous IFNα, there was no change in their replication rate or collagen synthesis. By contrast, their fibronectin (FN) production and release were increased, resulting in enhanced fibroblast chemotaxis. These findings were mirrored by increased FN staining in the peritumoural and tumoural areas in vivo. IFNα thus determines increased FN production and hence massive local recruitment and activation of fibroblasts, with a modification of the EM. The several activities of IFNα should thus be considered prior to its employment in clinical trials.