[Show abstract][Hide abstract] ABSTRACT: The neonatal Fc receptor (FcRn) was demonstrated to play a role both in the recycling and thus the protection of immunoglobulin G (IgG) from catabolism and in the maternal-fetal transfer of IgG. The expression of this particular receptor was evidenced in a variety of cell types, but the endothelial cell was considered the main cell able to perform both recycling and IgG catabolism. Based on preliminary data obtained in adult human mammary glands and skin, this study focused on a number of neonatal human tissues, targeting FcRn expression mainly in epithelial versus endothelial cells. Our results demonstrate that in most of the investigated tissues, the neonatal Fc receptor is not detectable in the endothelial cells lining the capillaries, whereas most epithelial cells are positive. We could also observe the receptor's expression in most macrophages, smooth muscle cells, and neurons. Taken together, these data suggest that the main sites of IgG catabolism might in fact be other than endothelial cells in human neonates.
Human immunology 12/2011; 72(12):1176-87. DOI:10.1016/j.humimm.2011.08.020 · 2.14 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Endometrioid endometrial carcinoma developed from endometrial hyperplasia is associated with anomalies of proliferation, apoptosis, and matrix metalloproteinase (MMP) expression. Our study was designed to investigate steroid receptor (ER, PR) expression and its correlation with proliferative activity (PCNA), apoptosis (Fas, FasL, Bcl-2, Bax, and p53), gelatinases (MMP-2 and MMP-9) and their tissue specific inhibitor (TIMP-1 and TIMP-2) immunoexpression in endometrial carcinogenesis. A total of 38 cases were investigated, 10 non-neoplastic, 11 hyperplastic, and 17 carcinomatous endometria. Immunolabeling showed a higher expression of steroid receptors in hyperplasia and carcinoma than in non-neoplastic endometria and an ER/PR imbalance in carcinoma. The epithelial component of endometrial carcinomas had the highest proliferative index. Bcl-2 had a stronger expression in hyperplasia and carcinoma compared to non-neoplastic endometria and stromal tissue. The Bcl-2/Bax ratio was lower in endometrial carcinoma. Fas and FasL expression was stronger in hyperplasia and furthermore in carcinoma. p53 expression was progressively stronger along the sequence non-neoplastic endometrial to hyperplasia-carcinoma. Both types of investigated MMPs showed an increased expression in neoplastic endometria reaching a maximum level in carcinomas. MMP-9 immunostaining could be correlated to myometrial invasion. TIMP-1 decreased and TIMP-2 increased in expression from non-neoplastic endometria to hyperplastic and carcinomatous endometrial, respectively. Our study demonstrates that coordinated anomalies of steroid receptors, apoptosis and invasiveness factors are already present in hyperplasia as cumulative steps along the way to malignant transformation and that a complex MMP-2, MMP-9, TIMP-2/TIMP-1 imbalance seems to be responsible for the endometrial proliferation.
Annals of anatomy = Anatomischer Anzeiger: official organ of the Anatomische Gesellschaft 02/2011; 193(1):43-55. DOI:10.1016/j.aanat.2010.09.009 · 1.48 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Analysis of protein phosphorylation within intracellular signaling pathways may clarify the functional status and biological responses of cells to various stimuli, reflecting an enhanced enzymatic activity. Phos-pho-specific antibody (p-Ab)-based detection methods that have been used to date, including flow cytometry, re-quire extensive optimization for the achievement of accuracy and rational specificity. Objective. We sought to op-timize a phospho-specific flow cytometry detection method, based on the premise that fixation is a critical step of such protocols. Materials and methods. We compared two different methods for phospho-detection in phorbol myristate acetate (PMA)-activated peripheral blood mononuclear cells (PBMCs) and assessed how they dis-tinctly impacted on cell viability, surface staining intensity and phospho-detection competence, aiming to develop an optimized protocol with balanced efficiency in surface and intracellular phospho-labeling. PMA-activated PBMCs were used for testing the detection efficiency of five p-Abs: pERK(T202/Y204), pp38MAPK(T180/Y182), pAKT(T308), pSTAT-3(S727), pSTAT-1(Y701) by flow cytometry. Results. 1. Fixation prior to surface staining led to an improved signal-to-noise ratio for all p-Abs evaluated; 2. cell suspension integrity was not affected by the fixation method used; 3. reduced efficiency of surface marker detection levels remained within acceptable ranges; 4. one freeze-thaw cycle did not significantly impair the cell suspension integrity or staining efficiency. Conclusion. These results provide a practical approach for phospho-detection in heterogeneous tissue samples. The clinical relevance of our technical approach appears substantial, as frequent dysfunctions within signaling pathways are linked to various diseases and consequently require proper identification methodology.
Revista Romana de Medicina de Laborator 12/2010; 18(4):55-65. · 0.24 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: A 57-year-old woman, with left choroidal melanoma treated by laser photocoagulation and a history of repeated vitrectomies, checked for left eye acute pain and foreign body-like sensation, symptoms that occurred after three years since the primary tumor treatment. The left eyeball was enucleated and the tissues were investigated by immunohistochemistry for markers associated with cell differentiation, proliferation and adhesion, cell cycle regulation, apoptosis control, vascularization, invasiveness and local immune response. We identified, in fact, two independent tumors, with different localization and sharing some common features, markers of a highly aggressive potential: loss of cell differentiation markers and cell cycle regulators, ability to avoid death by suppressing Fas antigen expression and important invasive capacity by down regulation of E-cadherin expression. However, only in the posterior tumor, we found cells with high proliferation rate, Fas ligand molecule expression and MMP-9 secretion, acquisitions associated with a much more aggressive behavior. These particular phenotypes allowed the posterior cells to grow and to invade the surrounding tissues more rapidly than the anterior ones, leading to the development of a large size tumoral mass, responsible for the clinical symptoms. Photocoagulation, by destroying the tissues, makes impossible the evaluation of the primary tumor's biological features, important for the tumor evolution. The absence of these data stresses the importance of patient monitoring, eventually addressing a panel of soluble markers associated with recurrence or metastasis development.
Romanian journal of morphology and embryology = Revue roumaine de morphologie et embryologie 01/2010; 51(1):187-93. · 0.66 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: We have identified by immunohistochemistry/ immunocytochemistry the expression of bcl-2 molecule in 55 primary breast carcinomas and in 30 corresponding axillary lymph nodes metastases, together with a set of molecules known as prognostic factors: estrogen receptors, progesterone receptors, and p53 protein. Our results demonstrated a significant correlation (p < 0.05) between bcl-2 and hormonal receptors expression in tumors, but not in axillary metastases (p < 0.1), a significant inverse correlation between bcl-2 and p53 expression in primary tumors (p < 0.02), but a significant direct correlation in axillary metastases (p < 0.02). The bcl-2+/p53- phenotype, associated with normal breast epithelium, is present in 79.17% primary tumors, but only in 15.38% axillary lymph nodes metastases. A larger number of lymph nodes metastases expressed a bcl-2+/ p53+ more aggressive phenotype compared with primary tumors (58.82% versus 48.39%). This shows that changes in the expression of bcl-2, p53, estrogen and progesterone receptors can lead to an increased cellular aggressiveness and thus to an increased tumoral invasive and metastasizing potential.
[Show abstract][Hide abstract] ABSTRACT: We have investigated the cellular and serum CK18 in 26 non-treated primary ductal invasive breast carcinomas. The soluble CK18 (TPS) was detected by chemiluminescent assay, and the cellular CK18 and PCNA expression by immunocytochemistry. Flow-cytometry was used to estimate the amount of DNA in malignant cells. There was a significant correlation between soluble CK18 and the pre-menopausal status (p < 0.05), characterized in our group by a PCNA estimated low proliferation index. We have also found a significant correlation between soluble CK18 and the DNA index (p < 0.01). The intracellular CK18 has correlated with the PCNA expression (p < 0.05), while no correlation could be found between cellular and serum CK18. The values of soluble CK18 may offer information about the treatment-induced cell death, if monitored, while isolated measurements should be interpreted cautiously. Elevated levels of serum CK18 in non-treated carcinomas may rather reflect a high tumor turn-over or perhaps a more intensive tumor cell killing.
[Show abstract][Hide abstract] ABSTRACT: The etiology and pathogenesis of acne vulgaris are not yet completely understood. Therefore we have investigated 5 patients with different clinical forms of disease, including the rare form of acne fulminans. Taking into consideration the four factors that are currently incriminated in the development of acne, sebaceous hypersecretion, hyperkeratosis of the pilosebaceous infundibulum, bacterial colonisation and perifollicular inflammation, we have focused our study on a set of cells involved in the chronic inflammatory process. We have evidenced by immunohistochemistry methods, using appropriate monoclonal antibodies, the presence of T lymphocytes and macrophages, while the B cells could be evidenced only in the severe forms. We were also interested to investigate the occurrence of new capillary formation, as an accompanying phenomenon of the inflammatory process. The presence and histological distribution of these cells highly supports the hypothesis that the mechanisms underlying the development of acne vulgaris belong to the Delayed Type Hypersensitivity.
[Show abstract][Hide abstract] ABSTRACT: The major histocompatibility complex (MHC) class I related neonatal Fc receptor (FcRn) plays multiple roles, being involved in transporting immunoglobulin G (IgG) and protecting this antibody class from catabolism. The presence of this receptor was previously demonstrated in the lactating murine mammary gland. In the current study we have investigated FcRn expression in various histologic types of human breast carcinoma and lymph node metastases. We used immunohistochemical methods to demonstrate the presence of FcRn in epithelial cells, whereas this Fc receptor could not be detected in mammary gland endothelial cells. The presence of the receptor was also found in the metastasizing epithelial cells within the lymph nodes, and this provides a useful marker for their identification.
Human Immunology 01/2004; 64(12):1152-9. DOI:10.1016/j.humimm.2003.08.025 · 2.14 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Fas (CD95/APO-1) and its natural ligand, FasL, are molecules expressed on cellular membranes, being involved in the induction of programmed cells death or apoptosis. Recently, it has been shown that malignant cells originating from solid tumors tend to inhibit the expression of Fas, as an escape mechanism from the immune cells' attack and to express FasL, as a counterstrike mechanism against the immune effector cells. The purpose of this study was to investigate, by immunohistochemistry, the presence of Fas and FasL in 15 breast carcinomas and to establish possible associations between the expression of these molecules and the histological type and grading of the tumors. Our results showed that 7 breast tumors have lost the expression of Fas and 11 tumors were positive for the Fas-ligand expression, important arguments for the mechanisms of immune escape and tolerance induction. Furthermore, 7 of the 11 FasL+ tumors were poorly differentiated invasive ductal carcinomas, suggesting a possible association between FasL expression and tumor aggressivity.
[Show abstract][Hide abstract] ABSTRACT: Breast tumors growth is regulated by female sex steroid hormones. The level of the estrogen and progesterone receptors (ER and PR) expression by the malignant cells is important for the evaluation of the tumor prognostic and the benefit of a hormonal therapy. The aim of our study was to identify the expression of estrogen and progesterone receptors in primary breast tumors and in the corresponding axillary lymph nodes metastases, in 24 cases. The results showed that more than 30% of poorly differentiated breast carcinomas lost their expression of hormone receptors from the primary tumors to axillary metastases, an event which can be associated with an aggressive tumoral behaviour and resistance to hormonal therapy.
[Show abstract][Hide abstract] ABSTRACT: Oncogenes, the abnormal forms of proto-oncogenes, were shown to be involved in malignant transformation and in tumor progression. c-erbB2/HER2/neu is member of EGFR family and encodes the p185 protein, which functions as a tyrosine-kinase. Gene amplification and/or p185 overexpression were reported to be associated with poor prognostic in cancer. Our purpose was to investigate p185 immunohistochemical expression in breast carcinomas and in the corresponding axillary lymph nodes metastases and to identify possible correlation between p185 and other factors of poor prognostic, such as loss of hormonal receptors expression. In our study, 40.91% of cases were erbB-2 positive, p185 expression being maintained from the primary tumors to axillary metastases and associated with positive nodal status and with the absence of hormonal receptors expression (p < 0.05). These findings support the hypothesis the c-erbB2 is an advantageous acquisition for the aggressive behavior of the tumor cell and for its ability to invade and metastasize.
[Show abstract][Hide abstract] ABSTRACT: Malignant transformation is the result of genetic events, translated into sequential acquisitions of multiple abnormalities in the control of cellular growth and cell cycle regulation. We determined the expression of the estrogen receptor (ER), progesterone receptor (PR), c-erbB-2 and p53 gene products in a patient with mixed (ductal and lobular) invasive breast carcinoma bearing different coexisting lesions. The purpose of the study was to establish a possible correlation between the expression pattern for these molecules and the histological appearance of the breast tumor. Our results showed no positivity for ER. PR expression was restricted to normal epithelium, simple hyperplasia and in situ carcinoma. c-erbB-2 was detected in all lesions with the exception of normal epithelium and immunostaining for p53 was found positive only in in situ and invasive carcinoma. These findings support the hypothesis of tumorigenesis as a multistep process and as a sum of changes, each representing an advantageous acquisition for the malignant cells' behavior. The loss of hormone receptors' expression occurred as an early event in this case, while the p53 mutations were found only in more advanced neoplastic lesions.
Journal of B.U.ON.: official journal of the Balkan Union of Oncology 01/2003; 8(3):281-4. · 0.74 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Purpose: This study was designed to detect the presence of the p27(Kip1) protein expression in breast carcinomas and corresponding axillary lymph node metastases, as well as any potential correlation of p27(Kip1) with factors of tumor progression and prognosis (estrogen receptors-ER, progesterone receptors-PR, c-erbB-2, p53). Patients and methods: 44 primary breast cancers and positive axillary lymph nodes from 24 cases were examined by immunohistochemistry. Results: 19 out of 44 (43.18%) tumor specimens expressed the p27 protein, whereas 25 (56.82%) specimens expressed it only at low levels or were negative. Absence of p27 correlated significantly (p < 0.05) with a negative status for ER and PR receptors. The presence of p27 protein in primary tumors was always associated with positive expression in the corresponding nodal metastases. Conclusion: Our results indicate a correlation between lack or low levels of p27 protein expression and the absence of hormone receptors. The p27 phenotype is preserved from the primary breast tumors to the corresponding axillary lymph node metastases.
Journal of B.U.ON.: official journal of the Balkan Union of Oncology 01/2002; 7(2):117-20. · 0.74 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: During normal pregnancy, the maternal immune response achieves a split tolerance state towards the fetal allograft, expressed by immunophenotypic profile. This may be impaired in the preterm birth. Our study aimed to analyze the immunophenotypic profiles of the peripheral and retroplacental blood in women with normal pregnancy and preterm birth. Blood leukocyte populations were assessed by flowcytometry in nine normal at term pregnancies, five preterm births and eight nonpregnant controls. Our results showed in normal pregnant women as compared to controls, a significant decreased number of lymphocytes, monocytes and B cells, while granulocytes, T-lymphocytes and the T CD8+ subsets were elevated. The CD4/CD8 ratio was significantly decreased, especially in the retroplacental blood. The preterm birth group demonstrated a significantly elevated number of lymphocytes and B cells, lower T CD8+ cells and an increased CD4/CD8 ratio, in comparison with normal pregnancies group. CONCLUSION: Normal pregnancy determines a suppressive immunophenotypic profile that is impaired in the preterm birth suggesting its immunological involvement.
[Show abstract][Hide abstract] ABSTRACT: The lymph nodes status is one of the most important prognostic factors in breast cancer. The routine hematoxilin-eosin staining is efficient for the metastases detection only when there is a large number of tumor cells, while a small number of metastatic cells can easily remain undetectable. For those situations, the immunohistochemistry for cytokeratins, markers of the epithelial cells, is a very sensitive method. We have investigated the cytokeratin 8 expression in 10 primary breast carcinomas and in the corresponding axillary lymph nodes, comparing with hematoxilin-eosin. The routine examination has detected axillary lymph nodes metastases in six cases, confirmed by the immunohistochemistry for cytokeratin 8. Four cases were diagnosed as negative for the axillary lymph nodes metastases by the hematoxilin-eosin staining. In all those four cases, immunohistochemistry for cytokeratin 8 has detected a small number of tumor cells, either spread in the lymph nodes tissues, either confluent as small islets.
[Show abstract][Hide abstract] ABSTRACT: The NK cells are large granular lymphocytes that are found as approximately 2-15% of the global peripheral lymphocytes. As endometrial granulocytes, they represent 70% of the decidual leukocytes during the first trimester of pregnancy, with a progressive decreasing until term. The complex actions of these numerous and active cells, consist in: 1) placental regulation and control, based on the receptors for the HLA--G and HL--C antigens, that are trophoblast specific and act as universal inhibitors of the NK cells; 2) secretion of a large range of cytokines as IL-1, TNF alpha, IL-8, TGF, IFN gamma, GM-CSF, CSF1 which regulate the trophoblastic proliferation, differentiation and invasion; 3) removal of abnormal trophoblastic cells, which may appear in such a proliferating cell population; 4) infectious protection of the placental unit, due to their unspecific cytotoxic effect. In conclusion, the NK lymphocytes have important implications in the formation of the decidua, as the maternal side of the fetomaternal interface and they are deeply involved in the evolution of normal pregnancy concerning the nutritional, hormonal and immunologic aspects.
[Show abstract][Hide abstract] ABSTRACT: Angiogenesis represents an essential event required by tumors to support their growth. The aim of our study was to investigate the presence of neovascularization in 44 primary breast carcinomas and in 24 axillary lymph nodes metastases and to establish a possible correlation between the presence of tumor angiogenesis, some clinical and pathological features of the cases and the expression of p53, an important cell cycle regulator. To identify the new blood vessels, we used immunohistochemistry for von Willebrand factor, a marker of the endothelial cells. The results showed that 77.27% of the primary breast carcinomas and 75% of the lymph nodes metastases are positive for von Willebrand factor and this positivity is significantly correlated (p < 0.05) with the expression of p53, supporting the idea that angiogenesis is a marker for tumor aggressiveness and p53 could be involved in this process.
[Show abstract][Hide abstract] ABSTRACT: Adaptive immune responses are not initiated at the site where a pathogen first establishes a focus of infection. They occur in the organized peripheral lymphoid tissues, to which the pathogen or its products are transported, trapped, captured by specialized cells, called antigen-presenting cells (APC), which process and present the antigen to T lymphocytes. Activation of naive T cells requires two signals: the first signal is represented by the specific recognition of a foreign peptide fragment bound to a self MHC molecule, but this is not enough. The second signal, called co-stimulatory signal is represented by other molecules, expressed on the membrane or secreted by the APCs for which T cells express specific ligands. Binding of antigen to the T-cell receptor initiates a series of biochemical changes within the T cell, involving a large number of molecules.