Crosstalk between mast cells and pancreatic cancer cells contributes to pancreatic tumor progression.

PURPOSE: To assess the clinical and pathologic significance of mast cell infiltration in human pancreatic cancer and evaluate crosstalk between mast cells and cancer cells in vitro. EXPERIMENTAL DESIGN: Immunohistochemistry for tryptase was done on 53 pancreatic cancer specimens. Mast cell counts were correlated with clinical variables and survival. Serum tryptase activity from patients with cancer was compared with patients with benign pancreatic disease. In vitro, the effect of pancreatic cancer-conditioned medium on mast cell migration was assessed. The effect of conditioned medium from the human mast cell line, LAD-2, on cancer and normal ductal cell proliferation was assessed by thymidine incorporation. Matrigel invasion assays were used to evaluate the effect of mast cell-conditioned medium on cancer cell invasion in the presence and absence of a matrix metalloproteinase inhibitor, GM6001. RESULTS: Mast cell infiltration was significantly increased in pancreatic cancer compared with normal pancreatic tissue (11.4 +/- 6.7 versus 2.0 +/- 1.4, P < 0.001). Increased infiltrating mast cells correlated with higher grade tumors (P < 0.0001) and worse survival. Patients with pancreatic cancer had elevated serum tryptase activity (P < 0.05). In vitro, AsPC1 and PANC-1 cells induced mast cell migration. Mast cell-conditioned medium induced pancreatic cancer cell migration, proliferation, and invasion but had no effect on normal ductal cells. Furthermore, the effect of mast cells on cancer cell invasion was, in large part, matrix metalloproteinase-dependent. CONCLUSIONS: Tumor-infiltrating mast cells are associated with worse prognosis in pancreatic cancer. In vitro, the interaction between mast cells and pancreatic cancer cells promotes tumor growth and invasion.

Detection of M2 macrophages and colony-stimulating factor 1 expression in serous and mucinous ovarian epithelial tumors.

Tumor-associated macrophages (TAM) are known to possess the immunosuppressive M2 macrophage phenotype. They contribute to tumor growth, invasion, and metastasis by producing various mediators. Macrophages, especially M2 polarized macrophages, preferentially express CD163 and CD204, but few studies have investigated macrophage phenotypes in human ovarian tumors. The purpose of the present study was therefore to present results on macrophage differentiation in human ovarian serous and mucinous epithelial tumors. The method focused on immunostaining of paraffin-embedded tumor samples. Almost all macrophages infiltrating tumor tissues expressed CD163 and CD204, indicating the phenotypic shift toward M2 macrophage. The numbers of CD68-positive macrophages as well as of CD163- and CD204-positive macrophages in borderline and malignant tumors were significantly higher than in benign tumors. They correlated well with histological gradient of malignancy. Macrophage colony-stimulating factor (also known as colony-stimulating factor; CSF-1), which is one of the cytokines considered to induce TAM to polarize toward an M2 phenotype, was then evaluated. CSF-1 expression in malignant tumor cells was significantly higher than that in benign tumor cells and correlated with histological malignancy. These results suggest that CSF-1 derived from tumor tissues induces macrophages to shift toward the M2 phenotype, which is considered to promote tumor growth.

[Transvaginal color doppler ultrasonography and CA125 detection in surveillance of ovarian malignant tumors after operation].

BACKGROUND & OBJECTIVE: Although transvaginal ultrasonography color Doppler flow imaging (TV-CDFI) has been widely applied in ultrasonography diagnosis, studies on TV-CDFI combined with a serum tumor marker CA125 detection in the surveillance of ovarian malignant tumors after operation are few. This study was to evaluate TV-CDFI combined with CA125 detection in the surveillance of ovarian malignant tumors after operation. METHODS: Sixty-two cases of ovarian malignant tumors after operation were examined by TV-CDFI. The level of CA125 was measured, and the results were evaluated compared with pathologic findings. The diagnostic sensitivity, specificity, and accuracy were calculated. RESULTS: The diagnostic sensitivity of TV-CDFI or CA125 alone was 84.6% and 65.4%; the specificity was 88.9% and 66.7%; the accuracy was 87.1% and 66.1%. While the diagnostic sensitivity of TV-CDFI combined with CA125 was 92.3%, the specificity was 88.9%, and the accuracy rate rose to 90.3% (P<0.05). CONCLUSION: The diagnostic accuracy for recurrent ovarian malignant tumors is improved by TV-CDFI combined with CA125, which is more helpful in the surveillance of recurrent ovary malignant tumors than TV-CDFI alone.

Características inmunofenotípicas y clínicas del carcinoma renal mucinoso tubular y de células fusiformes / Immunohistochemical profile and clinical features of mucinous tubular and spindle renal cell carcinoma

Introducción: El carcinoma renal mucinoso y tubular de células fusiformes es una neoplasia recientemente incluida en la clasificación de tumores renales. Aún desconocemos aspectos moleculares y de su evolución. Nuestros objetivos son describir características inmunofenotípicas del tumor y aportar casos adicionales para un mejor conocimiento de la neoplasia. Material y métodos: Realizamos inmunohistoquímica con un amplio panel de anticuerpos a 4 tumores (0,89% de carcinomas renales en nuestro centro). Correlacionamos con hallazgos clínicos y tratamos de determinar patrones de inmunotinción. Resultados: Todos los casos marcaron intensa y difusamente para citoqueratina (CK)7. La inmunotinción fue variable para antígeno de membrana epitelial, vimentina, S-100, antígeno Ulex europaeus, CK de alto peso molecular, CK8, CK18 y CK19. Los tumores fueron homogéneamente negativos para antígeno carcinoembriónico, CD15, CD10, CD34, desmina, actina, CK10, CK20 y HMB45. El índice proliferativo (Ki67) fue menor del 1% en todos los casos. La marcación fue similar en células cúbicas y fusiformes. Conclusiones: Este tumor presenta un inmunofenotipo variable de acuerdo con diferentes trabajos y no se conoce con precisión su histogénesis o la célula hacia la cual se diferencian. La similitud en la expresión de marcadores en células cúbicas y fusiformes confirma un mismo origen y diferenciación

Introduction: We present the immunohistochemical (IHC) analyses of a series of four kidney tumors currently classified as mucinous tubular and spindle renal cell carcinoma (WHO), a tumor with uncertain histogenesis and differentiation. Our aims were to determine an immunoprofile and to add clinical and morphological information about this rare renal carcinoma. Material and methods: The four tumors were found between 415 renal carcinomas at our center (0.89%). IHC was carried out with a panel of antibodies in these four cases. Morphogical and clinical information was collected and analyzed. Results: All tumors were well circumscribed and confined to the kidney. Immunoreactivity for cytokeratin (CK)7 was intense and diffuse. Immunostaining was variable for EMA, vimentin, S-100, Ulex europaeus antigen, high-molecular-weight CK, CK 8, CK18, and CK 19. The tumors were homogeneously negative for CEA, CD15, CD10, CD34, desmin, actin, CK10, CK20, and HMB45. Proliferative index (Ki67) was <1% en all cases. The immunoreactivity was similar in cuboidal and spindle cells. Conclusions: The present tumor shows a variable immunophenotype and there is not a precise cell type differentiation. The immunostaining in cuboidal and spindle cells suggests a similar differentiation and histogenesis

Osteoclast-like giant cell tumor in mucinous cystadenocarcinoma of the pancreas: an immunohistochemical and molecular analysis.

Osteoclast-like giant cell tumors (OLGT) are rare neoplasms of the pancreas and mostly associated with ductal adenocarcinomas. In this report, we present the rare case of OLGT associated with mucinous cystadenocarcinoma (MCC). We investigated the expression profile of both tumors by methods of molecular biology and immunohistochemistry. The panel of markers included osteopontin, her2/neu, mismatch repair genes, K-ras, p53, E-cadherin, VEGF-C, and podoplanin. Osteopontin was expressed by the osteoclast-like giant cells but not by the mononuclear tumor cells of the OLGT. We detected an amplification and overexpression of her2/neu in the MCC but not in the OLGT. Although we observed an immunohistochemical expression of hMSH2 and hMLH1 in the OLGT, we were not able to confirm this result by western blot analysis. We also did not find any microsatellite instability (D2S123, BAT26). While mutation of K-ras codon 12 was found in both tumor components, there was wild-type DNA of p53. E-cadherin was expressed in MCC but not in OLGT. VEGF-C was only positive in osteoclast-like giant cells and some of the mononuclear cells of OLGT. The vessel-rich stroma of OLGT did not present any podoplanin-positive lymphatic vessel. The observation of our case and others in the published literature may indicate separating OLGT with undifferentiated carcinoma from OLGT with MCC for the better clinical outcome of the latter.

[Relationship between interleukin-10 level in ascites of patients with primary ovarian epithelial carcinoma and immune defect in peritoneal cavity].

BACKGROUND & OBJECTIVE: As a multifunctional Th2-cytokine, interleukin-10 (IL-10)plays a major role in the immune response. It is well known that IL-10 is an immunosuppressive cytokine, and participates in the development and progression of various tumors. In this study, we investigated the relationship between the IL-10 level in the ascites of the patients with primary ovarian epithelial carcinoma (POEC) and immune defect in the peritoneal cavity. METHODS: The IL-10 levels in serum and ascites of 32 patients with POEC, in culture supernatants of 4 different ovarian carcinoma cell lines and in serum of 10 patients with ovarian epithelial benign tumor and 10 health women (control) were measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: (1) IL-10 level in ascites was significantly higher than that in serum of patients with POEC, (159.78+/-51.20 ng/L vs 12.01+/-4.38 ng/L, P=0.000). IL-10 level in serum of the patients with POEC (12.01+/-4.38 ng/L) was significantly higher than that of the patients with benign tumor (3.79+/-2.40 ng/L, P=0.000) and control (4.45+/-2.69 ng/L, P=0.003). There was no significant difference of IL-10 level in serum between ovarian benign tumor and control (P=0.529). (2) IL-10 level in ascites of the patients with POEC was correlated with FIGO stage but not correlated with histological grade. (3) IL-10 was detectable in culture supernatants of 4 different ovarian cancer cell lines (3ao, SKOV3, CAOV3 and OVCAR). CONCLUSION: High level of IL-10 in ascites of the patients with POEC is probably associated with immune defect in their peritoneal cavity, ovarian cancer cells may promote metastasis in peritoneal cavity by secreting IL-10.

[Borderline ovarian tumors]. / Iaichnikovi tumori s granichna malignenost.

Forty three patients with borderline ovarian tumours were examined from 1987 till 2000. 28 patients (65.1%) were serous tumours and 15 patients (35%) were mucinous. The median age was 42.2 +/- 15.4 years. The follow-up period was 5.4 +/- 3.2 years. Preoperatively Ca125 were 52.5 +/- 50.3 and of Ca19.9-42.4 +/- 46.8 IU. 83% of the patients were in stage I and 17% in stage III. There was no significant difference in the tumour volumes. Ca125 were significantly increased in the serous tumours and Ca19.9 were increased in the mucinous tumours. All the patients were followed up and are in good health or remission with exception of 2 who developed recurrence.

[The role of cytokeratin 7 in the differential diagnosis of primary ovarian carcinoma and metastatic ovarian carcinoma originated from the gastrointestinal tract].

OBJECTIVE: To assess the role of cytokeratin 7 monoclonal antibody in the differential diagnosis of primary ovarian carcinoma and metastatic ovarian carcinoma originated from the gastrointestinal tract. METHODS: Immunohistochemical study using cytokeratin 7 monoclonal antibody and ABC kit. RESULTS: All the 46 cases of primary ovarian carcinoma were CK 7 positive, while in the metastatic ovarian carcinoma of intestinal origin, all cases remained negative for CK7. Half of the 34 cases of metastatic ovarian carcinoma of gastric origin were CK 7 positive. The positive result of CK7 was significantly higher in the primary ovarian carcinoma than in each group of the metastatic ovarian carcinoma (P < 0.001). CONCLUSION: CK 7 is seemed to be a useful antibody in the differential diagnosis of ovarian carcinoma.

[The dose-response of cell proliferation and secretion of CA125 II antigen to follicle stimulating hormone in ovarian cancer cell lines].

OBJECTIVE: To study the dose-response of the proliferation and secretion of CA125 II antigen with follicle stimulating hormone (FSH) in epithelial ovarian cancer cell lines. METHODS: Two different kinds of ovarian cancer cell lines originated from different pathologic types-AO and 3AO were studied. Cancer cells were cultured in medium containing FSH in different concentrations. The proliferation rate of cells was detected by using the 3H-TdR intrusion technique. The accordant titer of CA125 II antigen in the culture medium was measured by enzyme linked immunoadsorbent assay technique. RESULTS: Proliferation rate of cancer cells (AO and 3AO) was increased apparently as increasing in the concentration of FSH in the culture medium. A positive correlation was found between them (P < 0.05). Proliferation rate of AO cells positively correlated with secreting quantity of CA125 II antigen (P < 0.05), however, no such correlation was found in 3AO cells. CONCLUSION: It suggested that FSH can stimulate the proliferation of epithelial ovarian cancer cell at high concentration and there is a accordant dose-response relationship between them. The secreting pattern of the CA125 II antigen varied by the pathological classification of the ovarian cancer.

[Cellular adhesion molecule (CD44) in ovarian tumors].

OBJECTIVE: To investigate a method of detecting adhesion molecule (CD44) contents in ovarian tumors and its clinical significance. METHODS: In 106 patients with ovarian tumors (50 benign and 56 malignant), the adhesion molecule (CD44) contents in peripheral blood lymphocytes (PBL) and in neoplastic tissues were analysed (quantitatively by using flow cytometric-immunological method). RESULT: (1) The CD44 contents in the PBL of the patients with malignant tumor were obviously higher than those of patients with benign tumors (P<0.05) and than those of the control (P < 0.01); (2) Within the group of patients with malignant tumors, the CD44 contents in the neoplastic tissues were higher than those in the PBL (0.01 < P < 0.05); There was no significant difference in CD44 contents among the different histopathological types, whether benign or malignant; (4) The CD44 contents in the PBL decreased gradually after surgery. CONCLUSION: To analyse the CD44 contents in PBL and neoplastic tissues by flow cytometry may be an important method or parameter to differentiate benign ovarian tumors from malignant tumors and to discover recurrence of metastasis of malignant tumors in early stage.

Induction of immunocellular resistance to IL-2-activated lymphocytes within ovarian carcinoma cells.

The development of resistance within ovarian carcinoma cells to activated cytotoxic lymphocytes was the objective of this study. Primary ovarian carcinoma cells were obtained from the ascites of a patient. These cells were cocultured with IL-2-activated autologous tumor-associated lymphocytes (TALs) for 1 week. The resulting selected cells underwent a second coculture for 3 days with IL-2-activated autologous TALs or tumor-infiltrating lymphocytes (TILs). Phenotype analysis of the lymphocytes was performed prior to selection and 4-hr chromium release assays were used to detect resistance induction. Resistance to all effector cells could be demonstrated for the selected cells. However, selected cells maintained in culture demonstrated no difference in cytotoxic susceptibility from unselected cells. The following conclusions were made: (i) rapid immunoselection can occur for ovarian carcinoma in vitro; (ii) the resistance induced is not MHC-restricted; (iii) resistance induced by one type of cytotoxic cell results in general resistance to other types of cell from the same patient; and (iv) this resistance is not maintained during in vitro culture. These results may have direct implications on the future immunotherapy for this condition.

Establishment of OVS1 and OVS2 monoclonal antibodies recognizing human ovarian mucinous cystadenocarcinoma.

Two newly established murine monoclonal antibodies (MAbs), OVS1 and OVS2, to human ovarian mucinous cystadenocarcinoma were further characterized for diagnostic efficacy. The specific SA-1 antigen, purified from the tumor extract was identified as a glycoprotein of 29 kDa. A double determinant biotinstreptavidin alkaline phosphatase immunoassay system, containing OVS1 and OVS2 MAbs was used to determine the SA-1 levels in serum. The OVS1 MAb was used as a first antibody because of its high specificity of 96% while OVS2 MAb, with a lower specificity of 8% but greater sensitivity of 78%, was chosen as a second antibody. Matched sera of 64 healthy controls and 90 patients with definite diagnoses of 25 benign diseases, 14 nonovarian cancer and 51 ovarian cancer, were simultaneously measured together with CA 125 values. At cut-off levels of 220 and 360 units/ml, the SA-1 test showed 63% and 43% positive rates respectively in all types of ovarian cancer, compared to 65% and 57% positive rates for CA 125 at cut-off levels of 35 and 60 units/ml, respectively. Sensitivity for SA-1 at 220 units/ml cut-off level in mucinous ovarian cancer was 75% and increased significantly to 85% when the test was combined with CA 125 at 35 units/ml cut-off level. Furthermore, The combination of both tests significantly increased the positive rates to 86% in all types of early stage ovarian cancer.(ABSTRACT TRUNCATED AT 250 WORDS)

Immunocytochemical determination of EGFR, OV 632 and OC 125 in primary ovarian cancer patients.

An immunocytochemistry study was performed to compare the immunoreactivity of the monoclonal antibodies OC 125, EGFR and OV 632 with smear imprints from 60 ovarian tumours (21 serous cystadenocarcinomas, 12 mucinous cystadenocarcinomas, 9 endometrioid carcinomas, 5 clear cell carcinomas and 5 mixed tumours). Twenty-six patients were premenopausal and 34 postmenopausal. The results showed that 75% of mucinous cystadenocarcinomas were negative for all 3 antigens as were 2 of the 5 (40%) clear cell carcinomas. All other tumours were positive for at least one of the three antigens. OV 632 had an overall sensitivity of 73.3%, EGFR 55% and OC 125 46.6%. Four tumours were OC 125-positive and OV 632-negative. There was no significant difference in positivity of OV 632, OC 125 and EGFR, between pre- and post-menopausal patients (chi 2 = 0.03) or between tumours of stage I and stage III [chi 2 = 0.075 (EGFR) 0.95 (OV 632) and 10.49 (OC 125)]. OV 632 was conducted to be the most sensitive antibody in all types of tumours but OC 125, while less sensitive by itself, increased the overall sensitivity in combination with OV 632 from 73.3 to 80% (not statistically significant).

Monoclonal antibody, Mab 12C3, is a sensitive immunohistochemical marker of early malignant change in epithelial ovarian tumors.

A murine monoclonal antibody (MAb 12C3) that is specific to human ovarian carcinomas was generated by immunizing mice with a human ovarian germinoma cell line (JOHYC-2). The antigen distribution that was defined by MAb 12C3 in normal and malignant human tissues was analyzed by immunohistochemistry on paraffin-embedded and frozen sections. The antibody reacted with 67.7% (21 of 31 cases) of epithelial ovarian carcinomas (6 of 12 cases of serous cystadenocarcinoma, 5 of 7 cases of mucinous cystadenocarcinoma, 7 of 9 cases of clear cell carcinoma, 3 of 3 cases of endometrioid adenocarcinoma), but did not react with any of the benign epithelial ovarian adenomas tested. Partial regions of borderline ovarian malignancies that exhibited marked papillary projection of the lining cells with cellular atypia reacted positively with MAb 12C3 in 14 of 25 cases (56.0%). The histologic features of regional early malignant change corresponded to the expression of the MAb 12C3 epitope in the borderline malignant tumor cells. There was a low frequency of reaction (4.3%) between the antibody and other gynecologic and nongynecologic malignancies (46 cases of 12 tissues). In normal tissues, the antibody reacted positively with only three tissues, including corpora lutein cells, excretory ducts in the submandibular gland, and basal cells of the sebaceous glands. The antigen epitope defined by MAb 12C3 was present on a glycoprotein with a molecular mass of 200 kDa and did not exhibit any cross-reactivity with other well-known tumor markers. These data suggest that MAb 12C3 may be a useful tool for the immunohistologic detection of early malignant changes in epithelial ovarian tumors. In addition, MAb 12C3 also may facilitate a differential diagnosis between benign and borderline malignancies.

[Soluble interleukin-2 receptor level in the sera and ascitic fluids in patients with ovarian cancer].

In 31 patients with ovarian cancer, the sIL-2R level of the sera and ascitic fluids were measured by ELISA, to investigate the inhibitive effect of sIL-2R purified from ascitic fluids on normal lymphocyte transformation, stimulated with phytohemagglutinin (PHA). The results showed that the sera sIL-2R levels in the patients were much higher than those in the normal controls (857 +/- 428kU/L vs 235 +/- 90kU/L, P < 0.001). The sera sIL-2R levels in mucinous cancer were significantly higher than those in serous cancer (988 +/- 539kU/L vs 488 +/- 233kU/L P < 0.01). But no obvious correlation was observed with the histopathological grading, nor with metastasis. Higher levels of sIL-2R were also observed in the ascitic. The normal lymphocyte transformation stimulated with PHA was significantly inhibited by high sIL-2R purified from the ascitic fluids.

Epithelial antigens in carcinomas of the ovaries. Relation to histological classification.

In an attempt to assess and improve the histological classification of ovarian tumors the value of immunohistochemical techniques has been examined in 50 ovarian tumors. A panel of six immunohistochemical markers (two cytokeratins, EP4, EMA, CEA, and vimentin) seems to have no additional value in differential diagnosis and typing of ovarian tumors.

[Sharing antigenic determinant of colon carcinoma between serous and mucinous tumors of the ovary].

Expression of colon-ovarian tumor antigen (COTA) in serous and mucinous tumors of the ovary was determined immunohistochemically, using monoclonal antibody SC13A against colon carcinoma antigen as a probe. In 67 serous tumor of the ovary, the frequency of expression was 90.0% in the cystadenocarcinomas, 71.4% in the borderline cystadenomas, and only 8.0% in the cystadenomas. In 44 mucinous tumors of the ovary, the SC13A marker was expressed in 87.5% of the malignant tumors, 80.0% of the borderline tumors, and 25.8% of the benign cystadenomas. And in 20 samples of normal ovarian tissue a negative immunostaining for the SC13A was found. These findings showed that the serous and the mucinous tumors share the same antigenic determinant of colon cancer, and the percentage expression of which is significantly higher in the malignant than in the benign and the borderline tumors, as well as in the normal ovarian tissues (P < 0.005). The marker might be valuable for further studies of these tumors.

Immunomodulation in patients with epithelial ovarian cancer after adoptive transfer of tumor-infiltrating lymphocytes.

The immunomodulation determined by natural killer cell activity, delayed-type hypersensitivity to purified protein derivative and phytohemagglutin, and phenotypic changes of peripheral blood lymphocytes was characterized in 12 patients with epithelial ovarian cancer who received adoptive transfer of tumor-infiltrating lymphocytes (TILs) after cisplatin-containing chemotherapy (TIL group). As a control, 10 patients with epithelial ovarian cancer who did not receive infusions of TIL were also examined in the same fashion. In the TIL group, peripheral blood lymphocytes showed increased percentages of cells bearing the CD8 antigen, in contrast to stable percentages of CD4 antigen-bearing cells, resulting in a decreased ratio of CD4+ to CD8+ cells. The percentages of CD16 and CD56 antigen-bearing cells also increased in proportion to augmentation of natural killer cell activity against K562 cells. Additionally, with regard to cell-mediated immunity determined by delayed-type hypersensitivity to phytohemagglutin and purified protein derivative, significantly and slightly enlarged erythema was observed 2 and 8 weeks, respectively, after the injection of TILs (phytohemagglutin, P < 0.05; purified protein derivative, not statistically significant). The control group showed no major changes in any of the immunological markers. These results suggest the possibility that the adoptive transfer of TILs induces immunoactivation of cellular immunity and enhances natural killer activity in patients with epithelial ovarian cancer.