Comparative immunomorphology of testicular Sertoli and sertoliform tumors.

Sertoli cell (SC) and sertoliform tumors of the testis are very uncommon; for this reason their differential diagnosis and classification can be challenging. We applied an extensive immunophenotypic panel that included androgenic hormones, enzymes and receptors, neuroendocrine, lineage and genitourinary markers to a series of these lesions to determine if and which immunostains can aid in their diagnostic workup. Study cases included: 2 androgen insensitivity syndrome-associated SC adenomas, 3 SC tumors (SCT) not otherwise specified (SCT-NOS), 3 sclerosing SCT, 2 large cell calcifying SCT, 1 SCT with heterologous sarcomatous elements, 1 malignant SCT, and 1 sertoliform rete testis adenoma (sertoliform RTA). We found that SCT-NOS and variants with sclerosis showed a phenotype akin to atrophic seminiferous tubules characterized by gain of expression of pankeratin, calretinin, CD56, which are negative in normal SC. Distinctive phenotypes were identified in: sclerosing SCT: androgen receptors (AR) + (strong)/PAX2/PAX8+ (subset)/S100+/inhibin-; large cell calcifying SCT: calretinin+ (strong)/S100+/AR-; sertoliform RTA: PAX2/PAX8+/pankeratin+/inhibin-. Androgenic hormones and enzymes did not show diagnostic utility. A panel of calretinin, inhibin, pankeratin, S100, PAX2/PAX8, and AR consistently allowed distinction between variants of Sertoli and sertoliform tumors.

[Cystadenoma of the liver with high levels of ACE and CA 19-9 in the cyst]. / Cystadénome hépatique avec taux d'ACE et de CA 19-9 intrakystiques élevés.

The development of a cystadenocarcinoma from previously benign cystadenoma is controversial. Neither clinical nor biological factors have been described to explain this transformation. High levels of serum and cystic CEA and CA 19-9 seem to help in the diagnosis of cystadenoma but not cystadenocarcinoma. Definitive histological evaluation is the only means to discriminate malignant from benign cysts. We report a case of cystadenoma of the liver with very high cystic levels and normal serum levels of CEA and CA 19-9.

Diagnostic value of CA 72-4 and carcinoembryonic antigen determination in the fluid of pancreatic cystic lesions.

OBJECTIVE: Mucinous cystic tumours of the pancreas need to be distinguished from other cystic lesions because of their malignant potential. The aim of this study was to assess prospectively the reliability of CA 72-4 and carcinoembryonic antigen analysis in the fluid of cystic lesions of the pancreas obtained by fine-needle aspiration for pathological diagnosis. METHODS: CA 72-4 and carcinoembryonic antigen were measured in cyst fluid obtained preoperatively by fine-needle aspiration. The 91 lesions consisted of 16 serous cystadenomas, 16 mucinous cystadenomas, 14 cystadenocarcinomas and 45 pancreatic pseudocysts complicating well documented chronic pancreatitis. RESULTS: A CA 72-4 level of >40 U/ml had a 63% sensitivity and 98% specificity for distinguishing mucinous cystadenomas and cystadenocarcinomas from serous cystadenomas and pseudocysts. A carcinoembryonic antigen level of >400 ng/ml had a 57% sensitivity and a 100% specificity for distinguishing mucinous tumours and cystadenocarcinomas from pseudocysts. A carcinoembryonic antigen level of <4 ng/ml had a 100% sensitivity and a 93% specificity for distinguishing serous cystadenomas from mucinous cystadenomas, cystadenocarcinomas and pseudocysts. CONCLUSION: Combined measurement of CA 72-4 and carcinoembryonic antigen may be used to distinguish accurately mucinous cystadenomas and cystadenocarcinomas from serous cystadenomas and pseudocysts.

Expression of cadherins and CD44 isoforms in ovarian endometrial cysts.

We evaluated the immunohistochemical expression of cadherins and CD44 variants in 20 endometriomas, 20 cystadenomas, 20 borderline ovarian tumours as well as 20 ovarian carcinomas, and the serological and cystic fluid concentrations of soluble E-cadherin and soluble CD44 standard (sCD44sdt) in 20 endometriomas, 20 cystadenomas, six borderline and 11 carcinomas of the ovary. In endometriomas, immunostaining of E- and N-cadherin was negative (20 and 30% respectively). CD44 H, v3 and v6 immunostaining were detected in 63, 10 and 40% respectively. A difference in immunostaining for E-cadherin was found between endometriomas and cystadenomas (P < 0.001) and for N-cadherin between endometriomas and carcinomas (P < 0.001). A difference in CD44H immunostaining was observed between endometriomas and cystadenomas (P < 0.035) but not with borderline ovarian tumours and carcinomas. No difference in serum concentrations of soluble E-cadherins and CD44 standard was found between the four groups of tumours. Cystic fluid concentrations of E-cadherin were lower in endometriomas than in borderline tumours and ovarian carcinomas (P < 0.001). High concentrations of soluble CD44 standard cystic fluid were found in endometriomas than in other ovarian cysts. Endometriomas and borderline tumours share alterations of cadherins and CD44 isoforms which may help in the understanding of the aggressive and invasive potentials of endometriotic cells.

Thrombin-antithrombin III and D-dimer plasma levels in patients with benign or malignant ovarian tumours.

In 50 patients with benign ovarian tumours, 39 malignant ovarian carcinoma patients and 39 age-matched healthy women, plasma levels of thrombin-antithrombin III complex and D-dimer were determined as well as CA 125. The coagulation activation marker thrombin-antithrombin III complex and D-dimer levels were elevated in the malignant group compared to the benign and control groups. The results suggest that coagulation and fibrinolysis must play a prominent role in ovarian cancer. Moreover, D-dimer and thrombin-antithrombin III were equally useful as CA 125 for the discrimination of patients with benign or malignant ovarian tumours as evidenced by receiver operating and likelihood ratio calculations.

Endocrine cells in hepatobiliary cystadenomas and cystadenocarcinomas.

We investigated the distribution of endocrine cells in hepatobiliary cystadenoma (n = 5, two associated with mesenchymal stroma) and cystadenocarcinoma (n = 3) immunohistochemically. In normal livers (n = 20) and livers affected by hepatolithiasis (n = 15) used as controls, endocrine cells revealed by chromogranin immunostaining were located exclusively in normal or proliferating intrahepatic peribiliary glands. In the eight cases of hepatobiliary cystadenoma and cystadenocarcinoma, endocrine cells were present in four cases (50%) (1 cystadenoma, 1 cystadenoma with mesenchymal stroma, and 2 cystadenocarcinomas). Endocrine cells tended to be located beneath and among the columnar epithelial cells. Intrahepatic peribiliary glands were located in the vicinity of cystadenoma or cystadenocarcinoma in six (75%) of the eight cases, and they frequently showed cystic dilatation and contained endocrine cells. Intrahepatic peribiliary glands were located in the vicinity of the endocrine cells in all cystadenomas and cystadenocarcinomas that were positive for endocrine cells. These data show that about 50% of hepatobiliary cystadenomas and cystadenocarcinomas contain endocrine cells and suggest that hepatobiliary cystadenoma and cystadenocarcinoma may originate from intrahepatic peribiliary glands.

Ki67 antigen immunostaining (MIB 1 monoclonal antibody) in serous ovarian tumors: index of proliferative activity with prognostic significance.

The purpose of this study was to evaluate the biological significance of Ki67 antigen expression in serous ovarian tumors, through the analysis of MIB 1 monoclonal antibody reactivity in cystoadenomas, borderline tumors, and invasive cystoadenocarcinomas; the correlation between this index of cell proliferation and clinicopathologic parameters (FIGO stage and grade, and disease-free survival) was also investigated in invasive cystoadenocarcinomas. Fifty-four patients with serous ovarian tumors, treated at the Institute of Gynecologic and Obstetrics, Ancona University, Italy, were used as study population; 10 women had serous cystoadenoma, 16 women had serous borderline tumor, and 28 women had invasive cystoadenocarcinoma. The expression of primary tumor proliferation related to Ki67 antigen was immunohistochemically evaluated by monoclonal MIB 1 antibody (Immunotech, Marseille Cedex, France) on microwave oven-processed formalin-fixed paraffin-embedded tissue. Compared to cystoadenomas and borderline tumors, the tissular Ki67 antigen immunostaining was significantly higher in cystoadenocarcinomas, with the highest values in architectural grade 2 and 3 neoplasms (P < 0.001). Within the cystoadenocarcinomas, a relationship was observed between the measured proliferation index and disease FIGO stage, but it was not significant (P = 0.92). However, patients who recurred and/or had disease progression presented a primitive neoplasm with significantly higher expression of Ki67 antigen than that of patients with disease-free survival (P = 0.01). A significant relationship was observed between the Ki67 index and disease-free survival, independent of histologic grade and stage, evaluated by Cox hazards analysis (P = 0.02).(ABSTRACT TRUNCATED AT 250 WORDS)

Higher levels of interleukin-6 in cystic fluids from patients with malignant versus benign ovarian tumors correlate with decreased hemoglobin levels and increased platelet counts.

BACKGROUND: Recently, high pretreatment platelet counts and low pretreatment hemoglobin levels were found to be negative prognostic factors in patients with ovarian cancer. Interleukin-6 (IL-6) is a multifunctional cytokine with a diversity of functions leading to the induction of C-reactive protein (CRP), increased platelet counts, and low hemoglobin levels. Different epithelial ovarian cancer cell lines are found to produce varying amounts of IL-6. In this study, a possible relationship between IL-6 levels in cystic fluids of benign and malignant ovarian tumors and pretreatment serum CRP, platelet counts, and hemoglobin levels was evaluated. METHODS: A bioassay and enzyme-linked immunosorbent assay (ELISA) were performed to determine the IL-6 levels in cystic fluids and serum from 42 patients with benign and malignant ovarian tumors. RESULTS: The median IL-6 level was higher in cystic fluids of malignant tumors (n = 21) when compared with cystic fluids of benign tumors (n = 21) (P < 0.01 for bioassay and ELISA). Serum IL-6 levels in patients with malignant tumors were not significantly higher compared with IL-6 levels in patients with benign tumors, whereas CRP levels were higher in patients with malignant tumors (P < 0.01). Cystic fluid IL-6 levels were related to serum CRP levels (r = 0.60, P < 0.01 [bioassay]; r = 0.41, P < 0.01 [ELISA]), and were related inversely to hemoglobin levels (r = -0.57, P < 0.01 [bioassay]; r = 0.54, P < 0.01 [ELISA]). CONCLUSIONS: IL-6 levels are higher in cystic fluids of malignant ovarian tumors compared with benign tumors. The relationship of cystic fluid IL-6 levels with CRP, platelet counts, and hemoglobin levels suggests a possible causative role of tumor-derived IL-6 in the appearance of general side effects of ovarian cancer, which recently have been recognized as prognostic factors.

Serum CA 125 concentrations in patients with benign ovarian tumours.

Serum CA 125 concentrations have been measured in 115 patients with histologically confirmed nonmalignant pelvic disease, that is, serous cystadenoma (n = 56), mucinous cystadenoma (n = 14), fibroma (n = 33), thecoma (n = 8), and Brenner tumour (n = 4). Increased CA 125 concentrations (> 35 KU/L) were found in 14 patients, with a range of 46-891 KU/L, a mean of 205 KU/L, and a median of 97 KU/L. The highest values were found in patients with ascites. Serial measurements in one patient showed a fall in the 2 days immediately after surgery, over the next 3 days showing a two- to three-fold increase, followed by a slow return to normal over the next 7 weeks. Elevated CA 125 levels may not indicate ovarian malignancy and do not differentiate between benign and malignant pelvic masses.

Correlation of serum, urinary and salivary CA 125 levels in patients with adnexal masses.

A prospective study was made of 105 consecutive patients admitted to one department of obstetrics and gynaecology for surgery for adnexal masses. The objective was to investigate if CA 125 level is measurable in the urine or saliva and to correlate these measurements with serum CA 125 level in patients presenting with adnexal masses. The final diagnosis and grouping of patients for analysis were based on histopathological examination of the adnexal masses. Serum, urine and salivary samples were collected simultaneously from all patients on the morning before surgery. CA 125 levels in each sample were determined in duplicate using Abbott CA 125-E1A monoclonal test kits (Abbott Laboratories, USA). The mean inter-assay variability was 10%. CA 125 was detectable in the serum, urine and saliva from all the patients and the concentration was highest in the saliva and lowest in urine. There were no discernible differences in the distributions of salivary CA 125 concentrations between patients with ovarian malignancies and those with benign ovarian cysts. In contrast, both serum and urinary CA 125 levels were significantly higher in the ovarian cancer group. There was no correlation in CA 125 concentrations between serum and urine, or between serum and saliva. For detection of ovarian malignancies, the sensitivity, specificity, and positive and negative predictive values for serum CA 125 measurement (> or = 35 U/mL) were 88.9%, 79.2%, 27.6% and 98.7 respectively. The corresponding figures for urinary CA 125 measurement (> or = 10 U/mL) were 88.9%, 66.7%, 19.5% and 98.4% respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Immunohistochemical study of mucin carbohydrates and core proteins in human ovarian tumors.

Many of the cancer-associated antigens recently have been identified as mucin antigens. However, there are no detailed studies describing the expression of carbohydrates and core proteins of mucin antigens in ovarian tumors. In this study we examined the expression of carbohydrate antigens, which are associated with the earliest steps in mucin glycosylation (Tn and sialosyl-Tn), and the expression of the mucin core protein antigens associated with the MUC1 gene product (mammary-type apomucin) and the MUC2 gene product (intestinal-type apomucin) in 123 ovarian epithelial (mucinous and serous) tumors. In normal ovarian tissues neither Tn, sialosyl-Tn, nor intestinal-MRP antigens (MUC2 gene product) were expressed, except for positive sialosyl-Tn staining of stromal capillaries, while the MUC1 gene product, DF3 antigen, was expressed in the cell apex of the germinal coelomic epithelium when it had plump, slightly elongated, or pseudostratified nuclei. In the benign adenomas Tn and sialosyl-Tn antigens were detected in a small number of mucinous adenomas and rarely in serous adenomas. In contrast, expression of both Tn and sialosyl-Tn antigens was observed in all the adenocarcinomas and in a considerable number of borderline malignancies. DF3 antigen was expressed in many benign serous tumors but not so frequently in benign mucinous tumors; however, it was frequently expressed in the adenocarcinomas and borderline malignancies of both mucinous and serous types. Intestinal-MRP antigen expression increased with the transition of the mucinous tumors from a benign to malignant state, although it was never detected in the serous tumors. Coexpression of DF3 and intestinal-MRP antigens was seen in borderline malignancies and carcinomas of the mucinous tumors. In conclusion, simultaneous expression of Tn and sialosyl-Tn antigens is a highly effective tumor marker in both mucinous and serous tumors of the ovary. Coexpression of DF3 and intestinal-MRP antigens may indicate the malignant potential of ovarian mucinous tumors.

Are CASA and CA125 concentrations in peripheral blood sourced from peritoneal fluid in women with pelvic masses?

BACKGROUND: The Cancer associated serum antigen (CASA) and CA125 assays used in the management of ovarian cancer measure distinct high molecular weight glycoproteins. The mechanisms of secretion of these molecules into the peripheral circulation are not clearly understood. METHODS: Concentrations of CASA and CA125 were assessed in peripheral blood, blood from veins draining ovarian and omental tumors, and peritoneal fluid in 20 women with pelvic masses. RESULTS: There was a near perfect correlation between peripheral vein and ovarian vein concentrations for both markers; concentrations in omental veins were higher than in peripheral veins in only a small proportion of cases, whereas the concentration in peritoneal fluid was universally much higher than in blood. CONCLUSIONS: These studies suggest a similar route of entry into the peripheral circulation and a similar half-life for these glycoproteins. These data provide no support for the hypothesis that a significant contribution to CA125 concentrations comes from peritoneal release by mesothelial cells rather than tumor cells, because the relative CA125 concentrations between compartments were similar to those of CASA within patients. The lack of a gradient between CASA levels in peripheral blood and tumor draining veins suggests that the CASA half-life is much longer than that predicted in animal studies. CA125 and CASA in peripheral blood are probably derived from markers secreted into peritoneal fluid and lymph, rather than directly into the bloodstream.

Relation between ovarian carcinoma-associated antigens in tumor tissue and detached cyst fluid cells of patients with ovarian neoplasms.

AIMS: The expression and potential diagnostic value of ovarian carcinoma-associated antigens were estimated in different types of epithelial ovarian neoplasms. The comparison of antigenic expression was performed on solid tumor tissues and loose cyst fluid cells in individual cases of malignant and benign ovarian neoplasms. METHODS: All studies were performed using monoclonal antibodies (mAbs) against ovarian carcinoma-associated antigens (OC125, OV-TL3, OV632, 10B, 8C) by 3-step peroxidase-antiperoxidase test. RESULTS: All ovarian carcinoma-associated antigens were detected in most serous and endometrioid carcinomas. In mucinous carcinomas as well as in benign ovarian neoplasms these antigens were present only in some cases. Significant inter- and intratumoral immunological heterogeneity was evident; however, the antigens detectable in tissue sections were also found in detached cyst fluid cells. CONCLUSIONS: Our results show that mAb show OV-TL3 is the best marker for endometrioid carcinomas and confirmed that mAbs OV632, OC125 and OV-TL3 could be good complementary markers for differentiating malignant and benign lesions in the ovary. The percentage content of all ovarian carcinoma-associated antigens in solid tumors and respective cyst fluid cells was comparable.

Characterization of OM-B monoclonal antibody-defined antigen associated with mucinous type human ovarian tumor.

Partial characterization of the OM-B antigen associated with mucinous-type ovarian tumors was conducted. This antigen was defined by OM-B monoclonal antibody, which was raised against a mucinous-type ovarian tumor, and was present in all the mucinous-type tumors tested, but only a fraction of serous-type tumors. The OM-B crude antigen preparation fractionated from cystic fluids had a density of 1.40-1.43 g/ml, with a high neutral sugar content. Molecular mass (M(r)) estimated by gel filtration was more than 2,000,000. Trypsinization of the antigen preparation under appropriate conditions resulted in two major bands and one minor band with molecular sizes of less than M(r) 250,000, as detected by immunoblotting. Immunoaffinity chromatography was then conducted and the amino acid composition of the purified product was determined; the high contents of serine, threonine and proline are characteristic of a mucin. Binding inhibition enzyme-linked immunosorbent assay was developed to measure OM-B antigen activity in cystic fluids and sera from patients with mucinous-type tumors. The antigen was easily detected in most cystic fluids, but not in sera, suggesting that improvement in the sensitivity of this assay is necessary before its utilization for serum diagnosis will be feasible.

Natural killer cell activity and progression-free survival in ovarian cancer.

A longitudinal evaluation of the natural killer (NK) cell activity of peripheral blood lymphocytes was performed in 17 patients with advanced ovarian cancer (treated surgically and with subsequent multiagent chemotherapy) for a median follow-up period of 32 months. At the time of primary treatment, the mean value of NK cell activity was significantly lower in patients who then had disease progression (p < 0.05) than in patients with progression-free survival. During the follow-up period, no significant modifications of the NK cell activity were observed; only at the time of clinical disease progression did the NK cell activity show a significant reduction. We conclude that the NK cell activity is a potentially important prognostic factor.

Tumor-infiltrating lymphocytes from ovarian tumors of low malignant potential.

Tumor-infiltrating lymphocytes (TIL) from human ovarian tumors of low malignant potential (LMP) were studied in situ by immunohistology and characterized functionally after isolation from tumors. In comparison to ovarian carcinomas, borderline (LMP) ovarian tumors contained significantly fewer infiltrating leukocytes. Phenotypically, TIL from LMP tumors contained significantly fewer activated (HLA-DR+ or CD25+) lymphocytes than did fresh TIL from ovarian carcinomas. Also, percentages of CD3-CD56+ natural killer cells were higher in LMP than in ovarian carcinomas. TIL obtained from LMP tumors proliferated well in vitro in the presence of interleukin 2 (IL2) and tumor necrosis factor alpha (TNF alpha) but did not show a selective enrichment in CD3+CD8+T lymphocytes generally observed with TIL from ovarian carcinomas. Antitumor cytotoxicity against a panel of tumor cell targets of cultured TIL from LMP tumor did not parallel that of effectors generated from autologous peripheral blood lymphocytes. However, no significant enrichment in reactivity against autologous tumor cells was observed in long-term cultures of TIL from LMP tumors. Thus, considerable differences were observed between phenotypic and functional characteristics of lymphoid cells infiltrating LMP tumors and invasive ovarian carcinomas.

Carcinoembryonic antigen isotypes in tissue sections and loose cyst fluid cells of ovarian neoplasms.

The aim of this study was to establish whether different subsets of ovarian neoplasms express a restricted isotype of carcinoembryonic antigen (CEA) which can be detected in solid tumors and detached cells. Sixty-one cases of mucinous, serous, endometrioid, and Krukenberg tumors were studied by immunohistochemistry using two monoclonal antibodies (MAbs), commercial anti-CEA and D14 with a higher specificity for colorectal adenocarcinomas. The results with both antibodies showed a considerable degree of heterogeneity between cases of nonserous tumors, with a more restrictive pattern observed with the D14 MAb. The proportion of immunostained cells was comparable in tumors and fluids.

Elevated serum CA 19-9 levels in hepatobiliary cystadenoma with mesenchymal stroma. Two case reports with immunohistochemical confirmation.

Hepatobiliary cystadenomas with mesenchymal stroma (CMS) are rare tumors. Two cases are reported that illustrate many features of CMS, including polypoid involvement of the common hepatic duct in one case. Both tumors were associated with elevated serum levels of the tumor-associated antigen CA 19-9 but normal levels of carcinoembryonic antigen and alpha-fetoprotein. Immunohistochemical analysis confirmed the presence of CA 19-9 in the epithelial component of the tumor. The implications of these findings are discussed, both in relation to the histogenesis of CMS and its preoperative diagnosis. A minimally elevated level of CA 19-9 was found in only one of five patients with hydatid disease of the liver, an important differential diagnosis in clinical management.

Some considerations on the biology of pancreatic serous cystadenoma.

Five cases of pancreatic serous cystadenoma were examined pathologically, and their nuclear DNA ploidy patterns were determined. Four were unifocal tumors, and one was a multifocal tumor. The four unifocal tumors were typical serous cystadenomas. However, the multifocal tumor exhibited an increased N/C ratio, irregular nuclear margins, various-sized nuclei, coarse nuclear chromatin, and neural invasion. All tumor cells were stained with antiCA19-9 but none with antiCEA. In the antiCA19-9 staining, the four unifocal tumors and the tumors of the pancreatic tail in the multifocal case were positive only on the apical membrane, whereas the tumor cells of the pancreatic head in the multifocal case were positive within the whole cytosol. The unifocal tumors were diploid with a DNA Index (DI) of 1.0 and proliferation indices (PI) from 4.9 to 20.9% with a mean of 14.4%. In the multifocal case, the tumor in the pancreatic head was aneuploid (DI = 1.9) and had a PI of 27.8%. The multifocal sites in the pancreatic body were aneuploid (DI = 1.9) with a PI of 22.4%. We suggest that the biological property of serous cystadenoma should be revisited.

Immunohistochemical study of carcinoembryonic antigen in mucinous cystic neoplasm of the pancreas.

Pathologic and immunohistochemical studies of 13 patients with mucinous cystic neoplasm of the pancreas were performed in order to determine whether these lesions represent overt or latent malignancy. In 9 patients with mucinous cystadenoma, the foci of mucus-rich columnar epithelia formed the usual pattern of stratifications and papillary projections with a benign appearance and did not stain with mouse monoclonal antibody to CEA (CM-010, Mochida, Japan). In contrast, the foci of mucus-poor columnar epithelia which were in a focal distribution showed moderate to severe atypical hyperplasia and reacted strongly with anti-CEA in 6 of the 9 patients. In addition, areas of both noninvasive and invasive mucinous cystadenocarcinomas in 4 patients, which were composed primarily of mucus-poor columnar epithelia, showed diffuse cytoplasmic and/or apical reactivity with anti-CEA. Based on these results, we conclude that severe atypical epithelial hyperplasia composed primarily of mucus-poor columnar epithelia represents a precursor to mucinous cystadenocarcinoma, and, therefore, mucus-poor atypical epithelia may undergo malignant transformation.