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.

Evolution of the expression of fetal acinar antigens during carcinogenesis of the pancreas in hamsters: individual follow-up by open biopsy.

Individual lesions in the pancreas and the presence of fetal acinar antigens along with carcinogenesis induced by N-nitrosobis(2-oxopropyl)amine (CAS: 60599-38-4) were studied by open biopsy in 16 Syrian golden hamsters 13, 22, and 40 weeks after initiation of treatment. At 13 weeks, cystadenoma and regular ductal hyperplasia were noted in 3 animals and 1 animal, respectively. Staining for fetal acinar antigens in the pancreas was found in 69% of the hamsters. At 22 weeks, cystadenoma and hyperplastic ducts were common (60 and 53%), and 3 hamsters developed pancreatic adenocarcinoma. Fetal acinar antigens persisted in the acini and extended to irregular hyperplastic ducts and tumor cells. At 40 weeks, ductal proliferation was the main lesion in all pancreatic tissue, and 9 animals had adenocarcinoma. Acinar antigens were found in the remaining acini, in irregular hyperplastic ducts, and in tumor cells. Thus, once reexpressed, fetal acinar antigens persist in pancreatic lesions and pancreatic carcinomas in the hamster.

Carcinoembryonic antigen in human ovarian neoplasms.

The presence and location of carcinoembryonic antigen (CEA) was examined in tissue sections of 14 mucinous and 13 serous cystadenomas and cystadenocarcinomas of the ovary. CEA was demonstrated in the mucinous tumors but was not present in the serous tumors. In the mucinous tumors, CEA was located in the glycocalyx and apical portions of the absorptive-type epithelium, with only trace quantities in goblet cells, a pattern identical to that seen in colonic neoplasms. Endocervical-type epithelium in the mucinous tumors contained little or no CEA.

Expression of cell-mediated immunity and blocking factor using a new line of ovarian cancer cells in vitro.

A cell-mediated cytotoxicity test, quantitated by postlabeling with tritiated thymidine, was used to asses immune reactivity of cancer patients to the HeW cell line derived from serous cystadenocarcinoma of the ovary. Lymphocytes from 71.4% of serous and mucinous cystadenocarcinoma patients demonstrated a cytotoxic response towards the HeW cells, whereas no reactivity was observed towards target cells derived from nonovarian cancer. These observations indicated that the HeW cells express tumor-associated antigen. In some patients bearing similar tumors, cytotoxicity was blocked by ascitic fluid from other patients with cystadenocarcinoma. In addition, antigen obtained from the spent culture fluid of HeW cells exhibited blocking activity in a typical dose-response fashion, suggesting that blocking factor may be free tumor-associated antigen or an antigen-specific suppressor molecule. Thus, blocking of the lymphocytotoxic response of cystadenocarcinoma patients towards HeW cells may be utilized to monitor the isolation of ovarian carcinoma-associated antigen.

Characterization of the carcinoembryonic antigen activity associated with cyst fluids of mucinous ovarian cystadenocarcinoma.

Cyst fluids from mucinous cystadenocarcinoma of the ovary show significant concentrations of carcinoembryonic antigen (CEA) activity. This CEA activity was compared to several colon CEA standards with respect to size, concanavalin A binding, and immunological activity. CEA activity in unfractionated ovarian cyst fluid was indistinguishable from CEA standards by gel filtration chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The cyst fluid CEA activity showed identity with various CEA standards by double immunodiffusion in agar gels and by radioimmune competition assay. Attempts to purify the cyst fluid CEA by perchloric acid extraction prior to gel filtration chromatography were unsuccessful. The yield of soluble CEA activity following perchloric acid extraction averaged 10%, and in one cyst fluid, qualitative changes were noted in the soluble antigenic activity. Ovarian cyst fluid CEA activity was bound by concanavalin A-Sepharose 4B and specifically eluted with competing monosaccharide. Lectin affinity chromatography and gel filtration chromatography over Sepharose 4B and Sephadex G-200 can be used to purify cyst fluid CEA from whole fluid or lyophilized and reconstituted samples.

A study of cyst fluid and plasma carcinoembryonic antigen in patients with cystic ovarian neoplasms.

Cyst fluid and plasma carcinoembryonic antigen (CEA) levels were measured in 11 patients with ovarian cyst-adenocarcinoma and in 16 patients with benign ovarian neoplasms. In patients with ovarian cancer, plasma CEA levels were not elevated above 2.5 ng/ml unless cyst fluid CEA levels were 4 to 16 mu-g/ml. In this series, cystic and plasma CEA levels were elevated most consistently in patients with mucinous ovarian tumors. Furthermore, on the basis of molecular size and immunoreactivity by immunodiffusion, ovarian cancer cyst fluid CEA and colonic cancer CEA had similar immunochemical properties. Consistent with the findings in other neoplasms, follow-up studies showed that plasma CEA levels returned to the normal range between 2 and 12 weeks after surgical excision of the ovarian tumor. It is concluded that plasma CEA is of value in the management of patients with ovarian mucinous cystadenocarcinoma.

Monoclonal antibodies that discriminate between human ovarian carcinomas and benign ovarian tumours.

Three hybridoma cell lines producing monoclonal antibodies (MAbs) against ovarian carcinomas were obtained after immunizing mice with an undifferentiated human ovarian cystadenocarcinoma extract. The hybridoma cell supernatants were initially screened for a positive immunohistochemical reaction with ovarian carcinomas concomitant with a negative reactivity with a benign ovarian cystadenoma. The antibodies OV-TL 15 (IgM), OV-TL 30 (IgG1) and OV-TL 31 (IgM) reacted positively with 84, 84 and 74% of the ovarian carcinoma samples (n = 76) respectively. Reactivity with ovarian cysts and cystadenomas (n = 21), with non-ovarian carcinomas (n = 77) and with normal human tissue samples (n = 63) was absent or limited to weak reactivity on incidental samples. All three antibodies were shown by immunoelectron microscopy to react with surface antigens (OA 15, OA 30 and OA 31) of ovarian carcinoma cells. Cross reactivity between OV-TL 15, OV-TL 30, OV-TL 31 and OC 125 monoclonal antibodies was excluded by competitive binding assays on NIH:OVCAR-3 cells. Antigen levels (OA 15 and OA 31) in tumour extracts and cyst fluids were quantified by immunoradiometric assays and compared to the CA 125 antigen levels. High levels of CA 125, OA 15 and OA 31 were found in cyst fluids from ovarian cancers. In benign ovarian cyst fluids, however, the CA 125 content was also high while OA 15 antigen was hardly detectable. The OA 31 antigen was present at relatively low levels. The IRMA data confirmed the immunohistochemical data showing that, in contrast to OC 125, the newly developed MAbs OV-TL 15, OV-TL 30 and OV-TL 31 discriminate between benign ovarian cystadenomas and malignant ovarian cancers.

Detection of carcinoembryonic antigen in tissue sections by immunoperoxidase.

A triple-bridge, indirect, immunoperoxidase method for detecting and localizing carcinoembryonic antigen (CEA) in tissue sections is described. By this technique, a cell-surface localization of CEA in colonic carcinoma and ovarian mucinous cystadenocarcinoma cells could be visualized. In the case of the colonic cancer, both the tumor from the descending colon and a metastasis to the skin gave positive peroxidase reactions for CEA. This immunocytochemical method for demonstrating the presence of CEA functioned in both frozen, ethanol-fixed and formalin-fixed, paraffin-embedded tissues, thus making it applicable for use with tissue sections conventionally prepared for light microscopy.

Intratesticular serous papillary cystadenoma of low malignant potential: an ultrastructural and immunohistochemical study suggesting müllerian differentiation.

A 51-year-old man was found to have two separate intratesticular serous cystadenomas of low malignant potential. Electron microscopy and OC 125 antibody reactivity characterize the neoplasms as serous and suggest müllerian histogenesis.

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.

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.

Expression of CEA, CA125, CA19-9 and human milk fat globule membrane antigen in ovarian tumours.

The expression of five different antigens in ovarian tumours was studied by means of an immunohistochemical test with anti-CEA, HMFG1 and HMFG2, NS19-9 and OC125 antibodies. Considerable variation was noted not only between different histological types and between tumours of one type but also between areas in a single tumour. HMFG1 and HMFG2 were the most reactive of all the antibodies; NS19-9 and OC125 were expressed by different populations of cells. It is concluded that specific combinations of antibodies are more effective both for the monitoring of ovarian cancer as well as for immunodiagnosis and treatment, than any single one used.

Morphological and immunological heterogeneity of human ovarian neoplasms.

The cells of cystic fluids from patients with malignant and benign serous ovarian neoplasms were separated by density gradient centrifugation. Density distribution and morphological features of cell fractions were analyzed. The immunophenotypic characterization of each cell fraction using poly- and monoclonal antibodies was performed. The application of density gradient centrifugation allowed to yield and catalogue several cell subpopulations according to their stage of pathological differentiation starting from typical, morphologically malignant until normal epithelial cells. Significant morphological and immunological heterogeneity among and within individual tumors was found. Our studies indicated the value of isolated cell subsets for estimation of interrelationships among various neoplastic markers and for comparison the reactivity of different antibodies and evaluation of their immunodiagnostic potency.

Density distribution and antigenic characterization of cell subpopulations isolated from cystic fluids of ovarian serous neoplasms.

The immunological reactivity of serous ovarian tumor cells was evaluated, taking into account their density and morphological features. Discontinuous density gradient centrifugation was applied to fractionate cell subpopulations from cystic fluids of patients with ovarian serous carcinomas, cystadenomas and benign serous cysts. For phenotypic characterization of tumor cell subpopulations the immune sera against perchloric acid extracts of ovarian serous (anti-PCA-CaOs) and mucinous (anti-PCA-CaOm) carcinomas were used. The expression of carcinoembryonic antigen (CEA) and nonspecific cross-reacting antigen (NCA) on tumor cell fractions was also checked. Some relationship between immunological reactivity, cell morphology and cell density was found; however, individual patient-to-patient variations in cellular composition and antigenic expression were also observed. The presence of ovarian serous carcinoma-associated antigen (CaOs-Ag) was related mainly to frankly malignant cells. Neither CEA nor NCA were constant and characteristic markers for serous ovarian neoplasms. These neoplasms were unreactive with anti-PCA-CaOm serum. Our results indicate the possibility of correlation between morphological features, density distribution and immunological phenotypes of ovarian tumor cell populations.

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.

Intestine-associated antigens in ovarian tumours: an immunohistological study.

The presence of 3 intestine-associated antigens, small intestine mucin antigen (SIMA), large intestine mucin antigen (LIMA) and carcinoembryonic antigen (CEA) was studied in the female genital tract and ovarian tumours by immunofluorescence. These antigens could not be detected in normal ovary, benign cysts of ovary, fallopian tube or endometrium, but both LIMA and CEA were present in endocervical glandular tissue. The antigenic cross-reactivity of endocervical and large bowel mucin may indicate a close embryological relationship between these organs during the cloacogenic stage. The 3 antigens could be demonstrated in mucinous tumours of the ovary but were absent in serous or mesonephroid tumours. In one of the 2 endometroid tumours CEA was the only detectable antigen. These observations confirm the presence of intestinal type of epithelium in cystic mucinous tumours of the ovary and explain the cross-reactivity of mucin of benign tumours of the ovary and mucin from colonic cancer, normal colonic mucosa and gastric mucosa as reported by earlier workers. In the process of malignant transformation the columnar epithelium of ovarian cystadenoma seems to behave in the same way as superficial gastric and gall bladder epithelium by forming inappropriate intestine-associated mucin substances. Our technique may provide a specific means for studies on the histogenesis of female genital tract tumours, particularly ovarian tumours. It can also be used in differentiating between benign and malignant variants of these tumours.

An ovarian tumour specific mucin antigen--immunohistological and biochemical studies.

A new mucin antigen was detected in a mucinous cystadenoma of human ovary. An antiserum produced in rabbits against the crude cyst fluid, following appropriate absorptions, showed immunoreactivity exclusively with tumour epithelium, particularly endocervical type epithelium. The antigen responsible for the immunoreactivity was isolated in the fraction with density 1.45g/ml following density gradient ultracentrifugation in cesium chloride, which indicated that it was indeed a glycoprotein. The native glycoprotein was very large since it was excluded from Sepharose CL-2B. The complex could be dissociated following reduction with dithiothreitol. The subunits were included in Sepharose CL-2B and the position of elution would indicate a molecular weight in the order of 1-5 x 10(6). This suggests the native antigen was a complex made up of subunits held together by disulphide bonds. Amino acid analysis of the new antigen showed a resemblance with intestinal mucins, as two-thirds of the protein consists of threonine, serine, proline, alanine and glycine. This similarity in peptide core may explain the potential of the epithelium in ovarian mucinous cystadenoma to produce inappropriate intestinal mucins during the process of malignant transformation.

Mucinous cystic neoplasm of the pancreas with high carcinoembryonic antigen.

A large mucinous cystic neoplasm of the pancreas was found that showed a remarkably high carcinoembryonic antigen (CEA) level in cyst content. Staining for CEA was also detected in the lining epithelium. These findings indicate that columnar epithelial cells secrete CEA that accumulates in the cyst. We suggest that a CEA level determination of the cyst fluid, along with CEA immunoperoxidase staining on cell preparations, may prove useful in providing an accurate diagnosis of pancreatic cystic tumors.

Tumor and serum CEA content in gynecologic neoplasms.

The relationship of serum carcinoembryonic antigen (CEA) levels to tumor tissue CEA content, and tumor weight in patients with different genital tract neoplasms was studied. Two-step extraction of tumor tissues was performed using perchloric acid and 3 M KCl. Sera and tissue extracts were assayed for CEA content by double-antibody radioimmunoassay. No clear correlation was found between serum CEA and tumor CEA contents, tumor weight, and histological structure. Our results showed, however, that high CEA concentrations in tumor tissue (10 micrograms/g or higher) were accompanied by elevated values in plasma.

Detection of tumor-specific antigens in human mucinous cystadenocarcinoma of the ovary by immunodiffusion.

Rabbit antiserum to a tissue extract of human mucinous cystadenocarcinoma of the ovary reacted with tissue extracts of normal ovary and various ovarian malignancies, and ascitic or cystic fluids of ovarian origin by Ouchterlony double gel diffusion and precipitin inhibition techniques. The tumor-associated antigen(s) of mucinous cystadenocarcinoma, which were demonstrated by Ouchterlony double diffusion, were not present in tissue extract of pooled normal ovaries and cystic fluid of benigh tubo-ovarian cyst. An organ-associated tumor antigen as well as the type-specific tumor antigen may exist in mucinous cystadenocarcinoma of the ovary. The mucinous cystadenocarcinoma was not very immunologically different but was distinguishable from serous cystadenocarcinoma and other types of ovarian cancer by double gel diffusion. Precipitin-inhibition reactions demonstated that the adsorbed antiserum to human ovarian mucinous cystadenocarcinoma mixed with tissue extracts of dysgerminoma and serous cystadenocarcinoma, and ascitic fluid of papillary embryonal adenocarcinoma of the ovary could not eliminate the specific precipin line developed with tissue extract of mucinous cystadenocarcinoma.