cefoxitin and Adenocarcinoma

Mucin-type O-glycans are involved in cancer initiation and progression, although details of their biological and clinicopathological roles remain unclear. The aim of this study was to investigate the clinicopathological significance of β1,3-N-acetylglucosaminyltransferase 6 (β3Gn-T6), an essential enzyme for the synthesis of core 3 O-glycan and several other O-glycans in pancreatic ductal adenocarcinoma (PDAC). We performed immunohistochemical and lectin-histochemical analyses to detect the expression of β3Gn-T6 and several O-glycans in 156 cases of PDAC with pancreatic intraepithelial neoplasias (PanINs) and corresponding normal tissue samples. The T antigen, Tn antigen, sialyl Lewis X (sLeX) antigen, and sLeX on core 2 O-glycan were more highly expressed in PDAC cells than in normal pancreatic duct epithelial cells (NPDEs). Conversely, the expression of 6-sulfo N-acetyllactosamine on extended core 1 O-glycan was found in NPDEs and was low in PDAC cells. These glycan expression levels were not associated with patient outcomes. β3Gn-T6 was expressed in ~20% of PDAC cases and 30-40% of PanINs but not in NPDEs. Higher expression of β3Gn-T6 was found in PDAC cells in more differentiated adenocarcinoma cases showing significantly longer disease-free survival in both univariate and multivariate analyses. In addition, the expression of β3Gn-T6 in PDAC cells and PanINs significantly correlated with the expression of MUC5AC in these cells, suggesting that β3Gn-T6 expression is related to cellular differentiation status of the gastric foveolar phenotype. Thus, it is likely that β3Gn-T6 expression in PDAC cells is a favorable prognostic factor in PDAC patients, and that the expression of β3Gn-T6 correlates with the gastric foveolar phenotype in pancreatic carcinogenesis.

Topics: Adenocarcinoma; Adult; Aged; Aged, 80 and over; Antigens, Tumor-Associated, Carbohydrate; Antigens, Viral, Tumor; Carcinoma, Pancreatic Ductal; Female; Gene Expression Regulation, Neoplastic; Humans; Male; Middle Aged; N-Acetylglucosaminyltransferases; Polysaccharides; Sialyl Lewis X Antigen

Human pancreatic ductal adenocarcinoma (PDAC) is one of the most aggressive and lethal malignancies in the world and despite great efforts in research types of treatment remain limited. A frequently detected alteration in PDACs is a truncated O-linked N-acetylgalactosamine (GalNAc) glycosylation with expression of the Tn antigen. Changes in O-glycosylation affect posttranslationally modified O-GalNAc proteins resulting in profound cellular alterations. Tn antigen is a tumor associated glycan detected in 75-90 % of PDACs and up to 67 % in its precursor lesions. Since the role of Tn antigen expression in PDAC is insufficiently understood we analyzed the impact of COSMC mediated Tn antigen expression in two human PDAC cell lines on cellular oncogenic properties.. Forced expression of Tn antigen on O-glycosylated proteins in pancreatic cancer cells was induced by lentiviral-mediated knockdown of the COSMC chaperone, which prevented O-glycan elongation beyond the initial GalNAcα1- residue on O-linked glycoproteins. Altered O-GalNAc glycosylation was analyzed in human pancreatic cancer cell lines Panc-1 and L3.6pl using Western and Far-Western blot as well as immunocytochemical techniques. To assess the biological implications of COSMC function on oncogenic properties, cell viability assays, scratch assays combined with live cell imaging, migration and apoptosis assays were performed. Lectin based glycoprotein enrichment with subsequent mass spectrometric analysis identified new cancer O-GalNAc modified proteins. Expression of Tn antigen bearing Nucleolin in patient derived PDAC tumor specimens was evaluated and correlated with clinicopathological data.. Tn antigen expression was induced on various O-GalNAc glycoproteins in COSMC deficient cell lines compared to the control. Proliferation was reduced (p < 0.001) in COSMC knockdown cells, whereas migration was increased (p < 0.001) and apoptosis was decreased (p = 0.03), highlighting the importance of Tn antigen expression on metastatic and anti-apoptotic behavior of PDAC derived cells. Nucleolin was identified as O-GalNAc modified protein in COSMC deficient PDAC cell lines. Interestingly, immunohistochemical staining and co-localization studies of patient derived PDACs revealed poor survival for patients with strong co-localization of Tn antigen and Nucleolin (p = 0.037).. This study substantiates the influence of altered O-glycan (Tn/STn) expression on oncogenic properties in pancreatic cancer and identifies O-GalNAc modified Nucleolin as novel prognostic marker.

Topics: Adenocarcinoma; Antigens, Tumor-Associated, Carbohydrate; Carcinogenesis; Carcinoma, Pancreatic Ductal; Cell Line, Tumor; Cell Movement; Cell Proliferation; Cell Survival; Gene Expression Regulation, Neoplastic; Gene Knockdown Techniques; Glycosylation; Humans; Mass Spectrometry; Molecular Chaperones; N-Acetylgalactosaminyltransferases; Neoplasm Proteins; Nucleolin; Pancreatic Neoplasms; Phosphoproteins; Polysaccharides; RNA-Binding Proteins; RNA, Messenger

The Tn antigen, which arises from mutation in the Cosmc gene is one of the most common tumor associated carbohydrate antigens. Cosmc resides in X24 encoded by a single gene and functions as a specific molecular chaperone for T-synthase. While the Tn antigen cannot be detected in normal cells, Cosmc mutations inactivate T-synthase and consequently result in Tn antigen expression within certain cancers. In addition to this Cosmc mutation-induced expression, the Tn antigen is also expressed in such cell lines as Jurkat T, LSC and LS174T. Whether the Cosmc mutation is present in the colon cancer cell line HT-29 is still unclear. Here, we isolate HT-29-Tn+ cells from HT-29 cells derived from a female colon cancer patient. These HT-29-Tn+ cells show a loss of the Cosmc gene coding sequence (CDS) leading to an absence of T-synthase activity and Tn antigen expression. Additionally, almost no methylation of Cosmc CpG islands was detected in HT-29-Tn+ as well as in HT-29-Tn- and Tn- tumor cells from male patients. In contrast, the methylation frequency of CpG island of Cosmc in normal female cells was ~50%. Only one active allele of Cosmc existed in HT-29-Tn+ and HT-29-Tn- cells as based upon detection of SNP sites. These results indicate that Tn antigens expression and T-synthase inactivity in HT-29-Tn+ cells can be related to the absence of CDS in Cosmc active alleles, while an inactive allele deletion of Cosmc in HT-29 cells has no influence on Cosmc function.

Topics: Adenocarcinoma; Antigens, Tumor-Associated, Carbohydrate; Cell Separation; Colorectal Neoplasms; Flow Cytometry; Fluorescent Antibody Technique; Gene Expression Regulation, Neoplastic; HT29 Cells; Humans; Molecular Chaperones; Mutation; Reverse Transcriptase Polymerase Chain Reaction

Overexpression of mucins is known to decrease cell-to-cell adhesion and thus to facilitate the invasion of cancer cells through the extracellular matrix. Mucin 6 (MUC6) is overexpressed and aberrantly O-glycosylated in human breast cancer, serving as a carrier for one of the most specific cancer-associated antigens, Tn antigen. Despite its relevance in breast cancer, MUC6 expression has not yet been characterized in canine mammary tumours (CMTs). The aims of this study were to assess the expression of MUC6 and Tn antigen in 55 benign and 77 malignant CMTs of different histological types and to investigate possible correlations with pathological features. MUC6 and Tn antigen were found to be significantly overexpressed in malignant compared with benign CMTs. MUC6 was significantly overexpressed in simple and complex carcinomas compared with simple and complex adenomas, respectively. When considering only the epithelial population, significant MUC6 overexpression was observed in carcinosarcomas when compared with benign mixed tumours. In addition, MUC6 was significantly overexpressed in simple compared with complex carcinomas. Finally, double-labelling immunofluorescence performed on seven malignant CMTs showed MUC6 and Tn co-expression. Therefore, MUC6 and Tn antigen overexpression is associated with malignant phenotypes of CMTs.

Topics: Adenocarcinoma; Adenoma; Animals; Antigens, Tumor-Associated, Carbohydrate; Biomarkers, Tumor; Carcinosarcoma; Dog Diseases; Dogs; Female; Immunohistochemistry; Mammary Glands, Animal; Mammary Neoplasms, Animal; Microscopy, Fluorescence; Mucin-6

The tumor-associated Tn antigen has been investigated extensively as a biomarker and therapeutic target. Cancer vaccines containing the Tn antigen as a single tumor antigen or as a component of a polyvalent vaccine have progressed into phase I and II clinical trials. One major focus of Tn-based vaccines is the treatment of prostate cancer patients. Although expression of the antigen on prostate tumors is a critical prerequisite, previous reports investigating Tn expression in prostate tumors have produced conflicting results. Using a combination of immunohistochemistry and carbohydrate microarray profiling, we show that only 4% to 26% of prostate tumors express the Tn antigen. Based on our results, the majority of prostate cancer patients do not express the appropriate antigen. Therefore, efforts to preselect the subset of prostate cancer patients with Tn-positive tumors or apply Tn vaccines to other cancers with higher rates of antigen expression could significantly improve clinical response rates. Because conflicting information on carbohydrate expression is a general problem for the field, the approach described in this article of analyzing antigen expression with multiple antibodies and using carbohydrate microarray profiles to interpret the results will be useful for the development of other carbohydrate-based cancer vaccines and diagnostics.

Topics: Adenocarcinoma; Aged; Animals; Antigens, Tumor-Associated, Carbohydrate; Biomarkers, Tumor; Cancer Vaccines; Carbohydrates; Carcinoma, Transitional Cell; Clinical Trials as Topic; Humans; Hyperplasia; Male; Prostatic Neoplasms; Rabbits

The epithelial mucin MUC1 is a high molecular weight membrane glycoprotein frequently overexpressed and aberrantly glycosylated in adenocarcinoma. Mucins normally contain high amounts of O-linked carbohydrate structures that may influence immune reactions to this antigen. During malignant transformation, certain glyco-epitopes of MUC1, such as Tn-antigen, TF-antigen and their sialylated forms become exposed. The role of these glycan structures in tumor biology is unknown, but their presence is known to correlate with poor prognosis in several adenocarcinomas. We analyzed the potency of MUC1 containing Tn-antigens (MUC1-Tn) to target C-type lectins that function as carbohydrate recognition and uptake molecules on dendritic cells (DC). We identified the macrophage galactose type C-type lectin (MGL), expressed by both DC and macrophages, as the receptor for recognition and binding of MUC1-Tn. To validate the occurrence of MGL-MUC1 interactions in situ, we studied the binding of MGL to MUC1 in primary colon carcinoma tissue. Isolation of MUC1 out of colon carcinoma tissue showed strong binding activity to MGL. Interestingly, MGL binding to MUC1 was highly correlated to binding by the lectin Helix pomatia agglutinin (HPA), which is associated with poor prognosis in colorectal cancer. The detection of MGL positive cells in situ at the tumor site together with the modified glycosylation status of MUC1 to target MGL on DC suggests that MGL positive antigen presenting cells may play a role in tumor progression.

Topics: Acetylgalactosamine; Adenocarcinoma; Animals; Antigens, Neoplasm; Antigens, Tumor-Associated, Carbohydrate; Cell Adhesion Molecules; CHO Cells; Colon; Colonic Neoplasms; Cricetinae; Cricetulus; Dendritic Cells; Endocytosis; Female; Glycosylation; Humans; Immunoglobulin G; Intestinal Mucosa; Lectins, C-Type; Mice; Mice, Inbred BALB C; Monocytes; Monosaccharides; Mucin-1; Mucins; Neoplasm Proteins; Protein Binding; Protein Processing, Post-Translational; Protein Structure, Tertiary; Receptors, Cell Surface; Recombinant Fusion Proteins; Tandem Repeat Sequences

Tumor-associated antigens are appreciated as diagnostic markers, but they have also prompted tremendous efforts to develop tumor-specific immunotherapy. A previously cloned tumor-associated antigen, EBAG9, was initially defined by reactivity with the monoclonal antibody 22-1-1. Functionally, the EBAG9-encoded gene-product was believed to induce apoptosis in activated immune cells. However, using a cell-biological approach we identified EBAG9 as a Golgi-resident modulator of O-linked glycan expression, the latter product was then recognized by the 22-1-1 antibody. Secondly, EBAG9 expression was found physiologically in all murine tissues examined. This raised the question if EBAG9 is tumor-specific and mediates apoptosis itself or through O-linked glycans generated, among them the cognate 22-1-1 antigen Tn.. We have used immunohistochemistry to detect the expression of 22-1-1 and EBAG9 in various tissues. Correlation between expression of both antigens in cell lines was analysed by immunoblot and flow cytometry. Apoptosis was studied by using flow cytometry and Caspase-Glo 3/7 assay kit. Cellular distribution of EBAG9 was analysed by electron and confocal microscopy.. Here, we compared expression of the 22-1-1 and EBAG9-defined antigens in normal and neoplastic tissues in situ. In contrast to 22-1-1 staining, EBAG9 is a ubiquitously expressed antigen in all normal and cancerous tissues. Functional studies on the role of 22-1-1 reactive material did not support any evidence for apoptosis induction. Employing electron and confocal microscopy, a refined subcellular localization of EBAG9 at the Golgi was obtained.. We suggest that the estrogen-inducible EBAG9 gene-product and the 22-1-1 defined antigen are structurally and functionally separate antigens.

Topics: Adenocarcinoma; Antibodies, Monoclonal; Antigens, Neoplasm; Antigens, Tumor-Associated, Carbohydrate; Antineoplastic Agents; Apoptosis; Biomarkers, Tumor; Brefeldin A; Carcinoma; Carcinoma, Squamous Cell; Caspase 3; Caspase 7; Caspases; Cell Line; Cell Line, Tumor; Cell Nucleus; Colorectal Neoplasms; Flow Cytometry; Golgi Apparatus; Humans; Immunoblotting; Immunohistochemistry; Immunotherapy; Lung Neoplasms; Male; Microscopy, Confocal; Microscopy, Electron; Mouth Neoplasms; Nocodazole; Polysaccharides; Prostatic Neoplasms; Protein Synthesis Inhibitors; Reverse Transcriptase Polymerase Chain Reaction; Stomach Neoplasms; Subcellular Fractions; Tissue Distribution

Monoclonal antibodies and lectins were used to examine the expression patterns of apical membrane oligosaccharide sequences specific to type II pneumocytes in atypical adenomatous hyperplasia (AAH) and lung cancer. Atypical cells of AAH and papillary adenocarcinoma cells expressed abundant sialyl Thomsen-Friedenreich (TF) antigen: this was not observed in acinar adenocarcinoma, bronchioloalveolar carcinoma with mucin production or squamous cell carcinoma. Sialyl Tn antigens was also detected on a few cells in AAH and papillary adenocarcinomas. Asialo TF and Tn antigen were not observed on the surface of carcinoma cells of any type. Alpha(alpha)2,3-linked sialic acids predominated in type II pneumocyte, AAH and papillary adenocarcinoma, whereas ciliated columnar cells expressed alpha2,6-linked sialic acids. Lewisx and sialyl Lewisx antigens capped the TF antigen in both O- and N-linked side chains on the surface of AAH and papillary adenocarcinoma cells, but were not expressed by type II pneumocytes. The findings demonstrate that papillary adenocarcinoma cells resemble type II pneumocytes in that they express abundant sialyl TF surface antigen, but they also express TF-related antigens not found in type II pneumocytes. Apical surface glycoconjugates of AAH have structural characteristics shared by both type II pneumocytes and papillary adenocarcinoma cells.

Topics: Adenocarcinoma; Antigens, Tumor-Associated, Carbohydrate; Glycoconjugates; Humans; Lung; Lung Neoplasms; Oligosaccharides; Sialyl Lewis X Antigen

The prognostic value of the immunohistochemical expression of Sialyl Tn antigen (STn) was evaluated in 242 patients with gastric carcinoma. Formalin-fixed, paraffin-embedded specimens of gastric adenocarcinomas were stained with the monoclonal antibody C1282, produced by immunization with ovine submaxillary mucin (OSM). Positive immunoreactivity for STn was observed in 149 (62%) patients. The expression of STn did not correlate with stage of disease (TNM), tumour location, presence of lymph-node or distant metastases, histological type, age or gender. STn immunoreactivity correlated strongly with overall survival in univariate analysis. The median survival in the STn-positive group was 21 months, in comparison to 38 months in the STn-negative group. The difference in survival between STn-negative and STn-positive tumours was significant in patients with stage-I cancer, but not in patients with stage-II -III or -IV disease. STn immunoreactivity emerged as an independent prognostic factor in Cox multivariate analysis. It is concluded that the immunohistochemical expression of STn is a good marker in the prediction of survival in patients with stage-I gastric carcinoma.

Topics: Adenocarcinoma; Adult; Aged; Aged, 80 and over; Antigens, Tumor-Associated, Carbohydrate; Biomarkers, Tumor; Female; Humans; Immunohistochemistry; Male; Middle Aged; Predictive Value of Tests; Prognosis; Stomach Neoplasms

To determine whether the monoclonal antibody (MoAb) 83D4, previously shown to be highly specific for carcinoma cells, can be used as an immunocytological marker to discriminate between benign and malignant cells in serous effusions; and to test for a correlation between expression of the antigen reacting with MoAb 83D4 on effusion cells and the amount of soluble 83D4 antigen in effusion fluids.. Thirty three pleural and 23 peritoneal effusions from 56 cancer patients with metastatic disease were tested for the presence of Tn associated 83D4 antigen by immunocytochemical staining, and for the presence of soluble antigen in supernatants. The patients had undergone various chemotherapy and radiation therapy protocols.. As a result of the various types of treatment, the cytological characteristics of the cells were often modified and the antigenic epitopes may have been altered. Positive staining for 83D4 MoAb was obtained in 36 (97%) of the 37 malignant effusions, eight (73%) of 11 suspect effusions, and three (38%) of the eight apparently benign effusions (free of malignant cells). In these latter cases, cytological reassessment showed a few suspect cells in two cases. 83D4 soluble antigen was detected in 30 of 37 malignant effusions (81%), five of 11 suspected infusions (46%), and five of eight apparently benign effusions (63%).. Immunocytochemical staining with anti-83D4 antibody is useful for differentiating reactive or atypical mesothelial cells from epithelial cells, especially in breast cancer effusions.

Topics: Adenocarcinoma; Antibodies, Monoclonal; Antigens, Tumor-Associated, Carbohydrate; Ascitic Fluid; Biomarkers, Tumor; Breast Neoplasms; Female; Humans; Ovarian Neoplasms; Pleural Effusion, Malignant; Solubility

Pathogenetic aspects of pancarcinoma T/Tn autoantigens were investigated; they are present in approximately 90% of all carcinomas from incipience and throughout. T/Tn are occluded in noncarcinoma (non-CA) diseased and healthy tissues. By serological and immunohistochemical methods, we found that well-differentiated carcinomata express a higher proportion of T than Tn, while in poorly differentiated carcinomata, Tn predominates over T. Tn density of primary carcinomas correlates positively with aggressiveness, early clinical relapse, and early death. Delayed-type skin hypersensitivity to T (DTHR-T) and solid-phase anti-T antibody immunoassay (SPIA-T), respectively, detected 85% of 461 and 88% of 222 carcinoma patients; < 7% of over 450 benign diseased and healthy subjects reacted positively in these assays. T/anti-T assays are highly effective in detecting incipient (TisN0M0 and T1N0M0) carcinomas: DTHR-T-85% of 41, and SPIA-T-96% of 26 patients. Positive anti-T tests predicted CA in 74% of 47 patients months to years before their biopsy/X-ray turned positive.

Topics: Adenocarcinoma; Antibodies; Antigens, Neoplasm; Antigens, Tumor-Associated, Carbohydrate; Autoantigens; Breast Neoplasms; Carcinoma; Carcinoma, Ductal, Breast; Case-Control Studies; Cohort Studies; Humans; Immunoglobulin M; Isoantigens; Prognosis

Alterations in the normal glycosylation process are often associated with oncogenic transformation. Using an anti-Tn monoclonal antibody, MLS128, we have investigated the immunohistochemical localization of Tn antigen in normal and malignant tissues of the digestive tract. In normal tissues, MLS128 was immunoreactive with the squamous epithelium of the esophagus and was weakly reactive with the columnar epithelia of the stomach, duodenum, colon, bile duct and pancreatic duct. In malignant tissues, positive immunostaining was detected with high frequency (75%-100%) in carcinomas of the esophagus, stomach colon, biliary tract and pancreas, whereas 2 of 11 (18%) hepatocellular carcinomas were positive. Tn antigen was detected in the upper two-thirds of the normal squamous epithelium, and was often detected in squamous cell carcinomas with cancer pearls (keratinization). These results suggest that the expression of Tn antigen is related to the differentiation of squamous epithelium, or to keratinization. In normal columnar epithelial cells. Tn antigen was localized mainly to the Golgi area. This intracellular localization was preserved in well-differentiated papillary adenocarcinomas of the colon, but was lost in most cases of tubular adenocarcinomas.

Topics: Adenocarcinoma; Antibodies, Monoclonal; Antibody Specificity; Antigens, Tumor-Associated, Carbohydrate; Carcinoma, Squamous Cell; Digestive System; Digestive System Neoplasms; Epithelium; Humans; Immunohistochemistry

The Tn determinant (GalNAc alpha-O-Ser/Thr) is expressed by about 90% of human carcinomas, but is cryptic in most normal human tissues. A murine monoclonal antibody (MAb) 83D4, developed following immunization with human breast carcinoma cells, reacts with a Tn-related epitope. In the present study we characterized the glycoprotein antigen identified by 83D4 in the human breast carcinoma cell line MCF-7. We further showed that the 83D4 antigenic determinant is masked in human milk fat globule membranes (HMFGM), and can be exposed upon mild m-periodate treatment after desialylation. Western-blot analysis resolved the 83D4 antigen from MCF-7 into two main components of 120-190 kD and > 500 kD respectively. Non equilibrium pH gradient electrophoresis/SDS PAGE revealed the acidic nature of the reactive glycoproteins (pI 4.43-4.70). 83D4 antigenic activity resolved by CsCl gradient ultracentrifugation layered on a wide range of densities (1.30-1.46 g/ml) including typical densities of mucin-like glycoproteins but also lower densities. The amino acid composition of the antigen, relatively rich in serine but poor in threonine and proline, confirmed the divergence from other mucin-like carcinoma-associated glycoproteins. Dicarboxylic amino acids were abundant, accounting in part for the acidic nature of the molecules. ELISA and Western-blot analysis of the subcellular fractions from MCF-7 cells revealed that the 83D4 antigen is mainly contained in plasma membranes (85%) from which it may be resolved into two broad bands (slow and fast migrating components). These results provide information on a group of breast carcinoma associated glycoproteins related to but different from typical mucins, and provide data on alteration of O-glycosylation in tumor cells.

Topics: Adenocarcinoma; Amino Acids; Animals; Antibodies, Monoclonal; Antigens, Neoplasm; Antigens, Tumor-Associated, Carbohydrate; Breast Neoplasms; Carbohydrate Sequence; Epitopes; Female; Glycoproteins; Glycosylation; Humans; Lectins; Membrane Glycoproteins; Mice; Milk, Human; Molecular Sequence Data; Molecular Weight; Mucin-1; Mucins; Neoplasm Proteins; Neuraminidase; Protein Binding; Protein Processing, Post-Translational; Subcellular Fractions; Tumor Cells, Cultured

The aim of this study was to evaluate the expression of blood group Tn and sialyl-Tn antigens in the pancreas to determine whether they could help to interpret histochemically needle biopsies obtained from the pancreas. Lectin and immunohistochemistry was carried out using the biotin-labeled Vicia villosa agglutinin isolectin B4 and the mouse monoclonal antibody MLS102 to detect the Tn and sialyl-Tn blood-group antigens in the pancreas. All the pancreatic ductal adenocarcinomas (11/11) were positively stained by V. villosa agglutinin and MLS102 monoclonal antibody. None of the normals or chronic pancreatitics bound MLS102 monoclonal antibody. The acini of all the normals and chronic pancreatitics were V. villosa agglutinin positive, which was absent in the normal ductal cells and present only sparingly in the chronic pancreatitis ductal tissues (10/16). Thus, both the Tn and the sialyl-Tn blood-group antigens are present in pancreatic ductal tissues that have undergone malignant transformation. MLS102 is superior to V. villosa agglutinin in distinguishing malignant from normal and nonmalignant pancreatic tissues in needle biopsies.

Topics: Adenocarcinoma; Antibodies, Monoclonal; Antigens, Tumor-Associated, Carbohydrate; Biomarkers, Tumor; Biopsy, Needle; Chronic Disease; Humans; Immunohistochemistry; Lectins; Pancreas; Pancreatic Neoplasms; Pancreatitis; Plant Lectins

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.

Topics: Adenocarcinoma; Antigens, Neoplasm; Antigens, Tumor-Associated, Carbohydrate; Biomarkers, Tumor; Cystadenoma; Female; Humans; Immunoenzyme Techniques; Mucins; Ovarian Neoplasms

In a previous report we suggested that T antigen appeared to be associated with gastric carcinoma. To verify this hypothesis and characterize the pattern of expression of simple-mucin type carbohydrate antigens (Tn,sialyl-Tn and T before and after neuraminidase) in normal gastric mucosa and precursor lesions of gastric carcinoma, we studied the mucosa adjacent to 100 cases of gastric carcinoma, gastric biopsies of 60 dyspeptic patients, eight adenomatous polyps and eight hyperplastic polyps. The expression of the antigens was more related to the cell type and underlying lesions than to the coexistence of carcinoma. The most distinctive findings concerned intestinal metaplasia, dysplasia and hyperplastic lesions. In intestinal metaplasia, Tn was found mostly in columnar cells and sialyl-Tn in goblet cells. T was more prevalent in incomplete intestinal metaplasia than in complete. A high prevalence of sialyl-Tn expression and cell membrane immunoreactivity for T antigen, similar to those previously found in gastric carcinomas, were observed in three adenomatous polyps, one hyperplastic polyp, five cases of adenomatous dysplasia in the neighbourhood of intestinal carcinomas and four cases of marked foveolar hyperplasia, three of which were from the mucosa adjacent to diffuse carcinomas. We conclude that adenomatous and hyperplastic lesions share with gastric carcinomas features of aberrant glycosylation, namely the cell membrane expression of T antigen.

Topics: Adenocarcinoma; Adenomatous Polyps; Antigens, Tumor-Associated, Carbohydrate; Carbohydrate Sequence; Dyspepsia; Gastric Mucosa; Gastritis; Gastritis, Atrophic; Glycosylation; Humans; Hyperplasia; Immunoenzyme Techniques; Metaplasia; Molecular Sequence Data; Polyps; Precancerous Conditions; Stomach Neoplasms

The significance of altered expression of MN blood group antigens was examined by studies on the expressions of Thomsen-Friedenreich antigen (T antigen) and Tn antigen in primary and metastatic lesions of 29 human uterine cervical cancers. These antigens were measured by the avidin-biotin-peroxidase (ABC) method with peanut agglutinin (PNA) lectin for T antigen and Vicia villosa agglutinin (VVA) lectin for Tn antigen. Proportion of cancer cells expressing Tn antigen was higher in the metastatic lesions than in the primary tumors in 10 of the 29 cases, less in the metastasis than in the primary tumor in one case, and similar in the primary and metastatic lesions in the other 18 cases. Reaction for Tn antigen was positive in 24 (82.8%) of the 29 metastases, and in 17 (58.5%) of the 29 primary lesions. Thus, the rate of Tn antigen expression was significantly higher in the metastases than in the primary lesions (P < 0.05). On the other hand, there was no significant difference between the immunoreactivities of T antigen in metastases and primary tumors. These findings support our previous suggestion that expression of Tn antigen is closely related to the metastasis to regional lymph nodes and may reflect an important role of this carbohydrate in the process of metastasis of cervical cancer.

Topics: Adenocarcinoma; Antigens, Neoplasm; Antigens, Tumor-Associated, Carbohydrate; Carcinoma, Squamous Cell; Female; Humans; Lymphatic Metastasis; Uterine Cervical Neoplasms

Two monoclonal antibodies (MoAbs), BRIC 66 (IgM) and BRIC 111 (IgG1), were produced by immunizing mice with ovarian cyst blood group A1 glycoprotein and Tn red cells (RBCs), respectively. Their specificities were determined by inhibitions using Tn sialoglycoproteins (SGPs), mucins (armadillo [ASG] and ovine [OSG] submaxillary glycoproteins), and monosaccharides. BRIC 66 agglutinated both Tn and group A RBCs and reacted immunohistochemically with both the vascular endothelium and tumor cells from a group A adenocarcinoma, BRIC 66 was inhibited by N-acetylgalactosamine (GalNAc), Tn SGPs, and mucins on both hemagglutination inhibition tests and radioimmunoassay. BRIC 111 agglutinated Tn RBCs only, and it specifically stained tumor cells from a group O patient's breast carcinoma and a group A patient's adenocarcinoma. In hemagglutination inhibition tests, BRIC 111 was readily inhibited by Tn SGPs, only partially inhibited by GalNAc, and not inhibited by mucins. In a sensitive radioimmunoassay, BRIC 111 was inhibitable by GalNAc. Tn SGP was 2000-fold more effective as an inhibitor than the mucins (ASG and desialized OSG), which contain a high content of terminal alpha-GalNAc-O-serine (threonine) residues. It is postulated that BRIC 66 is specific for terminal alpha-GalNAc units in carbohydrate chains. The exclusive reaction of BRIC 111 with Tn SGP indicates a combining site larger than GalNAc alpha-1, which probably includes amino acid residues in juxtaposition to GalNAc in Tn SGP. In view of its specific agglutination of Tn RBCs, BRIC 111 is a useful reagent for the examination of polyagglutinable RBCs.

Topics: ABO Blood-Group System; Absorption; Acetylgalactosamine; Adenocarcinoma; Antibodies, Monoclonal; Antigens, Neoplasm; Antigens, Tumor-Associated, Carbohydrate; Breast Neoplasms; Erythrocytes; Female; Glycoproteins; Hemagglutination; Humans; Immunoenzyme Techniques; Mucins; N-Acetylneuraminic Acid; Neuraminidase; Ovarian Cysts; Oxidation-Reduction; Periodic Acid; Sialic Acids; Sialoglycoproteins

Carbohydrate antigens representing some of the initial steps in mucin O-linked glycosylation were examined in specimens of normal pancreas, chronic pancreatitis, and pancreatic adenocarcinoma. Tn antigen, recognized by Vicia villosa lectin, was expressed by all specimens of normal pancreas (acinar cells) and pancreatic cancers and all but one case of chronic pancreatitis. Sialosyl Tn antigen, recognized by monoclonal antibody TKH2, was expressed in a cancer-associated fashion, being completely absent in normal pancreas but expressed by 56% of chronic pancreatitis and 97% of pancreatic cancers. T antigen, recognized by monoclonal antibody AH9-16, was expressed in 68% of normal pancreas (acinar cells), 67% of chronic pancreatitis, and 48% of pancreatic cancer tissues. These results indicate that normal acinar cells of the pancreas are capable of expressing selected carbohydrate structures associated with the initial steps of mucin glycosylation. The marked expression of sialosyl Tn compared with T antigen in pancreatic cancers suggests that with malignant transformation there is selective usage of glycosyltransferase enzymes involved in mucin oligosaccharide synthesis.

Topics: Adenocarcinoma; Adenocarcinoma, Mucinous; Antibodies, Monoclonal; Antigens, Neoplasm; Antigens, Tumor-Associated, Carbohydrate; Biomarkers, Tumor; Chronic Disease; Disaccharides; Humans; Immunoenzyme Techniques; Pancreas; Pancreatic Neoplasms; Pancreatitis

Nine cases of malignant mesothelioma of pure epithelial and biphasic types (five pleural, three peritoneal, and one pericardial mesotheliomas), seven cases of benign adenomatoid tumor of the uterus, and 21 cases of peripheral pulmonary adenocarcinoma of non-mucus-producing type were examined immunohistochemically for expression of keratin, vimentin, carcinoembryonic antigen (CEA), surfactant apoprotein, Lewis blood group antigens, and Tn antigen. The majority (78%) of the malignant mesotheliomas expressed keratin, but CEA and surfactant apoprotein were not detected in any mesotheliomas. On the other hand, pulmonary adenocarcinomas expressed not only keratin (100%), but also CEA (62%) and surfactant apoprotein (62%). The expression of Lewisa blood group antigen and Tn antigen was detected in 76% and 62% of the pulmonary adenocarcinomas, respectively, but only one mesothelioma was stained for Lewisa antigen. This study reveals that the majority of malignant mesotheliomas can be distinguished from pulmonary adenocarcinomas by immunohistochemcial staining for CEA, surfactant apoprotein, Lewisa antigen, and Tn antigen. Immunohistochemically, adenomatoid tumors behaved similarly to malignant mesotheliomas.

Topics: Adenocarcinoma; Antibodies, Monoclonal; Antigens; Antigens, Neoplasm; Antigens, Tumor-Associated, Carbohydrate; Carcinoembryonic Antigen; Diagnosis, Differential; Histocytochemistry; Humans; Immunohistochemistry; Intermediate Filament Proteins; Isoantigens; Lewis Blood Group Antigens; Lung Neoplasms; Mesothelioma

The Thomsen, Friedenreich (TF) and Tn carbohydrate antigens are expressed on the vast majority of human adenocarcinomas and are associated with aggressive behavior of certain tumors. TF and Tn antigens are also expressed on certain murine cancer cell lines including TA3-Ha, a highly lethal, transplantable mammary adenocarcinoma. TF and Tn cancer-associated carbohydrate haptens were synthesized, conjugated to protein carriers and used to demonstrate that delayed-type hypersensitivity (DTH) effector T cells can specifically recognize and respond to carbohydrate determinants on the TA3-Ha tumor-associated glycoprotein, epiglycanin. The effector cells were shown to have the helper DTH phenotype (Lyt1+, Lyt2-, Thy1+) and it was demonstrated that they respond to specific carbohydrate determinants in an MHC-restricted fashion. These experiments provide the rationale for the use of synthetic tumor-associated glycoconjugates (S-TAGs) to stimulate anticancer T cell immunity. In support of this hypothesis, it was shown that preimmunization with the appropriate S-TAGs could provide a degree of protection against a subsequent tumor transplant and that antitumor effector Lyt1+, Lyt2- T cells could be generated in vitro using the appropriate S-TAGs as antigens.

Topics: Adenocarcinoma; Animals; Antigens, Neoplasm; Antigens, Tumor-Associated, Carbohydrate; Carbohydrates; Disaccharides; Epitopes; Hypersensitivity, Delayed; Immunization; Immunization, Passive; Macrophage Activation; Mammary Neoplasms, Experimental; Membrane Glycoproteins; Mice; Mice, Inbred Strains; Neoplasm Proteins; Neoplasm Transplantation; T-Lymphocytes, Cytotoxic

Topics: Adenocarcinoma; Agglutination Tests; Antibodies; Antibody Specificity; Antigens, Neoplasm; Antigens, Tumor-Associated, Carbohydrate; Breast Neoplasms; Carcinoma; Carcinoma, Intraductal, Noninfiltrating; Cell Line; Cell Migration Inhibition; Cytotoxicity, Immunologic; Disaccharides; Epitopes; Erythrocytes; Female; Humans; Hypersensitivity, Delayed; Killer Cells, Natural; Leukocytes; Skin Tests