Lewisx Antigen Research Articles

Immunohistochemical analysis of the poorly differentiated stomach adenocarcinoma with medullary growth pattern

Poorly differentiated gastric adenocarcinoma with medullary features (poor medullary) is distinguished by a propensity for hepatic metastasis. To classify it antigenically, we compared it to poorly differentiated adenocarcinoma with scirrhous growth pattern (poor scirrhous), well and moderately differentiated adenocarcinoma (differentiated adenocarcinoma), and normal gastric mucosa (foveolar and deep epithelium) using immunohistochemistry with antibodies against CEA, AFP, NSE, and Lewis-type antigens. Lewisb antigen was significantly associated with differentiated adenocarcinoma and foveolar epithelium, although Lewisx antigen was significantly expressed in poor medullary, poor scirrhous, and deep gland epithelium. From the viewpoint of expression of Lewisa, there was no significant differentiation between poor medullary and differentiated adenocarcinoma, but it was definite between poor scirrhous and differentiated adenocarcinoma. Therefore, we conclude that in antigenic expression, poor medullary carcinoma is allied with differentiated adenocarcinoma rather than poorly differentiated scirrhous carcinoma. © 1996 Wiley-Liss, Inc.

Characterization of Human Colon Carcinoma Variant Cells Selected for Sialyl Lex Carbohydrate Antigen: Liver Colonization and Adhesion to Vascular Endothelial Cells

The content of sialyl Lewis-X antigen (Neu5Acα2-3Galβ1-4(Fucα1-3)GlcNAc-R: sialyl Lex) was previously shown to correlate with the progression of human colon carcinomas to the advanced stages. Variant cell lines with high (KM12-HX) or low (KM12-LX) levels of cell surface sialyl Lex were isolated from the heterogeneous KM12C cells to characterize the biological behavior of colon carcinoma cells with elevated cell surface contents of sialyl Lex. When these cells were injected intrasplenically into nude mice, KM12-HX cells colonized to the liver more efficiently than KM12-LX cells. Under in vitro conditions, KM12-HX cells demonstrated greater degree of adhesion to cytokine-activated human umbilical vein endothelial cells than KM12-LX cells. The adhesion of KM12-HX cells was partially inhibited by antibodies specific for E-selectin, which was known to serve as a ligand for the sialyl Lex carbohydrate antigen. The treatment of KM12-HX cells with benzyl N -acetyl-α-D-galactosaminide, a putative inhibitor of the extension of O-linked carbohydrate chains, reduced the rate of adhesion. These results suggested that the interaction of endothelial cell surface E-selectin with O-linked carbohydrate chains on colon carcinoma cell surface glycoproteins played an important role in the adhesion.

Profiles of expression of carbohydrate chain structures during human intrahepatic bile duct development and maturation: A lectin-histochemical and immunohistochemical study

We investigated expression of carbohydrate chain structures during human intrahepatic bile duct development and maturation. Results of staining for ABH antigens were negative in the ductal plate and biliary cells migrating into the mesenchyma but were positive in newly formed bile ducts in the fetal liver and in maturing bile ducts in the postnatal liver. Staining for Lewis(a), sialyl Lewis(a) and Lewisb antigens was negative in the ductal plate but positive in migrating biliary cells, newly formed ducts and maturing ducts. Staining for sialyl Lewisx antigen was positive in the ductal plate, migrating biliary cells and newly formed ducts but negative in the postnatal liver. Lewis(y) antigen staining was negative in the ductal plate and migrating biliary cells but positive in newly formed ducts and maturing ducts. Staining for T and Tn antigens was negative in the ductal plate, migrating biliary cells and newly formed ducts but positive in maturing ducts. Staining for Lewisx, sialyl Tn and carcinoembryonic antigen was negative in all tissues. Receptors of Ulex europaeus agglutinin I and soybean agglutinin were absent in the ductal plate, migrating biliary cells and newly formed ducts but present in maturing large ducts. Staining for receptors of concanavalin A and Ricnus communis agglutinin I was negative in the ductal plate but positive in migrating biliary cells, newly formed ducts and maturing ducts. Staining for the receptor of succinylated wheat germ agglutinin was always positive. These findings suggest that carbohydrate chain structures change during intrahepatic bile duct development and maturation and that new carbohydrate chain structures gradually emerge as intrahepatic bile ducts develop and mature.

Cancer-associated quantitative differences in antigen levels of sialosyl Lewisx in stool from patients with colorectal carcinomas

A microenzyme-linked immunosorbent assay system employing monoclonal antibody SNH3 was developed for the detection of sialosyl Lewisx antigen in stool extracts from 80 patients with colorectal cancer, 13 patients with colorectal non-malignant disorders and 90 normal subjects. Sialosyl Lewisx antigen was detected in 35% of stool extracts from cancer patients but only in 7.7% and 2.2% of those from non-malignant patients and normal subjects respectively. Hemoglobin in the same stool samples was detected in 37.5% of cancer patients, 15.4% of non-malignant patients and 5.6% of normal subjects. The appearance of sialosyl Lewisx antigen in stool was not necessary correlated with that of hemoglobin, and overall 61.3% of cancer patients were detected by the combination of the two assays. The combination assay was also impressive in early detection of colorectal cancer (Dukes' A, 52%; Dukes' B, 57.1%). Therefore, the assay for sialosyl Lewisx antigen in stool would be useful for detecting colorectal cancer.

Synthesis of N‐Glycopeptide Clusters with Lewisx Antigen Side Chains and Their Coupling to Carrier Proteins

As a tumor-associated cell-antigen mimic, Lewisx trisaccharide determinant 1 was constructed with the azide function as anomeric protecting group with the aid of a 4-Mpm-protected (Mpm = methoxyphenylmethyl) fucosyl donor. Exchange of the OMpm functions for OAc groups furnishes a Lewisx trisaccharide stable enough against acid attack for glycopeptide synthesis. A glycopeptide with two Lewisx antigen side chains was deblocked and coupled to bovine serum albumin as well as to KLH (keyhole limpet hemocyanin). These synthetic glycoproteins simulate biantennary Lewisx antigen structures.

S4.8 Expression of the Lewisx and sialyl Lewisx antigens on mouse Lewis lung carcinoma LL2 cell Line and its Aleuria aurantia agglutinin-resistant high-metastatic variant

S4.8 Expression of the Lewisx and sialyl Lewisx antigens on mouse Lewis lung carcinoma LL2 cell Line and its Aleuria aurantia agglutinin-resistant high-metastatic variant

CD66 nonspecific cross-reacting antigens are involved in neutrophil adherence to cytokine-activated endothelial cells.

Neutrophil adherence to cytokine-activated endothelial cell (EC) monolayers depends on the expression of the endothelial leukocyte adhesion molecule-1 (ELAM-1). The ligand for ELAM-1 is the sialylated Lewis-x antigen (SLe(x)) structure. The selectin LAM-1 (or LECAM-1) has been described as one of the SLe(x)-presenting glycoproteins involved in neutrophil binding to ELAM-1. Other presenter molecules have not yet been described. Our data demonstrate that the carcinoembryonic antigen (CEA)-like surface molecules on neutrophils--known as the nonspecific cross-reacting antigens (NCAs)--are involved in neutrophil adherence to monolayers of IL-1-beta-activated EC. The NCAs are recognized by CD66 (NCA-160 and NCA-90) and CD67 (NCA-95). Because NCA-95 and NCA-90 have previously been found to be phosphatidylinositol (PI)-linked, paroxysmal nocturnal hemoglobinuria (PNH) neutrophils (which lack PI-linked surface proteins) were tested as well. PNH neutrophils showed a diminished binding to activated EC. CD66 (on PNH cells still recognizing the transmembrane NCA-160 form) still inhibited the adherence of PNH cells to IL-1-beta-activated EC, but to a limited extent. Soluble CEA(-related) antigens inhibited normal neutrophil adherence as well, whereas neutrophil transmigration was unaffected. Sialidase-treatment as well as CD66 preclearing abolished the inhibitory capacity of the CEA(-related) antigens. The binding of soluble CEA antigens to IL-1-beta-pretreated EC was blocked by anti-ELAM-1. These soluble antigens, as well as the neutrophil NCA-160 and NCA-90, both recognized by CD66 antibodies, presented the SLe(x) determinant. Together, these findings indicate that the CD66 antigens (i.e., NCA-160/NCA-90) function as presenter molecules of the SLe(x) oligosaccharide structures on neutrophils that bind to ELAM-1 on EC.

Analysis of the Lewisx epitope in human pancreas and pancreatic adenocarcinomas.

The Lewisx epitope (Gal beta 1-4[Fuc alpha 1-3]GlcNAc-R) can be detected in most ductal pancreatic carcinomas. However, few data pertain to its expression in normal exocrine pancreas and its biochemistry. We compared the expression of the Lewisx epitope in normal pancreas with its expression in pancreatic adenocarcinomas and tumor cell lines and to further characterize the nature of the antigens bearing this epitope with immunochemical methods. On frozen tissue sections of normal pancreas, the Lewisx epitope was detected focally in acinar cell granules; sialosyl-Lewisx was detected in centroacinar cells and the cytoplasm of intralobular ducts. Sixteen of 19 pancreatic carcinomas expressed the Lewisx antigen on tissue sections; however, there was no correlation with prognostic parameters, such as tumor grade or stage. Eighteen of 19 tumor specimens were positive for sialosyl-Lewisx. Analysis of pancreatic carcinoma cell lines (CAPAN-1, CAPAN-2, and DAN-G) revealed intracytoplasmic expression of sialosyl-Lewisx epitopes in these cells, and cell surface reactivity was detected on flow cytometry for sialosyl-Lewisx. Lectin-affinity chromatography of cytoplasmac preparations showed that Lewisx-positive antigens of normal human pancreas bind to SBA and UEA-I lectins but have little or no affinity to WGA, GS-I, or DBA lectins, indicating the presence of Fuc and Gal oligosaccharide residues. It was concluded that the Lewisx and sialosyl-Lewisx antigens are nonpolymorphic carbohydrate determinants that are present in secretory granules of acinar cells, ductules, and pancreatic secretions. This makes the Lewisx antigen suitable for the analysis of the secretory process in the exocrine pancreas and the study of secretory differentiation antigens in ductal pancreatic carcinoma.

SIALYL LEWISx AS A TUMOR-ASSOCIATED CARBOHYDRATE ANTIGEN IN BREAST CANCER

Operative breast cancers were subjected to a serological and immunohistochemical evaluation of sialyl Lewisx antigen as a tumor-associated one, where a new monoclonal antibody CSLEX1 which reacts with sialyl Lewisx antigen was used. Percent positive of sailayl Lewisx in sera of 30 patients with breast cancer was 30%(9/30), which was better than those of any other tumor markers such as CEA, CA19-9, CA15-3 and SLX. Percent positives, according to clinical stages, were 20.0%, 37.5%, 40.0% and 50.0% in stage I, II, III and IV respectively. Immunohistochemically, the expression of a sialyl Lewisx in the normal ductal epithelium and carcinoma cells of 41 breast cancers demonstrated 95.5%(21/22) and 65.9%(27/41) respectively. Although the immunohistochemical experssion of sialyl Lewisx antigen indicated a low cancer specificity, comparatively high positive rates in stage I and II were obtained in sera of cancer patients. It has been suggested that sialyl Lewisx antigen is useful as a tumor-associatd antigen in breast cancers.

Expression of Lewis<sup>X</sup> and Sialylated Lewis<sup>X</sup> Antigens in Human Colorectal Polyps<xref ref-type="fn" rid="FN2">2</xref>

The LewisX (LeX) antigen [characterized by trisaccharide Gal beta 1----4 (Fuc alpha 1----3)N-acetylglucosamine] is an oncodevelopmental antigen in the human colon. Monoclonal antibodies (MoAbs), anti-SSEA-1 and AH8-183, which recognize LeX antigen either on short oligosaccharide side chains or as a terminal immunodeterminant on longer carbohydrate side chains of glycoconjugates, bind to most colon cancer tissues but also to some normal colon mucosae. However, the monoclonal antibodies FH1, FH4, FH6, and IB9, which recognize extended difucosylated and trifucosylated LeX structures or their sialylated derivatives, are more cancer-associated because they rarely bind to normal colon mucosa. In the present study, these MoAbs were used to compare the expression of various LeX-related antigens in premalignant (adenomatous) and nonpremalignant (hyperplastic) colorectal polyps. Antigen expression in polyps was also compared to antigen expressions of normal colon mucosa and colon cancer tissues. The four MoAbs recognizing extended LeX antigens bound to adenomatous polyps (APs) significantly more than to hyperplastic polyps (HPs). In contrast, anti-SSEA-1 and AH8-183 recognizing monofucosyl LeX were less able to distinguish between APs and HPs. In APs, staining with the four MoAbs recognizing extended LeX antigens correlated with the premalignant parameters of larger polyp size, more severe dysplasia, and increased villose component. However, staining with AH8-183 correlated only with polyp size, and anti-SSEA-1 correlated only with polyp size and degree of dysplasia. In general, the staining frequency of HPs was similar to that of normal colon mucosa, although FH6, which did not stain any specimens of normal mucosa, stained a few HPs. The staining frequency of APs was less than that of colon cancer tissues, but these differences were generally not statistically significant. In conclusion, extended LeX antigens and their sialylated derivatives are cancer-associated antigens that are expressed preferentially in premalignant colon polyps, that tend to correlate with malignant potential in these polyps, and that may eventually help to define mechanisms involved in the polyp-to-cancer sequence.