morphine and Adenocarcinoma

To obtain monoclonal antibodies directed to mucin-type sugar chains, mice were immunized with bovine submaxillary mucin (BSM) that had been conjugated with ovalbumin. Conjugation of BSM with ovalbumin enhanced the antigenicity of BSM to about five to ten times that of intact BSM and resulted in the establishment of ten hybridomas, all of which secreted monoclonal antibodies toward BSM. Most of the antibodies secreted by these hybridomas did not react with glycolipids but did react with glycoproteins. Several antibodies lost their reactivity when sialic acid residues were removed from BSM, indicating that these antibodies recognize carbohydrate moieties of mucins. Immunohistochemical studies revealed that three of the antibodies recognized human ovarian cancer-associated carbohydrate antigens. In addition, one of these three antibodies reacted with a human cultured colonic cancer cell line. The protocol described in this paper was effective in producing monoclonal antibodies that recognize mucin-carbohydrates and some of the generated antibodies can be applied to the detection of cancers.

Topics: Adenocarcinoma; Animals; Antibodies, Monoclonal; Antigens, Neoplasm; Brain Chemistry; Cattle; Cell Line; Colorectal Neoplasms; Enzyme-Linked Immunosorbent Assay; Glycolipids; Glycoproteins; Humans; Meconium; Mice; Mucins; Rabbits; Sheep; Submandibular Gland; Tumor Cells, Cultured

Reactivity of neuraminidase-treated colorectal carcinoma cells with antibodies that detect the X-carbohydrate structure was greater than the reactivity of untreated cells. The same results were obtained with glycolipid extracts of meconium, a colorectal carcinoma cell line, three freshly excised human adenocarcinomas, and normal bronchial mucosa. The glycolipid was either not expressed or expressed in smaller quantities on the corresponding normal colon tissue. Further study showed that the major sialo-X glycolipid has six sugars including a single sialic acid which blocks X-antigenicity. These glycolipids were further analyzed by ion-exchange high-pressure liquid chromatography and thin-layer chromatography. These monosialo-X glycolipid antigens might serve as potential tumor markers.

Topics: Adenocarcinoma; Antibodies, Monoclonal; Antigens, Neoplasm; Antigens, Surface; Bronchi; Cell Line; Chromatography, High Pressure Liquid; Chromatography, Thin Layer; Colonic Neoplasms; Fetal Proteins; Glycolipids; Humans; Infant, Newborn; Meconium; Mucous Membrane; Neuraminidase; Radioimmunoassay

Two antigens cross-reactive with carcinoembryonic antigen (CEA) and distinct from the nonspecific cross-reacting antigen were identified in meconium by double immunodiffusion with a conventional goat anti-CEA antiserum. These two antigens together competitively inhibited cross-reacting antibodies against them in CEA radioimmunoassay and contributed to the measurement of meconium CEA levels which averaged 6 times higher than that determined with anti-CEA specific antibody. A purification method for one of these antigens, tentatively designated meconium antigen, is described and uses a combination of ethanol fractionation, ion-exchange and molecular sieve chromatography, and adsorption to an immunoadsorbent containing a cross-reactive murine monoclonal antibody to CEA. Preliminary characterization of the purified meconium antigen showed it to be a glycoprotein, migrating as an alpha-globulin and having a molecular size similar to that of CEA (Mr 185,000 versus 200,000). Antigenically, it lacks at least one determinant present on CEA and differs further from CEA by being weakly reactive with concanavalin A and resistant to proteolytic digestion with Pronase E. Although these properties of meconium antigen suggest that it may be nonspecific cross-reacting antigen 2, additional chemical and antigenic studies are required to establish its relationship to CEA and other CEA-related antigens in meconium.

Topics: Adenocarcinoma; Antigens; Carcinoembryonic Antigen; Colonic Neoplasms; Humans; Immunodiffusion; Infant, Newborn; Liver Neoplasms; Meconium; Radioimmunoassay