Carcinoembryonic antigen-identification of a tri-N-acetylchitotriosyl type structure as an immunodeterminant group

Anderson, Byron; Jameson, Alice K.; Hirata, Arthur A.; Safford, J W; Tomita, Joseph T.

doi:10.1016/0019-2791(75)90088-9

Cited by 9 publications

(3 citation statements)

References 15 publications

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“…Regarding the chemical nature of the various antigenic determinants of the CEA molecule, there are several conflicting reports. As immunoactive carbo-hydrate moiety, glycopeptides with molecular weights of 1,000-5,000 (Banjo et al, 1974), and tri-N-acetylchitotriose, a tri-saccharide of ,81+4 linked N-acetylglucosamine (Anderson et al, 1975) have been proposed. On the other hand, it has been reported by others (Hammarstrom et al, 1975;Westwood and Thomas, 1975) that the carbohydrate moiety of CEA does not contain the tumorassociated determinant(s).…”

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confidence: 99%

Proteolytic release of antigenic fragments corresponding to normal fecal antigen and non‐specific cross‐reacting antigen from carcinoembryonic antigen

Matsuoka

Koga

Maruta

et al. 1978

Intl Journal of Cancer

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Three immunogenic parts have so far been identified in the carcinoembryonic antigen (CEA) molecule. These are: determinants cross-reactint with the normal fecal antigen (NFA) (NFA determinant); determinants cross-reacting antigen (NCA) (NCA determinant); and determinants which appear to be more cancer-specific (cancer determinant). The chemical nature of these parts of the CEA molecule was investigated by digestion with proteolytic enzymes together with anti-CEA preparations with which these three immunogenic parts of CEA molecule could be identified. The CEA digest obtained with pepsin did not react in immunodiffusion and radioimmunoassay, indicating that pepsin completely destroyed all the antigenic parts. Digestion by pronase E destroyed only the cancer determinant and liberated two antigenic fragments corresponding to the NFA determinant and the NCA determinant, respectively. These results suggest that the cancer determinant may reside in a protein or a peptide part of the molecule. The chemical nature of the NFA and NCA determinant remains to be clarified.

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confidence: 99%

Proteolytic release of antigenic fragments corresponding to normal fecal antigen and non‐specific cross‐reacting antigen from carcinoembryonic antigen

Matsuoka

Koga

Maruta

et al. 1978

Intl Journal of Cancer

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“…Wheat germ agglutinin reacts very well with CEA (Goldstein et al, 1975;Hammarstrom et al, 1975), while Aaptos lectin I reacts weakly. Since periodate oxidation and methylation studies (Hammarstrom et al, 1975;Fuks et al, 1975) indicate the absence of terminal nonreducing dGIcNAc in CEA, the reactive dGIcNAc must be internal and the findings of Anderson et al (1975) that AA^A'^triacetylchitotriose was better than the dimer or p-nitrophenyl-8dG1cNAc could indicate specificity for an internal sequence of one or more 8-linked dGIcNAc residues. Thus the poor reaction of CEA with Aaptos lectin I supports the inference that the Aaptos lectin I site is directed against terminal nonreducing chains of 81-4 linked dGIcNAc.…”

Section: Discussionmentioning

confidence: 99%

Purification and characterization of the agglutinins from the sponge Aaptos papillata and a study of their combining sites

Bretting¹,

Kabat²,

Liao³

et al. 1976

Biochemistry

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The lectins from the sponge Aaptos papillata were isolated by affinity chromatography using polyleucyl blood group A + H substances from hog stomach linings as an absorbent and eluting with 3 M MgCl2. Further separation on diethylaminoethylcellulose and preparative disc electrophoresis on polyacrylamide gave the three fractions, Aaptos lectins I, II, and III. They were essentially homogenous in polyacrylamide electrophoresis and sedimentation analysis: a small second component was seen in lectins I and II in immunoelectrophoresis at high concentration. The S20,W0 values for Aaptos lectins I, II, and III were 3.5, 6.0, and 5.5. By electrophoresis in sodium dodecyl sulfate with an without beta-mercaptoethanol Aaptos lectin I showed two bands corresponding to molecular weights of 12 000 and 21 000; Aaptos lectins II and III gave only one band of molecular weight of 16 000. In isoelectric focusing, Aaptos lectin I showed bands at pH 4.7 and 5.4 and in the range between 6.8 and 7.6, while Aaptos lectins II and III were almost identical with bands at pH 3.8, 4.7 to 4.9, and 5.3. Aaptos lectin I differed from II and III in amino acid composition but the latter two were very similar. They contained no significant carbohydrate. Aaptos lectin I reacted best with blood group substances with terminal nonreducing N-acetyl-D-glucosamine residues precipitating about two-thirds of the lectin N added while blood group substances with terminal nonreducing DGalNAc were almost inactive. However, Aaptos lectin II was completely precipitated by blood group substances and glycoproteins containing terminal DGalNAc, DGlcNAc, or sialic acid residues. Aaptos lectin III had a precipitation pattern similar to Aaptos lectin II. DGlcNAc but not DGalNAc inhibited precipitation of Aaptos lectin I by blood group substances and N, N', N'', N'''-tetraacetylchitotetraose was the best inhibitor and was 2000 times more active than DGlcNAc. Precipitin reactions with Aaptos lectin II were inhibited by equal amounts of DGlcNAc and by sialic acid which were four times more potent than DGalNAc. N,N',N''-triacetylchiotriose was the best inhibitor and was 13 times better than DGlcNAc. At 37 degrees C three to four times higher amounts of inhibitor were necessary to inhibit precipitation of Aaptos lectin II than were needed at 4 degrees C, indicating higher affinity of blood group substances for Aaptos lectin II with increasing temperature. Aaptos lectin I was precipitated by the monofunctional hapten p-nitrophenyl-alphaDGalNAc, while p-nitrophenyl-betaDGalNAc did not precipitate and was a good inhibitor. Both phenomena indicate involvement of hydrophobic bonds.

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“…(ii) It is one of the few oligosaccharides able to inhibit Abbreviations: WGA, wheat germ agglutinin; StA, Solanum tuberosum agglutinin; CB, di-N-acetylchitobiose; ACBA-L, antibodies against di-N-acetylchitobiose that bind to N-acetylglucosamine; ACBA-2, antibodies against di-N-acetylchitobiose that bind to di-N-acetylchitobiose; BIgG, bovine immunoglobulin; BSA, bovine serum albumin; CB4-BSA, (di-N-acetylchitobiose)4-bovine serum albumin; CBg-BIgG, (di-N-acetylchitobiose)g-bovine IgG cell agglutination induced by wheat germ lectin (WGA), (2)(3)(4); it is known that several tumor cell lines were highly agglutinable by WGA but nontransformed cells were not (2,5). (iii) In some carbohydrate inhibition experiments, it was found that GlcNAc and CB were the immunodominant group of carcinoembryonic antigen (6)(7)(8), however, opposite results were found by other authors (9,10). (iv) CB glycosidically linked to poly-L-aspartate via N-asparaginyl bonds was shown to elicit an immune response which increased the survival times of mice challenged with an intraperitoneal injection of tumor cells (11).…”

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Protein-sugar interactions: preparation, purification, and properties of rabbit antibodies against di-N-acetylchitobiose.

Kiéda¹,

Delmotte²,

Monsigny³

1977

Proc. Natl. Acad. Sci. U.S.A.

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Antibodies against di-N-acetylchitobiose (CB) were raised in rabbits after injection of CB-bovine serum albumin conjugates and were fractionated by columns into two classes: the first bound to a column of Sepharose covalently coupled with N-acetylglucosamine (GlcNAc); the second bound to a column coupled with CB. Active antibodies were eluted by a moderate concentration of a chaotropic agent, but not by high ionic strength buffers or acidic buffers. The active fractions were identified as IgG by ultracentrifugation and immunoelectrophoresis. These antibodies gave precipitation bands with CB-protein conjugates and this reaction could be reversed in the presence of free CB. The GlcNAc-bound fraction quantitatively quenched the fluorescence of 04-methylumbelliferyl) glycosides of GlcNAc and CB, while the CB-bound fraction quenched only the glycoside of CB; other 04-methylumbelliferyl-glycosides were not quenched. Among eleven monosaccharides and oligosaccharides, only GlcNAc, CB, and tri-Nacetylchitotriose were able to inhibit the precipitation of antibodies against CB with CB-protein conjugate. These antibodies failed to agglutinate erythrocytes from various species but did agglutinate transformed cells and mouse lymphocytes. The binding of these antibodies on cell membranes was reversed by free CB and by CB-protein conjugates. The properties of these antibodies are related to those of lectins with similar specificities and to the structure of glycoconjugates. cell agglutination induced by wheat germ lectin (WGA), (2-4); it is known that several tumor cell lines were highly agglutinable by WGA but nontransformed cells were not (2, 5). (iii) In some carbohydrate inhibition experiments, it was found that GlcNAc and CB were the immunodominant group of carcinoembryonic antigen (6-8), however, opposite results were found by other authors (9, 10). (iv) CB glycosidically linked to poly-L-aspartate via N-asparaginyl bonds was shown to elicit an immune response which increased the survival times of mice challenged with an intraperitoneal injection of tumor cells (11). Di-N-acetylchitobiose (CBTo detect and to study the functions of cell surface glycoconjugates, lectins have been extensively used (12, 13). However, on one hand, lectins are usually tetravalent as WGA is (14-16) and therefore lectins agglutinate cells or crosslink the cell surface glycoconjugate; on the other hand, lectins bind large oligosaccharides and glycoproteins more strongly than monosaccharides or small oligosaccharides (13). Recently, carbohydrate specific antibodies were purified from the sera of various animal species by affinity chromatography on immobilized fetuin as an adsorbent (17); the binding of carbohydrate specific antibodies to mouse splenocytes was partially inhibited by various free sugars and by several oligosaccharides. Antibodies raised against some specific oligosaccharides should be a sugar-binding protein with narrow specificity, and Fab fragments easily obtained by proteolytic hydrolysis of the antibodies should be...

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Carcinoembryonic antigen-identification of a tri-N-acetylchitotriosyl type structure as an immunodeterminant group

Cited by 9 publications

References 15 publications

Proteolytic release of antigenic fragments corresponding to normal fecal antigen and non‐specific cross‐reacting antigen from carcinoembryonic antigen

Proteolytic release of antigenic fragments corresponding to normal fecal antigen and non‐specific cross‐reacting antigen from carcinoembryonic antigen

Purification and characterization of the agglutinins from the sponge Aaptos papillata and a study of their combining sites

Protein-sugar interactions: preparation, purification, and properties of rabbit antibodies against di-N-acetylchitobiose.

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