Characterization of Glycolipids from the Gastric

[CANCER RESEARCH 47, 1968-1972, April 1, 1987] Characterization of Glycolipids from the Gastric Cancer of a Patient of p,O,Le(a-,b+) Blood Type: Presence of Incompatible Blood Group Antigens in Tumor Tissues Hiroshi I fattori,1 Kei-ichi Uemura, Hiroyuki Ogata, Tsutomu Katsuyama, Tamotsu Taketomi, and Julian N. Kanfer

Department ofLipid Biochemistry, Institute of Cardiovascular Disease [H. H., K. U., T. T.], Blood Transfusion Service Â¡H.O.J,and Department of Laboratory Medicine IT. K.I, Shinshu University School of Medicine, 3-1-1, Asahi, Matsumoto, Japan 390, and Department of Biochemistry, Faculty of Medicine, University of Manitoba Winnipeg, Manitoba, Canada R3E OW3 Â¡J.K.]

ABSTRACT tissues. The chemical and immunological properties of glyco lipid fractions purified from these tissues were examined. The antigens present in a gastric tumor obtained from a patient of blood group p,O,Le(a-,b+) were studied. The neutral glycolipids were isolated from the cancer tissues and characterized chemically and im- MATERIALS AND METHODS munologically. The glycolipid pattern of the tumor was similar to that of the surrounding uninvolved mucosae. The major neutral glycolipids were Case Report. Mrs. Y. K., a 69-year-old housewife, visited the Ngano found to be glucosylceramide, galactosylceramide, lactosylceramide, and Red Cross Hospital because of weight loss. Physical examination and the lacto series glycolipids, including the H, Le*,and Lebactive fucolipids. a routine chemical and hematological study were essentially negative. The cancer tissues and the uninvolved mucosae did not contain galabio- Roentgenographic examination of her stomach disclosed one ulcerous sylceramide, globotriaosylceramide, or globotetraosylceramide after sep mass at the upper antrum and another ulcerous smaller mass at the arations by both thin layer chromatography and gas-liquid chromatog- upper cardia. A total gastrectomy was scheduled because of the diag raphy. Thin layer chromatography immunostaining was performed to nosis of a double gastric carcinoma and blood was ordered for trans detect incompatible blood group antigens. Immunostaining revealed the fusion. The patient was O, Rh positive. Compatible blood was unob presence of globotriaosylceramide, globotetraosylceramide, Forssman tainable because of an unexpected antibody. The patient underwent glycolipid, and A active glycolipids in the cancer tissues. These incom surgery on September 11, 1980, without transfusion. patible blood group active glycolipids were absent from the uninvolved Blood Group and Pedigree. The patient's blood group was O, CCDee, mucosae. The results indicate that the cancer tissues possess the ability MN, Le(a-,b+). Her serum was reactive to all cells present in two to produce the globo series of glycolipids and the A active antigens but commercial panels. Since the reaction was strongly hemolytic, the that the surrounding uninvolved mucosae could not synthesize these presence of anti-PiPPk in a blood group p individual was suspected. glycolipids. The p blood group was confirmed serologically with anti-Pi, with ami Pk, and with sera from three individuals previously established as being of p phenotype by immunological and biochemical criteria (9). INTRODUCTION The patient denied consanguinous marriage of her parents. Her only sib had died and had had no offspring. She had had seven pregnancies The P blood group system is composed of five phenotypes, which terminated in spontaneous abortion between the third and fifth PI, P2, p, Pik, and P2k, and three different antigens, PI, P, and months of gestation. Pk, which are reported to be glycosphingolipids. PI, P, and Pk Pathologic Study. The surgical materials (Fig. 1) revealed two ulcer antigens were identified as galactosyl(al-4)lacto-JV-neotetrao- ous carcinomas, one (CI) on the lesser curvature of the upper cardia and another (CII) on the anterior wall of the antrum. The CI carcinoma sylceramide, globotetraosylceramide, and globotriaosylceram ide, respectively (1, 2). The p, Pik, and P2k phenotypes are was a moderately differentiated tubular adenocarcinoma with abundant fibrous stroma (Lauren's classification, intestinal type). CII was a extremely rare. The Pik phenotype lacks globotetraosylceram mucinous carcinoma with signet ring cells (Lauren's classification, ide, the P2k lacks both the PI antigen as well as globotetraosyl diffuse type). The two were completely separated from the healthy ceramide, and the p phenotype lacks globotriaosylceramide, gastric tissue. Both carcinomas had invaded the serosa, but lymph node globotetraosylceramide, and the I', antigen (3, 4). metastasis was not present in the surgical material. A report appeared in 1951 of a 66-year-old gastric cancer Isolation and Purification of Glycolipids. The procedures for the patient who had blood group p. Unexpectedly, the cancer tissue isolation and purification of the glycolipids were basically described in had the antigens which absorbed the antibody present in her a previous paper (7). Lipids were extracted from the tissue with 20 serum (5). The glycolipid fraction isolated from the cancer volumes each of chloroform :met hanol. 2:1, 1:1, and 1:9 (v/v), and the tissue was analyzed chemically and immunologically (6) and combined extracts were evaporated to dryness. The total lipids were was reactive with anti-Pi and anti-P|PPk antibodies. applied to a DEAE-Sephadex A-25 column (Pharmacia Fine Chemi cals, Uppsala, Sweden). The neutral lipids were eluted with chloro- We are interested in the glycolipids of human gastric cancer form:methanol:water (30:60:8, v/v) and the acidic lipids were eluted and described the presence of the incompatible A glycolipid with chloroform:methanol:0.8 M sodium acetate (30:60:8, v/v). The antigens in the blood group O cancer patients (7, 8). neutral glycolipids were isolated from the neutral lipid fraction accord We obtained surgical gastric specimens which consisted of ing to the method of Saito and Hakomori (10). The neutral glycolipids two different cancers and the surrounding uninvolved normal were further purified by silica gel column chromatography (Silica Gel tissue from a patient with blood group O, Le(a-,b+), and p. 60; Merck, Darmstadt, West Germany). The final purification of the individual neutral glycolipids was by preparative TLC2 (Silica Gel 60 This interesting case provided the opportunity to examine the glycolipid pattern of the stomach of an individual with the p plate; Merck) with chloroform:methanol:water, 65:25:4 or 60:35:8, as phenotype. It also provided an opportunity to determine the the solvent system. Analytical Procedures. TLC was carried out on Silica Gel 60 plates presence of incompatible blood group antigens in the cancer with chloroform:methanol:water (65:25:4, v/v; 50:40:8, v/v) as the Received 9/2/86; revised 11/24/86; accepted 12/17/86. solvent system. Carbohydrate composition and contents of the isolated The costs of publication of this article were defrayed in part by the payment glycolipid were determined by GLC of the trimethylsilyl ether deriva- of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. 2The abbreviations used are: TLC, thin layer chromatography; GLC, gas- 1To whom requests for reprints should be addressed. liquid chromatography. 1968 Downloaded from cancerres.aacrjournals.org on September 26, 2021. © 1987 American Association for Cancer Research. INCOMPATIBLE ANTIGENS IN p GASTRIC CANCER

DUODENUM ESOPHAGUS

Fig. 1. Schematic figure of surgical specimens of the stomach obtained from a patient of blood group O,Le(a-,b+),p. CI, moderately differentiated adenocarcinoma; ("II. mucinous carcinoma with signet ring cell; /, uninvolved mucosa; //, uninvolved mucosa. lives of the methylglycosid.es obtained after methanolysis at 95'C for 3 h with 3% IK'Irmethanol. GLC was performed with a glass column (3 mm inside diameter x 2 m) packed with 2% OV-1 programmed at 2'C/ min from 140 to 210Â°C. Immunochemical Analysis. Rabbit anti-Forssman antibody was ob tained as described previously (11). Human serum from a blood group _ A.p individual was donated by Dr. Ota (Department of Legal Medicine, Shinshu University School of Medicine). Rabbit anti-A and anti-B sera were purchased from Sankyo Junyaku Co. (Tokyo, Japan) and goat 12345 anti-Le* and anti-Leb sen; were from Ortho Diagnostics, Inc. (Raritan, NJ). Anti-H lectin (Ulex europeus) was purchased from E. Y. Labora Fig. 2. TLC of the neutral glycolipids of the gastric cancers and the uninvolved tories. Hemagglutination inhibition assays were performed using a 2% mucosae from a patient of blood group O,Le(a-,b+),p. Lane I, Forssman glyco suspension of RBC and approximately 5 ng of each glycolipid. Anti-A, lipid isolated from canine intestine; Lane 2, uninvolved mucosa (I); Lane 3, anti-B, anti-Le*, anti-Leb, and anti-Forssman sera were diluted to 4 uninvolved mucosa (II); Lane 4, cancer tissue (CI); Lane 5, cancer tissue (CII). units/0.025 ml and anti-H was diluted to 2 units/0.025 ml. Developing solvent chloroform:methanol:water, (65:25:4); visualization, cupric phosphoric acid reagent. TLC Immunostaining. The immunostaining procedure was a modifi cation of the method described by Uemura et al. (12). Approximately Table 1 Neutral glycolipids of the gastric cancers and the uninvolved mucosae of 50 ng of the total neutral glycolipid fraction were applied to a high a patient with blood group p performance aluminum sheet Silica Gel 60 TLC plate (Merck) and the The content of each neutral glycolipid separated by TLC was determined by plate was developed with a mixture of chloroform:methanol:water GLC using mannitol as an internal standard according to the method of Vance (60:35:8, v/v). Subsequently, the plate was blocked with 5% bovine and Sweeley (26). The values are expressed as Â¿imol/gprotein tissue. serum albumin in phosphate buffered saline and then incubated for 2 h Unin Gastric with the various antibodies described. The TLC plates was reincubated volved mu cancer Normal gastric for 2 h either with horseradish peroxidase-conjugated goat anti-rabbit cosa tissue mucosa [mean Â±SD IgG (The Jackson Laboratory, Bar Harbor, ME) or with peroxidase- I n a CII (n = 5)] conjugated goat anti-human IgG (Cappel) and washed with phosphate Ceramide monohexoside buffered saline. The chromatogram was incubated with a 4-chloro-l- Gal-Cer" 2.76 2.34 1.76 1.35 0.56 Â±0.15 naphthol:hydrogen peroxide solution (13). Glc-Cer Ceramide dihexoside Gal-Glc-Cer 1.84 1.65 1.25 0.92 0.24 Â±0.06 RESULTS Gal-Gal-Cer b 0.70 Â±0.07 Examination of the neutral glycolipids from the cancer tissues Ceramide trihexoside (CI and CII) and the uninvolved mucosae (I and II) by TLC is Gal-Gal-Glc-Cer 0.64 Â±0.09 shown in Fig. 2. Globotriaosylceramide and globotetraosylcer- Ceramide tetraglycoside amidc appeared to be absent and glucosyl or galactosylceramide GalNAc-Gal-Gal-Glc-Cer - 0.80 Â±0.24 Gal-GlcNAc-Gal-Glc-Cer 0.16 0.10 0.08 0.09 and lactosylceramide were present in both the cancer tissues 0.04 Â±0.02 and the uninvolved mucosae. The quantitative estimation of the Fucolipids 0.97 0.66 0.26 0.30 0.40 Â±0.12 glycolipid content is presented in Table 1. Differences were Total neutral glycolipids 5.73 4.75 3.35 2.66 3.37 Â±0.19 observed in both the tumor and the uninvolved mucosae from " Gal, galactose; C'er, ceramide; Glc, glucose; GalNAc, A'-acetylgalactosamine; the blood group p patient as compared to gastric mucosa from GlcNAc, iV-acetylglucosamine. normal individuals. Tissues from the blood group p patient did -, not detected. not have any detectable amount of globotriaosylceramide, globotetraosylceramide, and galabiosylceramide, which were the and the uninvolved mucosae. Le" and Leb glycolipids were main constituents of the total neutral glycolipids in the normal detected in both the cancer tissues and uninvolved mucosae. mucosae. The quantity of total neutral glycolipids in the unin To detect incompatible blood group antigens, we tested each volved mucosae was increased compared to the normal gastric fucolipid isolated from the cancer tissues and the uninvolved mucosae or the cancer tissues. mucosae for A and B blood group activities with a hemagglu- A TLC of the lacto series glycolipids isolated from the tination inhibition assay. As shown in Table 3, blood group A uninvolved mucosae (I) is shown in Fig. 3. The cancer tissues activities were found only in the CII cancer tissue and not in (CI, CII) and the uninvolved mucosae (I, II) have the same type the uninvolved mucosae (I and II) or the CI cancer tissue. No of fucolipids based upon characterization by GLC, TLC, and W-acetylgalactosamine was detected in fractions 5, 6, and 7 their blood group activities (Table 2). Blood group H active after a GLC analysis. Blood group B activity was not observed ceramide pentasaccharide was found in both the cancer tissues in any of the glycolipid fractions. 1969 Downloaded from cancerres.aacrjournals.org on September 26, 2021. © 1987 American Association for Cancer Research. INCOMPATIBLE ANTIGENS IN p GASTRIC CANCER

the uninvolved mucosae are shown in Fig. 5. Forssman glyco lipids were clearly detected only in the cancer tissues (CI and CII) but not in the uninvolved mucosae (I and II). These Forssman active glycolipids were migrating to the same position as the authentic Forssman glycolipid. Finally, to resolve the peculiar phenomenon of an individual reported by Levine et al. (5), the total neutral glycolipid frac tions of the cancer tissues and the uninvolved mucosae were analyzed with the human serum from a blood group A.p indi vidual which was expected to have anti-Pk (globotriaosylceramide), anti-P (globotetraosylceramide), anti-Pi antibody, and anti-Forssman antibody (Fig. 6). In the neutral glycolipid frac tions of the cancer tissues (CI, CII), regions of the plate corre sponding to globotriaosylceramide, globotetraosylceramide, and Forssman glycolipid standards were stained with the serum but there was no reaction with the uninvolved mucosae. There was no immunostaining materials migrating below the Forss man glycolipid with the serum of a blood group A,p individual (Fig. 6). The results suggested that the total neutral glycolipid fractions isolated from the cancer tissues contained only the Pk 12345678F (globotriaosylceramide) and P (globotetraosylceramide) anti gens but not the PI antigen. Fig. 3. TLC of the lacto scries glycolipids from the uninvolved mucosa (I). Lanes I and 2, lacto(neo)tetraosylceramide; Lanes 3 and 4, H active ceramide pentasaccharide: Lane 5, Le' active ceramide pentasaccharide; Lane 6, Le" active ceramide hexasaccharide; Lane 7, Leb active ceramide octasaccharide; Lane S. DISCUSSION more complex Ir1' active glycolipids: Lane F, Forssman glycolipid isolated from canine intestine (see Table 2). Developing solvent, chloroform:methanol:water The chemical and immunological characterization of the (55:40:8); visualization, cupric phosphoric acid reagent. glycolipids obtained from gastric cancer tissue of a patient with blood group p appears to be a rare event. In the present report, To confirm the presence of incompatible blood group activi the glycolipid composition of the gastric mucosa from a blood ties we utilized TLC-immunostaining using anti-A serum, rab group p individual was investigated and the presence of an bit anti-Forssman antibody, and human serum from a blood incompatible blood group antigen was observed in the cancer group p individual which contains unii-1',. Pk, and P antibodies. tissues. TLC-immunostaining with anti-A serum gave results similar to The uninvolved gastric mucosae lacked globotriaosylceram those for the hemagglutination inhibition assay. At least three ide (Pk antigen) and globotetraosylceramide (P antigen). These anti-A reactive glycolipids were observed with TLC-immuno glycolipids were also absent from erythrocytes (3), fibroblasts staining (Fig. 4). The upper spot corresponds to the Forssman (14) and a lymphoid cell line derived from a p phenotype glycolipid since preliminary' experiments showed that commer individual (15). Breimer et al. (16) showed that the stomach of cial rabbit anti-A serum slightly cross-reacted with Forssman a blood group p individual did not have any globotriaosylcer glycolipid on TLC immunostaining (data not shown). The amide or globotetraosylceramide by utilizing TLC analysis. middle spot had almost the same mobility as A" or Leb active The incompatible blood group antigens present in cancer ceramide hexasaccharide. The hemagglutination inhibition test tissues were observed and discussed by several workers (17, 18). seems to detect only this middle spot. The lower spot migrated The purified neutral glycolipids of the present case contained close to the Ab glycolipid. globotriaosylceramide and globotetraosylceramide while PI an The reactivities with the rabbit anti-Forssman antibody of tigen was not detected. Immunostaining revealed the presence the total neutral glycolipids fraction of the cancer tissues and of Pk and P antigens. The absence of the P! antigen should be

Table 2 Lacto series glycolipids of the gastric cancers and the uninvolved mucosae of a patient with blood group p The content of each glycolipid was determined by GLC using mannitol as an internal standard and was expressed as Â«imol/gprotein according to the method of Vance and Sweeley (26). Blood group activities of fucolipids were determined by hemagglutination inhibition assays as described in "Materials and Methods." Number on TLC indicates lacto series glycolipid fraction isolated from the cancer tissues and the uninvolved mucosae. protein)No.onTLC1,23,45678Major Tissue content (/Â¿mol/g mucosaI0.160.400.160.080.090.22II0.100.200.100.060.070.23CancertissueCI0.080.120.04tracetrace0.10CH0.090.140.07tracetrace0.09 structure)Gal-GlcNAc-Gal-Glc-Cer"Fuc-Gal-GlcNAc-Gal-Glc-CerGal-GlcNAc-Gal-Glc-Cer1FucFuc-Gal-GlcNAc-Gal-Glc-Cer1FucFuc-Gal-GlcNAc-Gal-GlcNAc-Gal-Glc-Cer1FucMoreglycolipid (tentative

complex Leb active glycolipidsBloodgroupHLe"LeÂ»LeÂ»Le"uninvolved ' Gai, galactose; GIcNAc, ,V-acetylglucosamine; Cer, ceramide: Fuc, fucose: Glc, glucose. 1970 Downloaded from cancerres.aacrjournals.org on September 26, 2021. © 1987 American Association for Cancer Research. INCOMPATIBLE ANTIGENS IN p GASTRIC CANCER

Table 3 Blood group A activities of Â¡actoseriesglycolipid fractions isolated from the gastric cancers and the uninvolved mucosae of a patient with blood group p Blood group A activity was determined by hemagglutination inhibition assay. - indicates that 200 Mg/ml of glycolipids could not inhibit the specific hemagglu tination. Numbers (pg/ml) in parentheses indicate the minimum amount of glycolipids completely inhibiting specific hemagglutination. For number on TLC, see Table 2.

No.JUINTLC cancerII Gastnc GbSCer- I12345678Â£? CII_ CI Gb4Cer- â€” â€”_ â€” ______+ (100)- - ++(50)+ (100)â€” â€” â€” 123456 12345 2 34 Fig. 6. TLC-immunostaining of the neutral glycolipids from the cancer tissues (CI and CII) and the uninvolved tissues (I and II) with the serum of a individual of blood group A.p. a, visualized with cupric phosphoric acid reagent. Lane I, standard glycolipids. Gb3Cer, globotriaosylceramide from human erythrocytes; a Gb4Cer, globotetraosylceramide from human erythrocytes. Lane 2, neutral gly colipids from A erythrocytes. Lane 3. cancer tissue (CI). Lane 4, cancer tissue (CII). Lane 5, uninvolved mucosa (I). Lane 6. uninvolved mucosa (II). b, stained with the serum of an individual of blood group A.p. Lane Â¡,standard glycolipids. Gb3Cer, globotriaosylceramide; Gb4Cer, globotetraosylceramide. Lane 2, cancer tissue (CI). Lane 3, cancer tissue (CII). Lane 4, uninvolved mucosa (I). Lane 5, uninvolved mucosa (II). c. Lanes 1 and 2, visualized with cupric phosphoric acid reagent. Lane I, standard glycolipids. GlcCer, glucosylceramide; LacCer, lacto- Gb4Cer,sylceramide; globotetraosylceramide; Gb3Cer, globotriaosylceramide; /â€¢'.Forssmanglycolipid. Gg3Cer, gangliotriaosylceramide; Lane 2, neutral glyco lipids from A erythrocytes. Lanes 3 and 4. stained with the serum of an individual of blood group A.p. Lanes 3 and 4 are the same glycolipids as Lanes I and 2, respectively.

Ab_ I. - confirmed by using a potent monospecific anti-Pi antibody. AS- Levine et al. (5) reported that the gastric cancer tissue from a Ad_ I patient with blood group p absorbed the patient's own antibod ies present in her serum and Kannagi et al. (6) purified the 123456 123456 glycolipid fraction from this cancer tissue. The upper phase Fig. 4. Immunostaining TLC of the neutral glycolipids from the cancer tissues obtained during the isolation of the neutral glycolipid fraction (CI and CII) and uninvolved mucosae (I and II) with rabbit anti-A serum, a, reacted with anti-P, and anti-P,PPk antibodies. They concluded visualized with cupric phosphoric acid reagent; h. stained with rabbit anti-A that the tumor had the capacity to synthesize 1', antigen but serum; Lane I. Forssman glycolipid isolated from sheep erythrocytes (F); Lane 2, neutral glycolipids of A erythrocytes (A', Ab, Ac, Ad); Lane 3, cancer tissue not the globo series glycolipids. Recently lizuka et al. (19) (CI); Lane 4, cancer tissue (CII); Lane 5, uninvolved mucosa (I); Lane 6. unin reported that UDP-Gal:LacCer al-4galactosyltransferase volved mucosa (II). which produces the Pk antigen was present in the lymphoblas- toid cells from a p blood type individual; however, Pk antigen cannot be produced in these cells. The present results indicate that in the cancer tissues of the p blood type individual this enzyme appears to be present and synthesizes the Pk antigen. GlcCer- Furthermore, Forssman antigenic activities were detected with TLC-immunostaining. The mobility of the Forssman active glycolipid on TLC was the same as that of the Forssman ceramide pentasaccharide of goat erythrocytes. The Forssman active glycolipids were associated only with the cancer tissues and were absent from the uninvolved normal mucosae. Hako- mori et al. (20) demonstrated that the Forssman activity was present in gastric cancer tissue from a Forssman negative population or uninvolved mucosa from a Forssman positive population although the uninvolved tissue from the Forssman negative population or the cancer tissue from the Forssman positive population did not show the activity. In this case, A activeglycolipidswere detected in the ceramide pentasaccharide to the ceramide octasaccharide fractions using 12345 12345 both TLC-immunostaining and hemagglutination inhibition Fig. 5. TLC-immunostaining of the neutral glycolipids of the cancer tissues (CI and CII) and the uninvolved mucosae (I and II) with rabbit anti-Forssman assay. Incompatible A antigens were observed histochemically antibody, a, visualized with cupric phosphoric acid reagent; b, stained with rabbit and chemically by several investigators (21-25). We also de anti-Forssman antibody. Lane I, standard glycolipids. GlcCer, glucosylceramide; scribed the blood group A active glycolipids of gastric cancer LacCer, lactosylceramide; GbiCer, globotriaosylceramide; Gb4Cer, globotetrao sylceramide; F, Forssman glycolipid. Lane 2, cancer tissue (CI). Lane 3, cancer tissue from a patient of blood type O (7, 8). Breimer (23) tissue (CII). Lane 4, uninvolved mucosa (I). Lane 5, uninvolved mucosa (II). reported that a gastric adenocarcinoma derived from a blood 1971 Downloaded from cancerres.aacrjournals.org on September 26, 2021. © 1987 American Association for Cancer Research. INCOMPATIBLE ANTIGENS IN p GASTRIC CANCER group B individual contained a difucosyl A structure as deter determination of Forssman and blood group A-active glycolipids in human gastric mucosa by inhibition assay of liposome lysis. J. Immunol. Methods, mined by direct probe mass spectrometry. However, the sample 53: 221-232, 1982. did not inhibit hemagglutination of A erythrocytes by anti-A 9. Ota, M., Kalo. M., Kanabako, F., and N'odii. K. A case of blood group p antisera. Clausen et al. (21) presented the new method to clarify phenotype (in Japanese). J. Jpn. Soc. Blood Transf., 27:451-453, 1981. 10. Saito, T., and Hakomori, S. Quantitative isolation of total glycosphingolipids these incompatible A antigens. Monofucosyl type 1 chain (type from animal cells. J. Lipid Res., 12: 257-259, 1971. 1 A") and difucosyl type 1 chain A (ALeb) have been described 11. Uemura, K., Yuzawa, M., and Taketomi, T. Immunochemical studies of lipids V. Effects of modified Hydrophobie moiety on immunogenicity and in colonie tumors of blood group O individuals by application immiinologÂ¡c;ilreactivity of Forssman glycolipid. Jpn. J. Exp. Med., 49: 1 - of carrier type specific monoclonal anti-A antibodies. The in 8, 1979. compatible A glycolipid in our study, detected by TLC-immu- 12. Uemura, K., Macher, B. A., DeGregorio, M., Scudder, P., Buehler, J., Knapp, Wâ€žand Feizi, T. Glycosphingolipid carriers of carbohydrate antigens of nostaining, is composed of at least two A active glycolipids. human myeloid cells recognized by monoclonal antibodies. Biochim. Bio phys. Acta, 846: 26-36, 1985. One has a lower K, than the Forssman glycolipid and seems to 13. Hawkes, R., Niday, ]â€¢'..,andGordon, J. A dot-immunobinding assay for be slightly different from the A' glycolipid on TLC migration. monoclonal and other antibodies. Anal. Biochem., 1/9: 142-147, 1982. The other had the mobility close to Ab glycolipid. This glyco 14. K-Ochiai, S., Naiki, M., and Makita, A. Defects of glycosyltransferase lipid was thought to be A/Leb, difucosyl type 1 chain A, as activities in human fibroblasts of Pk and p blood group phenotypes. Proc. Nati. Acad. Sci. USA, 74:5407-5410, 1977. concluded by Clausen et al. (21). 15. Momma, K., Shimoda, K., and Naiki, M. 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1972 Downloaded from cancerres.aacrjournals.org on September 26, 2021. © 1987 American Association for Cancer Research. Characterization of Glycolipids from the Gastric Cancer of a Patient of p,O,Le(1-,b+) Blood Type: Presence of Incompatible Blood Group Antigens in Tumor Tissues

Hiroshi Hattori, Kei-ichi Uemura, Hiroyuki Ogata, et al.

Cancer Res 1987;47:1968-1972.

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