Abstract
Lymph‐node lymphocytes of a patient with stomach cancer were fused with the mouse‐human heterohybridoma, HM‐S. A clonc (2F9) was isolated that showed stable production of an IgM antibody reactive with NUGC‐4 stomach cancer cell line. This antibody reacted predominantly with a cell surface antigen on cell lines originating from gastro‐intestinal cancer and adenocarcinoma of lung, whereas it was not generally reactive with other types of cancers, or with normal kidney cells or fibroblasts. Biotin‐labeled 2F9 antibody clearly stained cell smears and the nude mouse tumor of NUGC‐4, hut it did not show a positive reaction with stomach cancer tissues obtained from more than 10 patients, indicating that the antigen detected is very weakly expressed on tumor cells or on a limited number of stomach cancers. The antigen shed from NUGC‐4 cell line was detected in the culture supernatant. 2F9 antibody precipitated a glycoprotein with a molecular weight of over 200 kilodaltons as well as a possible glycolipid, from NUGC‐4 cells labeled with [3H]glucosamine or [35S]‐H2SO4. Periodic acid treatment of the tissue section decreased reactivity with 2F9 antibody, but heat, neuraminidase or protease treatment did not. These results suggested that the epitope is present on a carbohydrate moiety not containing sialic acid, and that a part of the antigen molecule is sulfated.
Keywords: Human monoclonal antibody, Stomach cancer, Cell surface antigen, Carbohydrate antigen
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REFERENCES
- 1. ) Olsson , L.Human monoclonal antibodies in experimental cancer research . J. Natl. Cancer Inst. , 75 , 397 – 403 ( 1985. ). [PubMed] [Google Scholar]
- 2. ) James , K. and Bell , G. T.Human monoclonal antibody production, current status and future prospects . J. Immunol Methods , 100 , 5 – 40 ( 1987. ). [DOI] [PubMed] [Google Scholar]
- 3. ) Koplowski , H. , Herlyn , D. , Lubeck , M. , Defreitas , E. and Sears , H. F.Human anti‐idiotype antibodies in cancer patients: is the modulation of the immune response bene‐fical for the patient? Proc. Natl. Acad. Sci. USA , 81 , 216 – 219 ( 1984. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 4. ) Frodin , J‐E. , Harmenberg , U. , Biberfeld , P. , Christensson , B. , Lefvert , A‐K. , Ringer , A. , Shetye , J. , Wahren , B. and Mellstedt , H.Clinical effects of monoclonal antibodies (MAb 17‐1A) in patients with metastatic colorectal carcinomas . Hybridoma , 7 , 309 – 321 ( 1988. ). [DOI] [PubMed] [Google Scholar]
- 5. ) Irie , R. F. and Morton , D. L.Regression of cutaneous melanoma by intralesional injection with human monoclonal antibody to ganglioside GD2 . Proc, Natl. Acad. Sci. USA , 83 , 8694 – 8698 ( 1986. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 6. ) Yoshikawa , K. , Ueda , R. , Obata , Y. , Utsumi , K. R. , Notake , K. and Takahashi , T.Human monoclonal antibody reactive to stomach cancer produced by mouse‐human hybridoma technique . Jpn. J. Cancer Res. , 77 , 1122 – 1133 ( 1986. ). [PubMed] [Google Scholar]
- 7. ) Ichimori , Y. , Sasano , K. , Itoh , H. , Hitotsumachi , S. , Kimura , Y. , Kaneko , K. , Kida , M. and Tsukamoto , K.Establishment of hybridomas secreting human monoclonal antibodies against tetanus toxin and hepatitis B virus surface antigen . Biochem. Biophys. Res. Commun. , 129 , 26 – 33 ( 1985. ). [DOI] [PubMed] [Google Scholar]
- 8. ) Akiyama , S. , Amo , H. , Watanabe , T. , Matuyama , M. , Sakamoto , J. , Imaizumi , H. , Kondo , T. and Takagi , H.Characteristics of three human gastric cancer cell lines, NU‐GC‐2, NU‐GC‐3 and NU‐GC‐4 . Jpn. J. Surg. , 18 , 438 – 446 ( 1988. ). [DOI] [PubMed] [Google Scholar]
- 9. ) O'Shannessy , D. J. , Dobersen , M. J. and Quarles , R. H.A novel procedure for labeling immunoglobulins by conjugation to oligosaccharide moieties . Immunol. Lett. , 8 , 273 – 277 ( 1984. ). [DOI] [PubMed] [Google Scholar]
- 10. ) Fagraeus , A. , Espmark , J. Å. and Jonsson , J.Mixed haemadsorption: a mixed antiglobulin reaction applied to antigens on a glass surface . Immunology , 9 , 161 – 175 ( 1965. ). [PMC free article] [PubMed] [Google Scholar]
- 11. ) Namikawa , R. , Mizuno , T. , Matsuoka , H. , Fukami , H. , Ueda , R. , Itoh , G. , Matsuyama , M. and Takahashi , T.Ontogenic development of T‐ and B‐cells and non‐lymphoid cells in the white pulp of human spleen . Immunology , 57 , 61 – 69 ( 1986. ). [PMC free article] [PubMed] [Google Scholar]
- 12. ) Gastro , J. M. D. C. , Deschamps , F. , Benbachir , M. , Henrichsen , J. , Volle , P. J. and Guinet , R. M. F.Highly sensitive biotin‐avidin sandwich ELISA for the rapid detection of pneumococcal capsular polysaccharide antigens . J. Immunol. Methods , 104 , 265 – 270 ( 1987. ). [DOI] [PubMed] [Google Scholar]
- 13. ) Woodward , M. P. , Young , W. W. , Jr. and Bloodgood , R. A.Detection of monoclonal antibodies specific for carbohydrate epitopes using periodate oxidation . J. Immunol. Methods , 78 , 143 – 153 ( 1985. ). [DOI] [PubMed] [Google Scholar]
- 14. ) Zola , H. , Moore , H. A. , Hohmann , A. , Humter , I. K. , Nikoloutsopoulos , A. and Bradley , J.The antigen of mature human B cells detected by the monoclonal antibody FMC7: studies on the nature of the antigen and modulation of its expression . J. Immunol , 133 , 321 – 326 ( 1984. ). [PubMed] [Google Scholar]
- 15. ) Ueda , R. , Ogata , S. , Morrissey , D. M. , Finstad , C. L. , Szkudlarek , J. , Whitmore , W. F. , Oettgen , H. F. , Lloyd , K. O. and Old , L. J.Cell surface antigens of human renal cancer defined by mouse monoclonal antibodies: identification of tissue‐specific kidney glycoproteins . Proc. Natl. Acad. Sci. USA , 78 , 5122 – 5126 ( 1981. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 16. ) Laemmli , U. K.Cleavage of structural proteins during the assembly of the head of bacteriophage T4 . Nature , 227 , 680 – 685 ( 1970. ). [DOI] [PubMed] [Google Scholar]
- 17. ) Teng , N. N. H. , Lam , K. S. , Riesa , R. C. and Kaplan , H. S.Construction and testing of mouse‐human heterohybridomas for human monoclonal antibody production . Proc. Natl. Acad. Sci. USA , 80 , 7308 – 7312 ( 1983. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 18. ) Ostberg , L. and Pursch , D.Human × (mouse × human) hybridomas stably producing human antibodies . Hybridoma , 2 , 361 – 367 ( 1983. ). [DOI] [PubMed] [Google Scholar]
- 19. ) Fong , S. H. H. , Perkins , S. , Raubitschek , A. , Larrick , J. , Lizak , G. , Fishwild , D. , Engleman , E. G. and Grumet , F. C.Rescue of human monoclonal antibody production from an EBV‐transformed B cell line by fusion to human‐mouse hybridoma . J. Immunol. Methods , 70 , 83 – 90 ( 1984. ). [DOI] [PubMed] [Google Scholar]
- 20. ) Cahan , L. D. , Irie , R. F. , Singh , R. , Cassidenti , A. and Paulson , J. C.Identification of a human neuroectodermal tumor antigen (OFA‐I‐2) as ganglioside GD2 . Proc. Natl. Acad. Sci. USA , 79 , 7629 – 7633 ( 1982. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 21. ) Tai , T. , Paulson , J. C. , Cahan , L. D. and Irie , R. F.Ganglioside GM2 as a human tumor antigen (OFA‐I‐1) . Proc. Natl. Acad. Sci. USA , 80 , 5392 – 5396 ( 1983. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 22. ) Yamaguchi , H. , Furukawa , K. , Fortunato , S. R. , Livingston , P. O. , Lloyd , K. O. , Oettegen , H. F. and Old , L. J.Cell‐surface antigens of melanoma recognized by human monoclonal antibodies . Proc. Natl. Acad. Sci. USA , 84 , 2416 – 2420 ( 1987. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 23. ) Cote , R. J. , Morrissey , D. M. , Houghton , A. N. , Thomson , T. M. , Daly , M. E. , Oettegen , H. F. and Old , L. J.Specificity analysis of human monoclonal antibodies reactive with cell surface and intracellular antigens . Proc. Natl. Acad. Sci. USA , 83 , 2959 – 2963 ( 1986. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 24. ) Hirohashi , S. , Clausen , H. , Nudelman , E. , Inoue , H. , Shimosato , Y. and Hakomori , S.A human monoclonal antibody directed to blood group i antigen: heterohybridoma between human lymphocytes from regional lymph nodes of a lung cancer patiënt and mouse myeloma . J. Immunol. , 136 , 4163 – 4168 ( 1986. ). [PubMed] [Google Scholar]
- 25. ) Schrump , D. S. , Furukawa , K. , Yamaguchi , H. , Lloyd , K. O. and Old , L. J.Recognition of galactosylgloboside by monoclonal antibodies derived from patients with primary lung cancer . Proc. Natl. Acad. Sci. USA , 85 , 4441 – 4445 ( 1988. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 26. ) Kjeldsen , T. B. , Rasmussen , B. B. , Rose , C. and Zeuthen , J.Human‐human hybridomas and human monoclonal antibodies obtained by fusion of lymph node lymphocytes from breast cancer patients . Cancer Res. , 48 , 3208 – 3214 ( 1988. ). [PubMed] [Google Scholar]
- 27. ) Saito , H. , Uchiyama , K. , Nakamura , L , Hiraoka , H. , Yamaguchi , Y. and Taniguchi , M.Characterization of a human monoclonal antibody with broad reactivity to malignant tumor cells . J. Natl. Cancer Inst. , 80 , 728 – 734 ( 1988. ). [DOI] [PubMed] [Google Scholar]
- 28. ) Morrison , S. L.Transfectoma provide novel chimeric antibodies . Science , 229 , 1202 – 1207 ( 1985. ). [DOI] [PubMed] [Google Scholar]
- 29. ) Nishimura , Y. , Yokoyama , M. , Ueda , R. , Kudo , A. and Watanabe , T.Recombinant human‐mouse chimeric monoclonal antibody specific for common acute lymphocytic leukemia antigen . Cancer Res. , 47 , 999 – 1005 ( 1987. ). [PubMed] [Google Scholar]