Skip to main content
NCBI home page
Japanese Journal of Cancer Research : Gann logoLink to Japanese Journal of Cancer Research : Gann
. 1988 Dec;79(12):1340–1348. doi: 10.1111/j.1349-7006.1988.tb01565.x

Establishment of a Human Monoclonal Antibody to Hanganutziu‐Deicher Antigen as a Tumor‐associated Carbohydrate Antigen

Osamu Usuba 1,2, Yoichi Fujii 3, Ichiro Miyoshi 4, Masaharu Naiki 5, Fujiro Sendo 1,✉
PMCID: PMC5917656  PMID: 3148605

Abstract

We have established a human‐human hybridoma producing a monoclonal antibody against a tumor‐associated carbohydrate‐specific antigen, Hanganutziu‐Deicher (HD) antigen. Human spleen lymphocytes from a patient with esophageal varices complicated with liver cirrhosis were cultured in serum‐free medium and co‐stimulated with both anti‐human μ‐chain antibodies and supernatants of concanavalin A‐stimulated human spleen cell culture (ConA sup). The activated lymphocytes were subsequently primed in vitro with particulate HD3 antigen and fused with a parent hybrid myeloma cell line, KR‐12. A hybridoma, 1F43E31G7 produced anti‐HD human monoclonal antibody (IgM λ). This monoclonal antibody reacted strongly with N‐glycolyl‐neuraminyl α2‐3 lactosylceramide (HD3) and slightly with N‐glycolylneuraminyl α2‐3 lactoneotetraosylceramide (HD3), but did not react with N‐glycolylneuraminyl α2‐3 lactoneohexaosylceramide (HD7), N‐acetylneuraminyl α2‐3 lactosylceramide (GM3) and other derivatives of HD3 prepared by chemical modification of the sialic acid residue of HD3, which indicates that the monoclonal antibody is directed precisely toward the terminal sialic acid and whole structure of HD3.

Keywords: HD antigen, Monoclonal antibody, Human hybridoma, Tumor‐associated antigen

Full Text

The Full Text of this article is available as a PDF (471.1 KB).

References

  • 1. ) Hanganutziu , M.Hémagglutinines hétérogénétiques apres injection de serum de cheval . Soc. Roum. Biol. , 91 , 1457 – 1459 ( 1924. ). [Google Scholar]
  • 2. ) Deicher , H.Über die Erzeugung heterospezifisher Hamagglutinine durch Injecktion artiremden Serums . Z. Hyg. Infektionkr. , 106 , 561 – 579 ( 1926. ). [Google Scholar]
  • 3. ) Stuart , C. A. , Grifin , A M. , Wheeler , K. M. and Batty , S.A thermostable antigen in beef‐cells . Proc. Soc. Exp. Biol. , 34 , 212 – 215 ( 1936. ). [Google Scholar]
  • 4. ) Davidsohn , I. and Walker , P. H.The nature of heterophile antibodies in infectious mononucleosis . Am. J. Clin. Pathol. , 5 , 455 – 465 ( 1935. ). [Google Scholar]
  • 5. ) Kasukawa , R. , Kano , K. , Bloom , M. L. and Milgrom , F.Heterophile antibodies in pathologic human sera resembling antibodies stimulated by foreign species sera . Clin. Exp. Immunol. , 25 , 122 – 132 ( 1976. ). [PMC free article] [PubMed] [Google Scholar]
  • 6. ) Yamakawa , T. and Suzuki , S.The chemistry of the lipids of post‐hemolytic residue or stroma of erythrocytes. I. Concerning the ether‐insoluble lipids of lyophilized horse blood stroma . J. Biochem. , 38 , 199 – 212 ( 1951. ). [Google Scholar]
  • 7. ) Higashi , H. , Naiki , M. , Matsuo , S. and Okouchi , K.Antigen of “serum sickness” type of heterophile antibodies in human sera; identification as gangliosides with N‐glycolylneuraminic acid . Biochem. Biophys. Res. Commun. , 79 , 388 – 395 ( 1977. ). [DOI] [PubMed] [Google Scholar]
  • 8. ) Merrick , J. M. , Schifferle , R. , Zadarlik , K. , Kano , K. and Milgrom , F.Isolation and partial characterization of the heterophile antigen of infectious mononucleosis from bovine erythrocytes . J. Supramol. Struct. , 6 , 275 – 290 ( 1977. ). [DOI] [PubMed] [Google Scholar]
  • 9. ) Higashi , H. , Nishi , Y. , Fukui , Y. , Ikuta , K. , Ueda , S. , Kato , S. , Fujita , M. , Nakano , Y. , Taguchi , T. , Sakai , S. , Sako , M. and Naiki , M.Tumor‐associated expression of glycosphingolipid Hanganutziu‐Deicher antigen in human cancers . Gann , 75 , 1025 – 1029 ( 1984. ). [PubMed] [Google Scholar]
  • 10. ) Hirabayashi , Y. , Higashi , H. , Kato , S. , Taniguchi , M. and Matsumoto , M.Occurrence of tumor‐associated ganglioside antigens with Hanganutziu‐Deicher antigenic activity on human melanomas . Jpn. J. Cancer Res. (Gann) , 78 , 614 – 620 ( 1987. ). [PubMed] [Google Scholar]
  • 11. ) Ohashi , Y. , Sasabe , T. , Nishida , T. , Nishi , Y. and Higashi , H.Hanganutziu‐Deicher heterophile antigen in human retinoblastoma cells . Am. J. Ophthalmol. , 96 , 321 – 325 ( 1983. ). [DOI] [PubMed] [Google Scholar]
  • 12. ) Higashi , H. , Ikuta , K. , Ueda , S. , Kato , S. , Hirabayashi , Y. , Matsumoto , M. and Naiki , M.Characterization of N‐glycolylneuraminic acid‐containing glycosphingolipids from a Marek's disease lymphoma derived chicken cell line, MSB1, as tumor‐associated heterophile Hanganutziu‐Deicher antigens . J. Biochem. , 95 , 785 – 794 ( 1984. ). [DOI] [PubMed] [Google Scholar]
  • 13. ) Naiki , M. , Fong , J. , Ledeen , R. and Marcus , D. M.The structure of human erythrocyte blood group P1 glycosphingolipid . Biochemistry , 14 , 4831 – 4837 ( 1975. ). [DOI] [PubMed] [Google Scholar]
  • 14. ) Vell , R. W. , Corfield , A. P. , Sander , M. and Schauer , R.Neuraminic acid‐specific modification and tritium labelling of gangliosides . Biochim. Biophys. Acta , 486 , 145 – 160 ( 1977. ). [DOI] [PubMed] [Google Scholar]
  • 15. ) Fujii , Y. , Higashi , H. , Ikuta , K. , Kato , S. and Naiki , M.Specificities of human heterophilic Hanganutziu Deicher (H‐D) antibodies and avian antisera against H‐D antigen active glycosphingolipids . Mol. Immunol. , 19 , 87 – 94 ( 1982. ). [DOI] [PubMed] [Google Scholar]
  • 16. ) Handa , S. and Nakamura , K.Modification of sialic acid carboxyl group of ganglioside . J. Biochem. , 95 , 1323 – 1329 ( 1984. ). [DOI] [PubMed] [Google Scholar]
  • 17. ) Mukuria , J. C. , Naiki , M. and Kato , S.Microstructure of the sialic acid moiety of N‐glycolylneuraminyllactosylceramide and the elucidation of Hanganutziu and Deicher (HD) antigenicity . Immunol. Lett. , 12 , 165 – 169 ( 1986. ). [DOI] [PubMed] [Google Scholar]
  • 18. ) Falkoff , R. J. M. , Zhu , L. and Fauci , A.Separate signals for human B cell proliferation and differentiation in response to Staphyloccocus aureus: evidence for a two‐signal model of B cell activation . J. Immunol. , 129 , 97 – 102 ( 1982. ). [PubMed] [Google Scholar]
  • 19. ) Mishell , B. B. and Shiigi , S. M.Preparation of mouse cell suspenisions . In “ Selected Methods in Cellular Immunology ,” pp. 4 – 27 ( 1980. ). W. H. Freeman Co. , San Francisco . [Google Scholar]
  • 20. ) Fujii , Y. , Inoue , T. , Ito , M. , Kimura , S. , Fuyama , S. , Arai , S. , Naiki , M. and Sendo , F.Heterogeneous profiles of factor that renders neutrophils cytotoxic obtained from a concanavalin A‐stimulated spleen cell culture in partial purification process . J. Immunol. , 136 , 3693 – 3699 ( 1986. ). [PubMed] [Google Scholar]
  • 21. ) Hirabayashi , Y. , Suzuki , T. , Suzuki , Y. , Taki , T. , Matsumoto , M. , Higashi , H. and Kato , S.A new method for purification of anti‐glycosphingolipid antibody Avian antihematoside (NeuGc) antibody. . J. Biochem. , 94 , 327 – 330 ( 1983. ). [DOI] [PubMed] [Google Scholar]
  • 22. ) Glassy , M. C. , Handley , H. H. , Hagiwara , H. and Royston , I.UC729‐6, a human lymphoblastoid B‐cell line useful for generating antibody‐secreting human‐human hybridomas . Proc. Natl. Acad. Sci. USA , 80 , 6327 – 6331 ( 1983. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 23. ) Kozbor , D. , Tripputi , P. , Roder , J. C. and Croce , C. M.A human hybrid myeloma for production of human monoclonal antibodies . J. Immunol. , 133 , 3001 – 3005 ( 1984. ). [PubMed] [Google Scholar]
  • 24. ) Oi , V. T. and Herzenberg , J. A.Immunoglobulin‐producing hybrid cell lines . In “ Selected Methods in Cellular Immunology ,” ed. Mishell B. B. and Shiigi S. M. , pp. 351 – 372 ( 1980. ). W. H. Freeman Co. , San Francisco . [Google Scholar]
  • 25. ) Tsu , T. T. and Herzenberg , L. A.Solid‐phase radioimmune assays . In “ Selected Methods in Cellular Immunology ,” ed. Mishell B. B. and Shiigi S. M. , pp. 373 – 397 ( 1980. ). W. H. Freeman Co. , San Francisco . [Google Scholar]
  • 26. ) Orlans , E.Fowl antibody II. The composition of specific precipitates formed by antisera to serum albumin, haemoglobin and some properties of non‐precipitating antibody . Immunology , 5 , 306 – 321 ( 1960. ). [PMC free article] [PubMed] [Google Scholar]
  • 27. ) Olsson , L. and Kaplan , H. S.Human‐human hybridomas producing monoclonal antibodies of predefined antigenic specificity . Proc. Natl. Acad. Sci. USA , 77 , 5429 – 5431 ( 1980. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 28. ) Schlom , J. , Wunderlich , D. and Teramoto , Y. A.Generation of human monoclonal antibodies reactive with human mammary carcinoma cells . Proc. Natl. Acad. Sci. USA , 77 , 6841 – 6845 ( 1980. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 29. ) Irie , R. F. , Sze , L. L. and Saxton , R. E.Human antibody to OFA‐I, a tumor antigen, produced in vitro by Epstein‐Barr virus‐transformed human B‐lymphoid cell lines . Proc. Natl. Acad. Sci. USA , 79 , 5666 – 5670 ( 1982. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 30. ) Glassy , M. C. , Handley , H. H. , Royston , I. and Lowe , D. H.Human monoclonal antibodies to human cancers . In “ Monoclonal Antibodies and Cancer ,” ed. Boss B. D. , Langman R. E. , Trombridge I. S. and Dulbecco R. , pp. 163 – 170 ( 1983. ). Academic Press; , New York . [Google Scholar]
  • 31. ) Edwards , P. A. W. , Smith , C. M. , Neville , A. M. and O'Hare , M. J.A human‐human hybridoma system based on a fast‐growing mutant of the ARH‐77 plasma cell leukemia derived line . Eur. J. Immunol. , 12 , 641 – 648 ( 1982. ). [DOI] [PubMed] [Google Scholar]
  • 32. ) Andreasen , R. B. and Olsson , L.Antibody‐producing human‐human hybridomas III. Derivation and characterization of two antibodies with specificity for human myeloid cells . J. Immunol. , 137 , 1083 – 1090 ( 1986. ). [PubMed] [Google Scholar]
  • 33. ) Andersson , J. , Moeller , G. and Sjoberg , O. B.lymphocytes can be stimulated by concanavalin A in the presence of humoral factors released by T cells . Eur. J. Immunol. , 2 , 99 – 101 ( 1972. ). [DOI] [PubMed] [Google Scholar]
  • 34. ) Andersson , J. , Edelman , G. M. , Moeller , G. and Sjoberg , O.Activation of B lymphocytes by locally concentrated concanavalin A . Eur. J. Immunol. , 2 , 233 – 235 ( 1972. ). [DOI] [PubMed] [Google Scholar]
  • 35. ) Kehrl , J. H. , Muraguchi , A. , Butler , J. L. , Falkoff , R. J. M. and Fauci , A. S.Human B cell activation, proliferation and differentiation . Immunol. Rev. , 78 , 75 – 96 ( 1984. ). [DOI] [PubMed] [Google Scholar]
  • 36. ) Muraguchi , A. , Kasahara , T. , Oppenheim , J. J. and Fauci , A. S.B cell growth factor and T cell growth factor produced by mitogen‐stimulated normal human peripheral blood T lymphocytes are distinct molecules . J. Immunol. , 129 , 2486 – 2489 ( 1982. ). [PubMed] [Google Scholar]
  • 37. ) Michael , J. H. R. and Michael , H. J.T‐dependent activation of resting B cells mediated by concanavalin A . Eur. J. Immunol. , 14 , 280 – 283 ( 1984. ). [DOI] [PubMed] [Google Scholar]
  • 38. ) Hawrylowicz , C. M. and Klaus , G. G. B.Activation and proliferation signals in mouse B cells IV. Concanavalin A stimulates B cells to leave G0, but not to proliferate . Immunology , 53 , 703 – 711 ( 1984. ). [PMC free article] [PubMed] [Google Scholar]
  • 39. ) DeFranco , A. L. , Raveche , E. S. , Asofsky , R. and Paul , W. E.Frequency of B lymphocytes responsive to anti‐immunoglobulin . J. Exp. Med. , 155 , 1523 – 1536 ( 1982. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 40. ) Muraguchi , A. , Butler , J. L. , Kehrl , J. H. and Fauci , A. S.Differential sensitivity of human B cell subsets to activation signals delivered by anti‐μ antibody and proliferative signals delivered by a monoclonal B cell growth factor . J. Exp. Med. , 157 , 530 – 546 ( 1983. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 41. ) Mishell , R. I. and Dutton , R. W.Immunization of dissociated spleen cell cultures from normal mice . J. Exp. Med. , 126 , 423 – 442 ( 1967. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 42. ) Click , R. E. , Benck , L. and Alter , B. J.Immune responses in vitro: culture conditions for antibody synthesis . Cell Immunol. , 3 , 264 – 276 ( 1972. ). [DOI] [PubMed] [Google Scholar]
  • 43. ) Hengartner , H. , Luzzate , A. L. and Schreier , M.Fusion of in vitro immunized lymphoid cells with X63 Ag8 . Curr. Top. Microbiol. Immunol. , 81 , 92 – 99 ( 1978. ). [DOI] [PubMed] [Google Scholar]
  • 44. ) Luben , R. A. and Mohler , M. A.In vitro immunization as an adjunct to the production of hybridomas producing antibodies against the lymphokine osteoclast activity factor . Mol. Immunol. , 17 , 635 – 639 ( 1980. ). [DOI] [PubMed] [Google Scholar]

Articles from Japanese Journal of Cancer Research : Gann are provided here courtesy of Wiley

RESOURCES