Abstract
Human monoclonal antibodies specific for tumour-associated Thomsen-Friedenreich (TF) [Gal(β1–3)GalNAc(α)-O-] and Tn [GalNAc(α)-O-] glycoproteins were prepared using peripheral blood lymphocytes from healthy blood donors. The B lymphocytes were either directly transformed with Epstein-Barr virus (EBV) or transformed after an in vitro stimulation period with synthetic glycoproteins. The EBV-transformed lymphocytes were subsequently fused with a mouse-human heteromyeloma to secure antibody production and stability. IgM antibodies exhibiting different patterns of specificity for synthetic TF and Tn antigens were obtained, including antibodies specific for the α and β forms of different Gal(β1–3)GalNAc-O- and GalNAc-O- conjugates and antibodies agglutinating neuraminidase-treated erythrocytes. Several of the human monoclonal antibodies showed an increased binding to cultured carcinoma cells as compared to melanoma cells. This straightforward approach for the production of human monoclonal antibodies demonstrates the possibility of investigating the reactivity pattern of tumour-binding antibodies from peripheral blood lymphocytes. The binding patterns of these monoclonal antibodies show that healthy donors carry different fine specificities against synthetic TF/Tn antigens and that these antibodies react with different tumour cells.
Key words: In vitro immunization, Human monoclonal antibody, Thomsen-Friedenrich antigen, Tn antigen, T antigen
References
- 1.Borrebaeck CAK, Danielsson L, Möller SA. Human monoclonal antibodies produced by primary in vitro immunization of peripheral blood lymphocytes. Proc Natl Acad Sci USA. 1988;85:3995. doi: 10.1073/pnas.85.11.3995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 2.Caroll WL, Thielemans K, Dilley J, Levy R. Mouse × human heterhybridomas as fusion partners with human B cell tumors. J Immunol Methods. 1986;89:61. doi: 10.1016/0022-1759(86)90032-3. [DOI] [PubMed] [Google Scholar]
- 3.Clausen H, Stroud M, Parker J, Springer G, Hakomori S-I. Monoclonal antibodies directed to the blood group A associated structure, galactosyl-A: specificity and relation to the Thomsen-Friedenreich antigen. Mol Immunol. 1988;25:199. doi: 10.1016/0161-5890(88)90068-5. [DOI] [PubMed] [Google Scholar]
- 4.Danielsson L, Möller SA, Borrebaeck CAK. Effect of cytokines on specific in vitro immunization of human peripheral B lymphocytes against T-cell dependent antigens. Immunology. 1987;61:51. [PMC free article] [PubMed] [Google Scholar]
- 5.Dausset J, Moullec J, Bernard J. Acquired hemolytic anemia with polyagglutinability of red blood cells due to a new factor present in normal serum (anti-Tn) Blood. 1959;14:1079. [PubMed] [Google Scholar]
- 6.Hull SR, Carraway KL. Mechanism of expression of Thomsen-Friedenreicch (T) antigen at the cell surface of a mammary adenocarcinoma. FASEB J. 1988;2:2380. doi: 10.1096/fasebj.2.8.3360239. [DOI] [PubMed] [Google Scholar]
- 7.Itzkowitz SH, Yuan M, Montgomery CK, Kjeldsen T, Takahashi HK, Bigbee WL, Kim YS. Expression of Tn, sialosyl-Tn, and T antigens in human colon cancer. Cancer Res. 1989;49:197. [PubMed] [Google Scholar]
- 8.Lange PH, Winfield HN. Biological markers in urological cancer. Cancer. 1987;60:464. doi: 10.1002/1097-0142(19870801)60:3+<464::aid-cncr2820601506>3.0.co;2-c. [DOI] [PubMed] [Google Scholar]
- 9.Longenecker BM, Willans DJ, Maclean GD, Selvaraj S, Suresh MR, Noujaim AA. Monoclonal antibodies and synthetic tumor associated glycoconjugates in the study of the expression of Thomsen-Friedenriech like and Tn-like antigens on human cancers. J Natl Cancer Inst. 1987;78:489. [PubMed] [Google Scholar]
- 10.Ohlin M, Broliden PA, Danielsson L, Wahren B, Rosen J, Jondal M, Borrebaeck CAK. Human monoclonal antibodies against a recombinant HIV envelope antigen produced by primary in vitro immunization. Characterization and epitope mapping. Immunology. 1989;68:325. [PMC free article] [PubMed] [Google Scholar]
- 11.Ohlin M, Danielsson L, Carlsson R, Borrebaeck CAK. The effect of leucyl-leucine methyl ester on proliferation and Ig secretion of EBV-transformed human B lymphocytes. Immunology. 1989;66:485. [PMC free article] [PubMed] [Google Scholar]
- 12.Springer GF. T and Tn, general carcinoma autoantigens. Science. 1984;224:1198. doi: 10.1126/science.6729450. [DOI] [PubMed] [Google Scholar]
- 13.Springer GF, Desai PR, Scanlon EF. Blood group MN precursors as human brest carcinoma-associated antigens and “naturally” occurring human cytotoxins against them. Cancer. 1976;37:169. doi: 10.1002/1097-0142(197601)37:1<169::aid-cncr2820370124>3.0.co;2-#. [DOI] [PubMed] [Google Scholar]
- 14.Springer GF, Desai PR, Murthy MS, Tegtmeyer H, Scanlon EF. Human carcinoma associated precursor antigens of the blood group MN system and the host immune responses to them. Prog Allergy. 1979;26:42. [PubMed] [Google Scholar]
- 15.Stein R, Chen S, Grossman W, Goldenberg DM. Human lung carcinoma monoclonal antibody specific for the Thomsen-Friedenreich antigen. cancer Res. 1989;49:32. [PubMed] [Google Scholar]
- 16.Steuden I, Duk M, Czerwinski M, Radzikowski C, Lisowska E. The monoclonalantibody, antiasialoglycophorin from human erythrocytes, specific for beta-d-Gal-1-3-alfa-d-GalNAc chains (Thomsen-Friedenreich receptors) Glycoconjugate. 1985;2:303. [Google Scholar]
- 17.Wolf MF, Koerner U, Klump B, Schumacher K. Characterization of Thomsen-Friedenreich antibody subpopulations from normal human serum. Tumor Biol. 1987;8:264. doi: 10.1159/000217530. [DOI] [PubMed] [Google Scholar]
- 18.Yamada T, Fukui I, Yokokawa M, Oshima H. Changing expression of ABH blood group and cryptic T antigens of noninvasive and superficially invasive papillary transitional cell carcinoma of the bladder from initial occurance to malignant progression. Cancer. 1988;61:721. doi: 10.1002/1097-0142(19880215)61:4<721::aid-cncr2820610415>3.0.co;2-5. [DOI] [PubMed] [Google Scholar]