Summary
Antisera and a lectin were used to demonstrate T-antigens in colorectal carcinomas. Rabbits were immunized with red blood cells on which the T-antigen had been exposed. The obtained polyclonal antiserum and a crude lectin (PNA) prepared from peanuts were used in an immunoperoxidase technique. The crude lectin and a chromatographic pure lectin showed the same staining specificity. Different fixatives, length of fixation and buffer compositions were tested on paraplast and frozen sections. Four per cent formaldehyde preserved the antigens well in paraplast embedded tissues when fixation was shorter than 48 h. In the immunoperoxidase technique the chromatogen used was 3-amino-9-ethyl-carbazole whose staining intensity was sensitive to the H2O2 concentration. Agglutination of T-exposed red blood cells was used to assess the anti-T titre of various sera. Normal animal serum contained anti-T antibodies, and the possibility of false positive reactions and methods to avoid it in immunohistochemistry is discussed.
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References
Anstee DJ (1981) The blood group MNSs-active sialoglycoproteins. Semin Hematol 18:13–31
Boland CR, Montgomery CK, Kim YS (1982) Alterations in human colonic mucin occurring with cellular differentiation and malignant transformation. Proc Natl Acad Sci USA 79:2051–2055
Bray J, Lemieux RU, McPherson TA (1981) Use of a synthetic hapten in the demonstration of the Thomsen-Friedenreich (T) antigen on neuraminidasetreated human red blood cells and lymphocytes. J Immunol 126:1966–1969
Bray J, Maclean GD, Dusel FJ, McPherson TA (1982) Decreased levels of circulating lytic anti-T in the serum of patients with metastatic gastrointestinal cancer: a correlation with disease burden. Clin Exp Immunol 47:176–182
Coon JS, Weinstein RS, Summers JL (1982) Blood group precursor T-antigen expression in human urinary bladder carcinoma. Am J Clin Pathol 77:692–699
Curran RC, Gregory J (1977) The unmasking of antigens on paraffin sections of tissue by trypsin. Experientia 33:1400–1401
De Lellis RA, Sternberger LA, Mann RB, Banks PM, Nakane PK (1979) Immunoperoxidase technichs in diagnostic pathology. Am J Clin Pathol 71:483–488
Graham RC, Lundholm U, Karnovsky MJ (1965) Cytochemical demonstration of peroxidase activity with 3-amino-9-ethyl-carbazole. J Histochem Cytochem 13:150–152
Howard DR, Taylor CR (1979) A method for distinguishing benign from malignant breast lesions utilizing antibody present in normal human sera. Cancer 43:2279–2287
Kania J, Uhlenbruck G, Janssen E, Klein PJ, Vierbuchen M (1980) Isolation, characterization and implications of anti-TF (Thomsen-Friedenreich) agglutinins from different sources. Immunobiol 157:154–168
Kim YD (1980) Immunochemical characteristics of human anti-T antibodies. Vox Sang 39:162–168
Klein PJ, Newman RA, Müller P, Uhlenbruck G, Schaefer HE, Lennartz KJ, Fischer R (1978) Histochemical methods for the demonstration of Thomsen-Friedenreich antigen in cell suspensions and tissue sections. Klin Wochenschr 56:761–765
Kuhlmann WD, Peschke P, Wurster K (1983) Lectin-peroxidase conjugates in histopathology of gastro-intestinal mucosa. Virchows Arch [Pathol Anat] 398:319–328
Mach J-P, Singh K, Perrinjaquet J, Carrel S, Merenda C, MacDonald D, Holburn A (1975) Relationship between carcinoembryonic antigen and blood group antigens A, B, and Lewis. Ann NY Acad Sci 259:389–403
Mepham BL, Frater W, Mitchell BS (1979) The use of proteolytic enzymes to improve immunoglobulin staining by the PAP technique. Histochem J 11:345–357
Ratcliffe RM, Baker DA, Lemieux RU (1981) Synthesis of the T-antigenic determinant in a form useful for the preparation of an effective artificial antigen and the corresponding immunoabsorbent. Carbohydr Res 93:35–41
Springer GF, Desai PR, Banatwala I (1975) Blood group MN antigens and precursors in normal and malignant human breast glandular tissue. JNCI 54:335–339
Springer GF, Desai PR, Murthy MS, Tegtmeyer H, Scanlon EF (1979) Human carcinoma-associated precursor antigens of the blood group MN system and the host's immune response to them. In: Ishizaka K, Kallós P, Waksman BH, de Weck AL (eds) Progress in allergy, vol 26. S Karger, Basel, pp 42–96
Springer GF, Murthy MS, Desai PR, Fry WA, Tegtmeyer H, Scanlon EF (1982) Patient's immune response to breast and lung carcinoma-associated Thomsen-Friedenreich (T) specificity. Klin Wochenschr 60:121–131
Sternberger LA (1979) Immunocytochemistry, 2nd ed. John Wiley & Sons Inc, New York
Taylor CR (1978) Immunoperoxidase techniques. Arch Pathol Lab Med 102:113–121
Uhlenbruck G, Pardoe GI, Bird WG (1969) On the specificity of lectins with a broad agglutinating spectrum. Z Immun Forsch 138:423–433
Uhlenbruck G (1981) The Thomsen-Friedenreich (TF) receptor: an old history with new mystery. Immunol Commun 10:251–264
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Supported by the Danish Cancer Society
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Örntoft, T.F. A method for T-antigen demonstration by a polyclonal antibody and peanut lectin; elimination of cross-reaction with naturally occurring antibodies. Vichows Archiv A Pathol Anat 404, 159–166 (1984). https://doi.org/10.1007/BF00704060
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DOI: https://doi.org/10.1007/BF00704060