Erythrocyte polyagglutination antigens T and Tn are truncated O-glycan chai
ns that are also carcinoma-associated antigens, We investigated whether Tk
polyagglutination antigen could similarly be a carcinoma-associated marker
and a target of immunotherapy. Monoclonal antibody LM389 was raised against
Tk erythrocytes and tested by immunohistochemistry, LM389 strongly reacted
with 48% human colorectal carcinomas. Labeling of normal tissues was visib
le on epithelial cells, mainly digestive, but was confined at a supranuclea
r level. Expression of the antigen on cloned human carcinoma cells correlat
ed with sialosyl-Tn expression. O-Sialoglycoprotein endopeptidase treatment
revealed that on carcinomas and cell lines, the epitope was present on O-g
lycans. Antibody specificity was determined using synthetic carbohydrates.
Direct binding and inhibition studies indicated that LM389 best ligands wer
e terminated by two branched N-acetylglucosamine units. Screening of murine
cellular cell lines with LM389 allowed development of an experimental mode
l with Tk-positive and -negative cells in syngeneic BDIX rats. Vaccination
of rats with Tk erythrocytes provided a protection against growth of rat Tk
-positive, but not of Tk-negative, tumor cells in association with the deve
lopment of antibodies. Taken together, the results indicate that Tk polyagg
lutination antigen is a new colorectal carcinoma-associated antigen, absent
from the normal cell surface, resulting from alteration of O-glycans biosy
nthesis and with potential as a target of immunotherapy.