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Use of the Lectin from Amaranthus caudatus as a Histochemical Probe of Proliferating Colonic Epithelial Cells¹

Department of Internal Medicine, Veterans Administration Medical Center, University of Michigan School of Medicine, Ann Arbor, Michigan 48105 [C. R. B.]; Department of Biological Chemistry, University of Michigan School of Medicine, Ann Arbor, Michigan 48109 [Y-F. C., S. J. R., I. J. G.]; Human Tissue Resource, Department of Pathology, University of Maryland School of Medicine, Baltimore, Maryland 21201 [J. H. R.]; Department of Internal Medicine, Wayne State University School of Medicine, Detroit, Michigan 48201 [G. D. L.]; and Department of Preventative Medicine, Creighton University School of Medicine, Omaha, Nebraska 68178 [H. T. L.]

A newly isolated lectin, Amaranthus caudatus agglutinin (also called amaranthin or ACA), which binds to the Thomsen-Friedenreich antigen (T-antigen) and its sialylated variants, was used as a histochemical probe for proliferating cells in sections of human colonic tissues. Binding inhibition studies revealed that ACA binds to different sites on histological sections when compared to peanut agglutinin, which also recognizes the T-antigen. ACA bound selectively to the cells at the base of the colonic crypt [46 ± 4% (SEM) of glands] which is the zone of proliferation in this tissue and preferentially labeled cytoplasmic and apical membrane glycoconjugates. Only 7 ± 2% of the upper portions of the colonic crypts were labeled (P < 0.001 compared to the base), and this was largely a result of extensive labeling in 2 of 23 samples studies. A marked increase in histochemical labeling by ACA was seen in adenomatous polyps and adenocarcinomas of the colon, in which 82 ± 7 and 97 ± 2% of the glandular units were labeled, respectively. Transitional mucosa and connective tissue adjacent to cancers were also labeled by ACA. Neuraminidase studies indicated that removal of sialic acid residues enhanced binding by peanut agglutinin, but not ACA, to glycoconjugates in cancer specimens. Specimens of colonic tissue from patients with familial adenomatous polyposis (FAP) were examined with ACA; 83 ± 7% of adenomatous glands and 60 ± 7% of glands in flat, normal-appearing tissue were labeled. Colonic tissues from persons at 50% risk for hereditary nonpolyposis colorectal cancer (HNPCC), FAP, and normal colons were studied and given "weighted average" labeling scores that ranged from 0–400 to accommodate variable intensity and distribution of labeling. Normal colons had a weighted average score of 65 ± 33; FAP tissues had a score of 224 ± 76 (P < 0.001 compared to normal colon) and HNPCC tissues had a score of 74 ± 70 (P < 0.05 compared to normal colon). A group of five HNPCC cases had scores of 203 ± 43 (P < 0.001 compared to normal colon). ACA labels glycoconjugates in the proliferative region of normal human colonic epithelium and neoplastic lesions of the colon. The results of FAP and HNPCC tissues suggest that it may be useful for identifying foci of abnormal proliferation in familial colorectal cancer syndromes.

¹ Supported in part by the Research Service of the Veterans Administration and PHS Grants CA39233 (C. R. B.), GM29470 (I. J. G.), CA 45831 (G. D. L.), CA 42705 (H. T. L.), and Contract CP-95624 (J. H. R.), NIH, Department of Health and Human Services. S. J. R. was a Predoctoral Fellow supported by Pharmacological Sciences Training Grant 2T32-GM27767-11, NIH, Department of Health and Human Services, during the course of the work.

² To whom requests for reprints should be addressed, at GI Section (111D), VA Medical Center, 2215 Fuller Road, Ann Arbor, MI 48105.

Received 5/30/90. Accepted 10/29/90.

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