Immunohistochemical analysis of antiserum from rhesus monkeys immunized with human colon carcinoma

In an attempt to generate antibodies which recognize novel tumor-associated antigens we have immunized Rhesus monkeys (Macaca mulatta) with human colon carcinoma cells prepared from freshly excised tumors. Immunohistochemical characterization of polyclonal antisera from one monkey (DF6) revealed preferential reactivity with primary and metastatic colon carcinoma tissue, and a general lack of recognition of nonneoplastic mucosa. Immunoreactivity was localized to the luminal contents of glandular structures and to the apical surfaces of cells lining these glands. Immunoreactivity was not observed with any normal tissue examined. Examination of neoplastic tissues revealed reactivity with two gastric carcinoma specimens (n = 2) and one breast carcinoma (n = 7). In reactive colon carcinoma tissues, the pattern of staining with DF6 was similar to that of several other antibodies including anti-carcinoembryonic antigen, B72.3, anti-Le(x) and anti-Le(y). However, the panel of tissues recognized by these antibodies and DF6 differed significantly, suggesting that the DF6-reactive epitopes are unique. Human colon carcinoma cell lines maintained in vitro also expressed antigens recognized by DF6 in a pattern similar to that of surgically excised tissue. This preliminary characterization of DF6 antiserum suggests that immunization of Rhesus monkeys is a potentially useful protocol for identifying antigens preferentially expressed by human colon carcinoma.