A Novel Monoclonal Antibody (7B10) with Differential Reactivity between Human Mammary Carcinoma and Normal Breast1

A monoclonal antibody (7B10) which displays differential reactivity with breast carcinomas compared to benign lesions or normal breast tissue was selected by fusion of spleen cells from BALB/c mice immunized with the T47D human mammary carcinoma cell line. The antigen, rec ognized by 7B10 on T47D cells, appeared to be both surface and cyto plasm localized, as demonstrated by indirect immunofluorescence, immunoperoxidase, and electron microscopy studies. This antibody (It;*., ) bound with four human breast cancer cell lines (T47D, MCF7, ZR-75-1, and HSL53) which express estrogen receptors. No binding was observed with cancer cell lines of other origin or with normal cells. In vivo, by immunoperoxidase staining of frozen sections of normal breast, the antigen recognized by 7B10 appeared to be located on epithe lial cell membranes, whereas in benign and malignant mammary disor ders, staining also involved the cytoplasm, as confirmed by electron microscopy on fresh cancer tissue. On formalin-fixed, paraffin-embedded sections, cytoplasmic staining was detected in breast cancer, but no immunostaining was observed with benign lesions or normal breast. In paraffin sections, most normal tissues investigated did not react with 7B10 antibody. However, ducts in the parotid gland, tubules in the kidney and some biliary ducts, and apocrine glands in the skin showed irregular, diffuse weak staining. 7B10 was unreactive with adenocarcinomas of origin other than breast, except for some cells in ovarian clear cell carcinoma. No reactivity was observed with squamous carcinomas, lymphomas, or melanomas. The antigen recognized by 7B10 appeared to be a M, 32,000 protein, as identified by immunoprecipitation from extracts of T47D after labeling with |35S|methionine. Since the antigen was present only on the membrane of differentiated normal mammary epithelial cells, and was also expressed in the cyto plasm of tumor cells, it may be of interest in immunologie-ai studies of mammary epithelial cell differentiation. Moreover, since in formalinfixed tissues ¡mmunostaining is virtually confined to mammary carcino mas, monoclonal antibody 7B10 may have diagnostic applications in breast cancer.