Abstract
A monoclonal antibody recognizing the blood group H type 2 antigen has been obtained from a BALB/c mouse immunized with MCF-7 (human mammary carcinoma) cells. The specificity of this antibody (A46-B/B10, IgM, kappa) has been identified by haemagglutination tests, immunohistochemistry, binding inhibition studies, and absorption experiments performed with synthetic oligosaccharides. The antibody is virtually nonreactive with H type 1 antigen or with closely related type 2 structures (e.g., Y antigen). A46-B/B10 strongly agglutinates human erythrocytes according to the amount of H substance expressed and can, therefore, easily discriminate between blood groups A1 and A2 as well as A1B and A2B (A1 and A1B are not or only weakly agglutinated). In immunohistochemistry, this antibody seems to provide a highly specific reagent for a restricted number of carcinomas and epithelial lineages in tissue sections and in vitro.
Highlights
BALB/c mice were immunized with live cells of the human mammary carcinoma cell line MCF-7, derived from a patient of blood group 0 (Soule et al, 1973)
Further examination against a panel of human cell lines (Table I) revealed the expression of this antigen on 4 out of 5 mammary carcinoma cell lines and on a mammary epithelial cell line derived from normal breast but transformed in vitro, H184A1
A46-B/BIO stained all of six carcinoma cell lines, whereas those derived from mesenchymal tissues were completey negative
Summary
Methodical details were as described in Karsten et al (1983). Briefly, BALB/c mice were immunized with live cells of the human mammary carcinoma cell line MCF-7, derived from a patient of blood group 0 (Soule et al, 1973). Spleen cells from one of these mice were fused with X63-Ag8.653 mouse myeloma cells (Kearney et al, 1979), and supernatants from hybridomas growing in HAT medium were screened by means of indirect immunofluorescence tests performed with MCF-7 and A-204 (human rhabdomyosarcoma, Giard et al, 1973) cells. Hybridoma A46-B/B10, which produced a mab reacting with a membrane antigen present on MCF-7 but absent on A-204 cells, was cloned repeatedly by limiting dilution on feeder cells, expanded and stored in liquid nitrogen. Cells of this clone were cultured in RPMI 1640 medium containing 5 x 10-5M 2-mercaptoethanol and 10% foetal calf or horse serum without antibiotics. Received 6 August 1987; and in revised form, 22 March 1988
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
