Our objective is the identification, isolation and characterization of cross-related antigens shared by human milk fat globule membrane (MFGM) and breast tumor membranes. Two general areas of research are planned to accomplish this objective. The first will concentrate on the characterization of proteins/glycoproteins isolated from human MFGM, preparation of antisera to these, and clarification of recognized determinants. The second will focus on the characterization, especially with emphasis on antibody-recognized determinants, of cross-related antigens of MFGM and membrane-enriched fractions of breast tumors which are recognized by a panel of monoclonal antibodies. Major techniques of protein/glycoprotein isolation will include molecular sieving and affinity (lectin and antibody-mediated) chromatography and preparative gel electrophoresis. Characterizations will include molecular weights, quantitative amino acid and carbohydrate analyses, and partial sequencing. Identification of antigenic determinants will be performed by comparisons of antibody-binding to glycoprotein, deglycosylated glycoprotein (chemical and enzymatic carbohydrate cleavage) and glycopeptides by Western and dot blotting and ELISA. A relatively large number of mono- and polyclonal antisera have been prepared versus MFGM and membrane-enriched breast tumor fractions which detect antigens common to both. The following salient characteristics of these antigens are pertinent to our proposed studies: (1) in general, these antigens are not tissue-specific and are expressed by secretory cells of the breast as well as by some exocrine and ductal epithelial cells; (2) when determined, all antisera generated thus far detect carbohydrate determinants; (3) MGFM-related antigens are generally expressed on the luminal surface of normal cells but assume a cytoplasmic localization in less-well-differentiated tumor cells; and (4) MFGM-related antigens from breast tumor cells possess lower molecular weights than corresponding antigen(s) in MFGM. Various human trial studies have recently been reported concerning the diagnostic, prognostic and therapeutic efficacy of antisera to MFGM and related tumor antigens. It is likely that the full potential of these reagents will only be realized when their recognized determinants are identified.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA044628-03
Application #
3187311
Study Section
Experimental Immunology Study Section (EI)
Project Start
1986-08-01
Project End
1989-07-31
Budget Start
1988-08-15
Budget End
1989-07-31
Support Year
3
Fiscal Year
1988
Total Cost
Indirect Cost
Name
Center for Molecular Medicine/Immunology
Department
Type
DUNS #
City
Belleville
State
NJ
Country
United States
Zip Code
07109
Ura, Y; Dion, A S; Williams, C J et al. (1992) Quantitative dot blot analyses of blood-group-related antigens in paired normal and malignant human breast tissues. Int J Cancer 50:57-63
Dion, A S; Smorodinsky, N I; Williams, C J et al. (1991) Recognition of peptidyl epitopes by polymorphic epithelial mucin (PEM)-specific monoclonal antibodies. Hybridoma 10:595-610
Williams, C J; Wreschner, D H; Tanaka, A et al. (1990) Multiple protein forms of the human breast tumor-associated epithelial membrane antigen (EMA) are generated by alternative splicing and induced by hormonal stimulation. Biochem Biophys Res Commun 170:1331-8
Major, P P; Dion, A S; Williams, C J et al. (1990) Breast tumor radioimmunodetection with a 111In-labeled monoclonal antibody (MA5) against a mucin-like antigen. Cancer Res 50:927s-931s
Dion, A S; Williams, C J; Herlyn, M et al. (1990) Human milk fat globule membrane glycoproteins express blood group-related determinants primarily on mucin-like epithelial membrane antigens and gp70. Biochem Int 22:295-302
Williams, C J; Major, P P; Dion, A S (1990) Enhanced expression and secretion of an epithelial membrane antigen (MA5) in a human mucinous breast tumor line (BT549). Tumour Biol 11:145-57
Wreschner, D H; Hareuveni, M; Tsarfaty, I et al. (1990) Human epithelial tumor antigen cDNA sequences. Differential splicing may generate multiple protein forms. Eur J Biochem 189:463-73
Ishida, M; Major, P P; Ura, Y et al. (1989) Related glycoproteins from normal secretory and malignant breast cells. Purification and initial comparative characterizations. Tumour Biol 10:12-22