title: Concurrent immunohistochemical staining of tumor-infiltrating lymphocytes and carcinoma-associated T (Thomsen-Friedenreich)/Tn antigens in human breast carcinoma.
abstract: The composition of tumor-infiltrating lymphocytes (TIL) often reflects the host's immune response to the tumor. To study the relationship of TIL and carcinoma-associated T/Tn antigens in breast carcinoma, a straightforward concurrent immunoenzyme staining procedure was developed. Fresh tissue was directly fixed in a zinc-based fixative to preserve lymphocyte markers and then routinely embedded in paraffin. The TIL subtypes in the sections were identified in the first immunostaining cycle by reaction with a monoclonal antibody (MAb) to lymphocyte markers CD3, CD4, CD8, CD19, or CD56, followed by a modified avidin-biotin procedure and diaminobenzidine tetrahydrochloride-H2O2 for color development. This was followed by paraformaldehyde fixation to block antibody crossreactivity. The T and Tn antigens on carcinoma cells were then demonstrated in a second staining cycle by reaction with an MAb against T or Tn antigen, followed by an indirect immunoalkaline phosphatase procedure and corresponding substrate systems for color development. The distinguishable brown color for TIL and blue or red color for T or Tn antigen enabled us to identify the TIL subsets and to describe their relations with T/Tn antigen expression in situ. This approach may contribute to better understanding of the patients' immune defenses against their tumor and aid in prognostication.
authors.affiliation: Heather M. Bligh Cancer Research Laboratories, Chicago Medical School, IL 60064, USA.
authors.forename: B L
authors.initials: BL
authors.lastname: Wang
authors.forename: G F
authors.initials: GF
authors.lastname: Springer
authors.forename: M W
authors.initials: MW
authors.lastname: Kaufman
mesh_terms: D015703:Antigens, CD / Q000145:classification / Q000276:immunology; D015295:Antigens, Tumor-Associated, Carbohydrate / Q000276:immunology*; D015415:Biomarkers; D001943:Breast Neoplasms / Q000276:immunology* / Q000473:pathology; D002240:Carbohydrate Sequence; D005260:Female; D006801:Humans; D007124:Immunoenzyme Techniques; D016246:Lymphocytes, Tumor-Infiltrating / Q000276:immunology*; D008969:Molecular Sequence Data
chemical_list: D015703:Antigens, CD; D015295:Antigens, Tumor-Associated, Carbohydrate; D015415:Biomarkers; C040286:Tn antigen; C016467:Thomsen-Friedenreich antigen