Increased detection of mammary carcinoma cells in marrow smears using antisera to epithelial membrane antigen

Abstract

Summary.-We have developed a technique for the immunocytochemical staining of marrow smears using antiserum to epithelial membrane antigen (EMA). This membrane component is confined to, but widely distributed in, epithelial tissues and tumours derived from them, and is strongly expressed by infiltrating breast carcinoma cells. Marrow aspirates from patients with both early and metastatic breast cancer have been examined, and the results of immunocytochemical staining compared with conventional cytology and histology. Staining with antiserum to EMA enabled us to detect small numbers of carcinoma cells, and increased the yield of positive samples. Furthermore, using this technique, we found malignant cells in the marrow of patients with primary breast cancer with no other evidence of metastatic disease. Thus immunocytochemical staining for EMA may be of value in the detection of micrometastases in patients with primary breast carcinoma.

Epithelial membrane antigen: Partial purification, assay and properties

“…An immunohistochemical stain for EMA can identify minute metastatic deposits of carcinoma in organs such as liver and bone marrow (Sloane et al, 1980;Dearnaley et al, 1981). Our most recent studies have shown that EMA positive cells can be found in the marrows of approximately one third of patients at the time of surgery for a primary breast cancer (Dearnaley et al, 1983).…”

A new monoclonal antibody to epithelial membrane antigen (EMA)-E29. A comparison of its immunocytochemical reactivity with polyclonal anti-EMA antibodies and with another monoclonal antibody, HMFG-2

“…Antisera to EMA have been used for the detection of bone marrow and lymph node metastases from breast carcinoma (Heyderman et al, 1979;Sloane et al, 1980a,b;Gugliotta et al, 1981;Dearnaley et al, 1981Dearnaley et al, , 1983Redding et al, 1983), and for identification of malignant cells in serous effusions Epenetos et al, 1982).…”

Evaluation of a rosetting method in detection of breast cancer cells

“…It has been shown that immunocytochemical staining for EMA can detect very small numbers of breast cancer in bone marrow smears that appear normal by conventional haematological techniques (Dearnaley et al, 1981).…”

“…Using this antibody to form rosettes 95-98% of marrow cells can be removed producing a smaller number of smears. We have compared the rosetting technique with density separation over lymphoprep previously described (Dearnaley et al, 1981 (Goding, 1976 (Beverley, 1980). Studies using cell-lines and human tissues had shown no crossreactivity with breast cancer or other epithelial tissues (Beverley, 1980 The marrow aspirate/RBC.SaM mixture, after removal of the Percoll cushion, was gently resuspended by inverting the tube 3 times.…”

“…Immunocytochemical staining for EMA has already been described (Dearnaley et al, 1981). Briefly, the endogenous alkaline phosphatase activity of osteoblasts was blocked by periodic acid, acetic acid and levamisole and the cells incubated with rabbit anti-EMA at 1:1000 dilution.…”