Histochemical studies of human breast cancer using a monoclonal antibody against an oestrogen receptor-related antigen

Abstract

Summary The presence or absence of an oestrogen receptor-related antigen in breast tumours has been examined histochemically using a monoclonal antibody ('D59 -Coffer & King, 1981). In frozen sections, fixed either by the method of Tamura et al. (1980) or in methanol, staining was apparent in 14/24 (58%) and 22/26 (85%) of the breast cancers respectively. In paraffin sections fixed in ethanol, staining was present in 25/33 breast cancers (76%). In either type of section, staining was predominantly in the cytoplasm of the epithelial cells. When staining was scored by independent observers (2 or 3) and related to the tumour oestrogen receptor activity, determined by a standard biochemical technique, antigen was present in both receptorpositive and receptor-negative tumours. No significant association was found between the presence of antigen and receptors in the frozen sections, but for the series of paraffin sections, there was a weak association (r= +0.48) between the presence of the two proteins.Histochemical processing of paraffin sections from 9 tumours under conditions of higher sensitivity increased the staining significantly in 2/9 tumours, but did not alter the relationship between staining and receptor status.Six tissues were stained after exposure to 'receptor-translocating' conditions (250C/2 nM oestradiol/both for 1 h): this did not consistently change the subcellular staining pattern, though all tissues tended to stain more after exposure to 25°C.Staining was not blocked by absorption of the D, antiserum with a variety of pure proteins or human serum but at higher concentrations (approx. 2-15 mg protein ml -1), extracts from human uterus, an oestrogen-receptor-positive breast cancer and an oestrogen-receptor-negative breast cancer all effectively abolished staining in sections from another breast cancer.These results are consistent with other reports suggesting that the D5 antibody detects an antigen which is not the oestrogen receptor, but which may be associated with the receptor in its tissue distribution.In view of the value of oestrogen receptor measurements in the management of breast cancer (Hawkins, 1985), much effort has been expended in attempts to detect the receptor histochemically by a variety of methods (Lee, 1978;Pertschuk et al., 1979;Walker, Cove & Howell, 1980). For the purpose of immunohistochemistry, antibodies of both polyclonal (Tamura et al., 1984;Lope-Pihie et al., 1985) and monoclonal type (Greene et al., 1980;Coffer & King, 1981) have been generated and amongst these, one in particular (Greene et al., 1980) has been demonstrated to reflect accurately oestrogen receptor (R) status, in several different centres Pertschuk et al., 1985;Hawkins et al., 1986).In 1981, Coffer and King described two monoclonal antibodies (D, and C3) which they had raised against partially purified preparation of the oestrogen receptor protein from human myometrium. Of these antibodies, D5, in particular, has been the subject of further studies (Coffer et al., 1985a, b). In this paper, we report o...

Relationship between c-erbB-2 protein product expression and response to endocrine therapy in advanced breast cancer

“…PgR is one such protein, and indeed there is evidence that response rates are highest for tumours containing both ER and PgR (de Sombre et al, 1979;Hawkins, 1985; Osborne et al, 1980). However, studies correlating tumour levels of other oestrogen-regulated or -related proteins, such as ER-D5 related antigen and P24, with ER (Adams & McGuire, 1985;Cano et al, 1986;Giri et al, 1987;Hawkins et al, 1987;Home et al, 1988), prompting investigation of other possible predictors of response.…”

Immunoradiometric Studies with a Monoclonal Antibody against an Estrogen-Receptor-Related Protein in Human Breast Cancer

“…Recent work has reported the finding of good correlation between P29 and RE in human breast cancer samples, where P29 was detected by techniques requiring the use of a second antimouse peroxidase-conju gated antibody [3,4], In both tumor sections (IHC) and cytosol (ELISA), levels of P29 cor related with RE, but not RP. Clinical evi dence showing that P29 was at least as good as RE in the evaluation of patients' progno sis was also provided [3], In this paper we present data showing that this correlation is confirmed with an IRMA assay, as pre viously found in the preliminary study of Coffer et al [7], Compared with ELISA, advantages of this technique are the shorter time required for the assay and the direct quantification of the amount of D5 bound, thus avoiding inac curacy introduced by the use of a second antibody.…”

“…With these techniques we, and recently other investiga tors, have already reported the existence of a good correlation between P29 and RE, but not progesterone receptors (RP), in human breast cancer [3,4], We now present data in which P29 concentration is evaluated in hu man breast cancer cytosol through the use of an immunoradiometric assay (IRMA) with [l25I]-D5. The P29 values are compared with those of RE and RP, obtained by the classi cal binding assay with radiolabelled hor mone.…”