John Konrath

Immunohistochemical assessment of estrogen and progesterone receptors in stored imprints and cryostat sections of breast carcinomas

A peroxidase-antiperoxidase technique was used to visualize estrogen and progesterone receptors in stored imprints and cryostat sections of breast carcinomas that were prepared at the time of biopsy or frozen section diagnosis. This was done to provide an alternate technique for the assessment of the receptor status of tumors that could not be adequately assayed with other biochemical or immunocytological methods. Fixation in Zambonis fixative followed by passage through cold methanol and acetone before storage at -80 C insured good preservation of the receptor proteins over extended periods of time (up to 56 weeks). Immunostaining of these stored preparations with monoclonal antibodies against estrogen receptor (H222) and progesterone receptor (JZB39 and KD68) showed a high degree of correspondence with immunocytochemical assays (ER-ICA and PR-ICA) and biochemical analysis. This technique is easy to perform and provides reliable information, even in tumors that are too small and/or ill defined to permit separate sampling for receptor assays.

Heterogeneity vs. Asynchrony of Receptor Expression in Breast Cancer as Determined by Monoclonal Antibodies

The availability of monoclonal antibodies directed against estrogen and progesterone receptor proteins have made immunohistochemical study of these proteins in human breast tissue possible. Semiquantitative immunohistochemical analyses for estrogen receptor (ER) using H222 Sp correlated with quantitative biochemical receptor analyses. Two different monoclonal antibodies against progesterone receptor (PgR), B39 and KD68 were compared. Both were found to correlate with biochemical quantitation. Heterogeneity within breast tumors was investigated by comparing receptor expression with tumor histology and studying the effect of administered steroid hormone on receptor content. Both estrogen and progesterone receptor content was found to correlate with histologic grade. Within a given specimen, receptor expression often varied between different tumor components. A shift toward more homogeneous expression of PgR was observed in some tumors with estrogen pretreatment.

Ten Year Survival Patterns in Primary Breast Cancers Compared to Hormone Receptor Antigen Detection by Monoclonal Antibodies

The prognostic significance of immunocytochemical methods for estrogen (ER) and progesterone (PgR) receptor analyses was studied in 257 patients with primary breast cancer followed up to ten years. H222 antibody was used for ER and JZB39 used for PgR. ER positive (>10 fmol/mg prot) tumors using ligand assays showed disease free interval (DFI) advantage for 3 years but none after three years. H222 positive tumors were associated with survival advantage throughout the ten year period (p=0.004). PgR (by ligand analysis) advantage was only seen in the first 3.5 years. JZB39 positive tumors showed significant survival advantage through the ten year period (p=0.04). Both ER and PgR correlated with nuclear grade and histologic grade. Within histologic graDe S ER positive DFI advantage was observed only among HG II lesions (p=0.04). While separation within lymph node groups was observed in the initial period in each category, only the 1–3 node positive subgroup had distinct advantage throughout the period of follow-up. The data indicate that semi-quantitative immunocytochemical ER and PgR analyses contributes to the separation of prognostic groups among specific subgroups of primary breast cancer.