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PubMed1991

[Human breast cancer embedded in paraffin, fixed in Bouin's fluid. Immunohistochemical analysis of estrogen and progesterone receptors].

A new immunohistochemical method using a monoclonal estrogen receptor (ER) antibody and a progesterone receptor (PR) antibody has been performed to determine hormonal receptors on Bouin's liquid fixed and paraffin embedded tissue. This technique has been evaluated by comparison with conventional immunohistochemistry on frozen sections for 100 surgical biopsy specimens of breast carcinoma Erica (C) Prica (C); in 77 of these cases it was compared with enzyme immunoassay analyses (ER-EIA, PR-EIA). After a pretreatment by proteinase K for ER, the two specific monoclonal antibodies ER or PR were incubated overnight and revealed with a streptavidin biotin complex. The specific staining for ER and PR were exclusively located in the nuclei of cancer cells in both paraffin for ER and PR were exclusively located in the nuclei of cancer cells in both paraffin and frozen sections. The results on paraffin-embedded material were compared successively with ERICA (C) and ER-EIA and with PRICA (C) and PR-EIA. In 100 breast infiltrating carcinomas studied simultaneously on both paraffin and frozen sections, the concordance was 82% for ER and 85% for PR. In 77 breast carcinoma studied simultaneously on paraffin sections and EIA, the concordance was 74% for ER and 79.2% for PR. These results suggest that in the absence of frozen material, an immunohistochemical method on paraffin-embedded material can be used with a lower sensibility than on frozen sections. However, the detection of both ER and PR allows the identification of the hormondependent cases.

Acetates↗
PubMed1995

[Possibilities and limits of paraffin-embedded cell markers in diagnosis of primary cutaneous histiocytosis].

To date, the rare primary histiocytoses of the skin are diagnosed definitively on the basis of the clinical symptoms, H&E-stained sections, and demonstration of CD1 positivity in frozen sections and of Birbeck granules on electron microscopy. The improvement and analysis of antibodies with the ability to react in paraffin tissue allow retrospective evaluation and classification of these disorders. The antibodies for S-100-protein, peanut agglutinin (PNA) and PCNA (proliferating cell nuclear antigen) have been advocated for differentiation of the specific cells of Langerhans cell histiocytosis (LCH) from other histiocytic cell systems. To date the non-Langerhans cell histiocytoses (non-LCH) have no common ultrastructural and immunohistochemical characteristics. The infiltrate is made up of multiple cell populations, which are of significance for the cellular pathobiology (subtypes of monocytes/macrophages and dendritic cells). The number and distribution of the different monocyte/macrophages and dendritic cells and their ability to react with immunohistochemical markers in paraffin tissue can be completely different in different clinical entities. The antibodies against factor XIIIa (shown on xanthoma disseminatum) and the monoclonal antibody Ki-M1P (shown on juvenile xanthogranuloma) seem to be valuable in discrimination between LCH and non-LCH. Both markers show a positive staining pattern with the characteristic large macrophages. In juvenile xanthogranuloma, the foam cells and giant cells express Ki-M1P, KP1 and anti-cathepsin B. Other monocyte/macrophage markers with the ability to react in paraffin tissue, such as Mac387, lysozyme, alpha 1-antitrypsin and Leu-M1 (Anti-CD 15), in contrast, did not show a typical staining pattern with the characteristic large macrophages dominating the histological picture.

Biomarkers, Tumor↗
PubMed1996

Intermediate- and low-molecular-weight keratin detection with the monoclonal antibody MNF116. An immunohistochemical study on 232 paraffin-embedded cutaneous lesions.

Immunohistochemical detection of certain low to intermediate molecular weight keratins often is impaired in routinely processed specimens due to masking of these antigens by formalin fixation. Despite standard enzymatic digestion, AE1:AE3 and CAM 5.2, two of the most currently utilized antikeratin antibody preparations, either stain weakly or fail to stain basal keratinocytes and tumors composed of basaloid keratinocytes in paraffin sections of formalin-fixed tissue. We present here our experience with the monoclonal antibody MNF116 which detects keratins 5, 6, 8, 17, and 19 (DAKO, Carpinteria, CA). We have studied 232 routinely-processed skin lesions with MNF116 and compared the staining with that of AE1:AE3 mixture or CAM 5.2. In normal skin, the staining achieved with MNF116 was particularly strong on the basal cells of the epidermis and adnexae. MNF116 was positive in all 154 epithelial tumors and negative in all but one (a leiomyosarcoma) of 78 mesenchymal and melanocytic tumors. AE1:AE3 mixture was positive in all but four poorly-differentiated squamous cell carcinomas and it was only weakly positive in most basal cell carcinomas. CAM 5.2 was positive in tumors of the sweat apparatus, Merkel cell carcinomas, metastatic carcinomas, and 5/15 basal cell carcinomas. We consider that, in routinely processed specimens, MNF116 is very useful and convenient for detection of cytokeratin expression in cutaneous lesions, and therefore helpful in the evaluation of tumors with small cells and other poorly differentiated neoplasms of the skin.

Antibodies, Monoclonal↗
PubMed1998

Detection of P-glycoprotein in frozen and paraffin-embedded gastric adenocarcinoma tissues using a panel of monoclonal antibodies.

AIMS: Most chemotherapeutic regimens used against gastric carcinoma include anthracyclines whose effectiveness can be impaired by the presence of P-glycoprotein. In order to obtain a reliable pattern of P-glycoprotein expression in these tumours an immunohistochemical study using a panel of anti-P-glycoprotein antibodies was performed in frozen and paraffinized tissues. METHODS AND RESULTS: Frozen and paraffinized samples from 25 gastric carcinomas were immunohistochemically analysed using a panel of four anti-P-glycoprotein monoclonal antibodies including C219, MRK16, JSB-1 and C494. Semiquantitative analysis indicated that moderate or high P-glycoprotein levels were detected in 40% to 76% of gastric adenocarcinomas, depending on the anti-P-glycoprotein antibody used. The antibody C494 was the most sensitive in detecting P-glycoprotein in both frozen and paraffinized gastric carcinoma samples. Moreover, C494 showed a pattern of staining exclusively associated with the plasma membrane, in contrast to the cytoplasmic with reinforcement of plasma membrane pattern displayed by the other three antibodies. Significant differences in P-glycoprotein levels were obtained when C494 and MRK16 were used in frozen tissues. Finally, detection of P-glycoprotein in frozen samples did not improve when compared to paraffinized ones. CONCLUSIONS: It appears that P-glycoprotein is frequently expressed in gastric adenocarcinomas, and the use of C494 complemented by JSB-1 is recommended for reliable detection of P-glycoprotein in this neoplasm.

ATP Binding Cassette Transporter, Subfamily B, Mem↗