Immunocytochemistry and DNA-image cytometry in diagnostic effusion cytology I. Prevalence of markers in tumour cell positive and negative smears

Our plain-English summary of Immunocytochemistry and DNA-image cytometry in diagnostic effusion cytology I. Prevalence of markers in tumour cell positive and negative smears, by H. Motherby, M. Kube, N. Friedrichs, et al (Analytical Cellular Pathology, 1999).

The study

• Authors: H. Motherby, M. Kube, N. Friedrichs, B. Nadjari, K. Knops, A. Donner, B. Baschiera, P. Dalquen, A. Böcking
• Journal: Analytical Cellular Pathology
• Published: 1999;19(1):7-20
• Cited by: 16 articles, as counted by NIH iCite
• Read it: PubMed · Publisher · Free full text

Abstract

To determine the prevalence of immunocytochemical positivities for a panel of antibodies in benign and malignant cells in effusions with known follow-up in order to use these as diagnostic markers. Besides their ability to identify malignant epithelial cells their contribution to the differential diagnosis between carcinomatoses and mesotheliomas was investigated. 101 tumour cell positive and 53 negative effusions were stained with 12 different antibodies. Results were scored semiquantitatively per cell type. Furthermore, DNA-image cytometry was performed. While prevalence of Ber-EP4 positivity was 95.4% in metastatic carcinoma cells, it was 0% in those from mesotheliomas. No cell type reacted with this marker in benign effusions (0%). Ber-EP4 correctly differentiated between metastatic carcinoma and mesothelioma in 98.0%. Prevalence of DNA-aneuploidy was 95.4% in metastatic carcinomas, 57.1% in mesotheliomas and 0% in reactive effusions. Combining immunocytochemistry (Ber-EP4 positivity) and DNA-image cytometry (aneuploidy) results in a 100% detection of metastatic carcinomatoses and 57.1% of mesotheliomas. Both markers furthermore allowed a correct differentiation of these entities in 98%.

In plain English

This laboratory study from Düsseldorf, Germany, tested whether 12 antibody stains and DNA-image cytometry could detect cancer cells in effusions (body-cavity fluid) and tell metastatic carcinoma from malignant mesothelioma. Of 154 effusions collected April 1996 to December 1997, 101 were classified as containing tumour cells (87 carcinomas, 14 malignant mesotheliomas) and 53 as not, mostly by patient follow-up. Two positive cases without other evidence of cancer were classified by slide re-examination, confirmed by the tests under study. Ber-EP4 stained cells in 95.4% (83/87) of carcinoma effusions and in none of the 14 mesothelioma or 53 tumour-free effusions. Abnormal DNA content (aneuploidy) occurred in 83/87 carcinoma and 8/14 (57.1%) mesothelioma effusions, and in none of 52 tumour-free effusions in the DNA table. Both methods together detected 100% (87/87) of carcinoma and 57.1% of mesothelioma effusions. The authors proposed Ber-EP4 as a highly specific and sufficiently sensitive marker for epithelial cancer cells; they did not recommend an antibody panel for detecting malignant cells. To distinguish carcinoma from mesothelioma, they suggested adding anti-Leu-M1 and/or anti-CEA. They noted there is no agreed gold standard for checking such results.

We wrote this plain-English summary from the full article with the help of AI, and a second AI model checked each sentence against the article. It is not the authors' wording; the abstract and the article are their own account.

What the study did and found

• Design: Diagnostic laboratory study measuring how often immunocytochemical markers and DNA-aneuploidy occurred in effusion specimens. Results were checked against patient follow-up.
• Who was studied: 154 effusions (107 pleural, 43 peritoneal, three pericardial, one coul de sac aspiration), examined April 1996 to December 1997 at the Institute of Cytopathology, Düsseldorf. Patients came from the University Hospital of Düsseldorf and surrounding hospitals. By follow-up there were 87 carcinomatoses, 14 malignant mesotheliomas (without further histologic classification) and 53 effusions without tumour cells. One lymphoma case was excluded.
• Exposure: Not applicable. The study evaluated diagnostic tests on fluid samples: 12 antibodies (including Ber-EP4, Leu-M1, EMA and CEA) and DNA-image cytometry. Asbestos and other exposures were not studied.
• Main results:
  • Two cytologically positive cases had no further clinical, radiological or histopathological evidence of a malignant tumour. Slide re-evaluation by two expert cytopathologists led to a definitively positive diagnosis, which was confirmed by immunocytochemistry and DNA-cytometry.
  • Ber-EP4 staining was found in 95.4% (83/87) of carcinomatosis effusions, 0% (0/14) of mesothelioma effusions and 0% (0/53) of effusions without tumour cells.
  • Immunocytochemistry using Ber-EP4 and/or anti-Leu-M1 and/or anti-CEA correctly distinguished metastatic carcinomas from mesotheliomas in 98.0% (99/101) of effusions containing malignant cells.
  • EMA stained cells in 100% (14/14) of mesothelioma effusions, 97.7% (85/87) of carcinomatosis effusions and 28.8% (15/52) of effusions without tumour cells.
  • DNA-aneuploidy was found in 83/87 (95.4%) carcinomatosis effusions and 8/14 (57.1%) mesothelioma effusions. In the DNA-ploidy table, none of the 52 effusions without tumour cells was aneuploid; a later combined table gives 0/53.
  • Combining Ber-EP4 with DNA-cytometry detected 100% (87/87) of carcinomatoses. For mesotheliomas, detection was 0% with immunocytochemistry alone and 57.1% with DNA-cytometry.
  • 64.3% (9/14) of mesotheliomas had their greatest DNA stemline in the range 1.80c to 2.20c, compared with 9.2% (8/87) of carcinomatoses.
• The authors' conclusion: The authors proposed that Ber-EP4 staining may be used as a highly specific and sufficiently sensitive marker for neoplastic epithelial cells in effusions. They did not recommend a panel of antibodies for detecting malignant cells. To distinguish carcinomatoses from mesotheliomas, they suggested adding anti-Leu-M1 and/or anti-CEA, and they discussed anti-calretinin as a useful additional marker. They believed DNA-image cytometry can also increase the diagnostic accuracy of effusion cytology.
• Limitations the authors state:
  • The authors acknowledged that their negative-control approach was not optimal; they now use isotypic Ig as negative controls.
  • There is no general agreement on the gold standard for judging the accuracy of immunocytochemical or DNA-cytometric diagnoses.
  • Technical difficulties made anti-B72.3 and anti-HBME-1 staining almost impossible to evaluate, which led to high interobserver variability.
  • The four Ber-EP4-negative carcinoma effusions were too few to detect any correlation with the type of primary tumour.

Topics

The National Library of Medicine indexes this study under subjects our site covers: Mesothelioma.

Medical terms in this study, explained in our Lexicon: Cytology, Differential Diagnosis, Mesothelioma, Ber-EP4, Carcinoma.

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