- Email: [email protected]
TUMOUR MARKERS IN SEROUS EFFUSIONS
338 first studied this disease we too considered it mainly an aesthetic but we are now certain that women with this degree of disturbance of the vaginal. flora are at high risk of genital infection, especially after surgery. We find that infection of an episiotomy wound after delivery develops in about one-third of womenwith Aminkolpitis, and since we find Aminkolpitis in 5-10% of all sexually active women the condition is by no means unimportant. There are other indications that "non-specific vaginosis" plays an important part in other genital infections.4More attention should be paid to this disease.
(HMFG-2/ca 1)*
IMMUNOCHEMICAL STAINING RESULTS
we
problem,
Women’s Clinic, Klinikum der Albert-Ludwigs-Universität, D-7800 Freiburg, West Germany
EIKO E. PETERSEN
Institute for General Hygiene and Bacteriology, Zentrum Hygiene der Universität Freiburg
KLAUS PELZ
TUMOUR MARKERS IN SEROUS EFFUSIONS
SiR,-Studies of
the value of monoclonal antibodies for the immunocytochemical identification of neoplastic cells in serous effusions have not been consistent, particularly for mesothelial cells, and it is not clear whether discrepancies are attributable to variations in technique, differences in interpretation or both. One potential cause for discrepancy is the use by different groupss of smears of washed cells wet fixed in ethanol (or or smears of air-dried, unwashed cells fixed in acetone.3,4 We have done a small two-centre trial comparing these two methods. Specimens were chosen either because they were good examples of benign effusions containing numerous mesothelial cells or because they contained indubitable malignant cells. They were 4 processed by both washing and fixation and by Routine specimens arriving in both laboratories were fixed by both methods and spare slides were prepared for postage to the other centre so that immunocytochemical staining could be done in both laboratories on slides from the same specimen. Ten malignant and eight benign effusions were studied. The benign effusions were one peritoneal effusion due to liver cirrhosis and seven pleural effusions due to congestive cardiac failure (three), pneumonia (one), cardiac surgery (one), and unknown causes (two). The antibodies were HMFG-25 and Ca 1.6 The St Bartholomew’s Hospital laboratory used immunoperoxidase staining only while the4 Oxford laboratory used an immunoalkaline phosphatase and, in nine cases, the immunoperoxidase method as well (in these nine cases the same result was obtained by the two labelling
ether-ethanol) ,2
air-drying.4
procedure
techniques). The results (see table) indicate that the method of fixation makes substantial difference to the labelling reactions. The few discrepancies shown are probably explicable by minor technical differences. All slides were reviewed by members of the two laboratories, and even very weak staining was recorded. In some slides there were unsatisfactory thick areas in which staining appeared to be non-specific, and we also reached agreement over this. In two cases scattered plasma cells were non-specifically stained.4 In one case immunoperoxidase-stained smears contained no
*In each pair of results, HMFG-2 is recorded before the stroke and Ca 1 after; thus, ±/means weakly positive with HMFG-2 and negative with Ca 1. NS = non-specific staining in thick areas. tbreast, ovary, gastrointestinal tract, or lung.
positive eosinophil granulocytes, presumably because their content of endogenous peroxidase had not been fully inhibited. We conclude that both methods of cell processing are suitable for immunocytochemical labelling of serous effusions, at least when HMFG-2 and Ca 1 antibodies are used. The immunoperoxidase and immunoalkaline phosphatase methods give similar results, although the red reaction product of the latter method is easier to see than is the product of the peroxidase reaction, and may therefore be preferable when looking for small numbers of neoplastic cells. The fact that occasional mesothelial cells in some samples give positive reactions with HMFG-2, and, in two cases, feebly with Ca 1, does not substantially detract from the value of these antibodies for diagnosis, since both carcinoma and mesothelioma cells gave much stronger reactions with either of these antibodies, and could be
recognised as being morphologically different from the benign cells accompanying them. The discordant results obtained with Ca 1 by Pallesen et al7 may be attributable to the practice of adding alcohol to serous fluid samples before the cells are separated. ’
We thank Dr J. Taylor-Papadimitriou for supplying HMFG-2, Wellcome Diagnostics for supplying Ca 1, and Dr D. Y. Mason, in whose laboratory the Oxford series was processed. Nuffield
Department of Pathology, University of Oxford Laboratory of Clinical Cytology, Churchill Hospital,
1.
2.
3.
4.
5.
28: 17-21. 6. Ashall F, Bramwell ME, Harris H. A new marker for human antigen and the Ca 1 antibody. Lancet 1982; ii: 1-6
cancer
cells. 1: The Ca
A. I. SPRIGGS
Oxford
Department of Pathology, St Bartholomew’s Hospital,
M. CURLING
London EC1
IMMUNOCYTOCHEMICAL MARKERS IN CERVICAL SCREENING
4. Eschenbach
DA, Gravett MG, Hoyme UB, Holmes KK. Possible complications related to nonspecific vaginosis. 5th international meeting of the International Society for STD Research (Aug 1-3, 1983, Seattle, Washington) Epenetos AA, Canti G, Taylor-Papadimitriou J, Curling M, Bodmer WF. Use of two epithelium specific monoclonal antibodies for diagnosis of malignancy in serous effusions. Lancet 1982; ii: 1004-06. To A, Dearnaley DP, Ormerod MG, Canti G, Coleman DV. Indirect immunoalkaline phosphatase staining of cytologic smears of serous effusions for tumour marker studies. Acta Cytol 1983; 27: 109-13. Woods JC, Spriggs AI, Harris H, McGee JO’D. A new marker for human cancer cells 3: Immunocytochemical detection of malignant cells in serous fluids with the Ca 1 antibody. Lancet 1982; ii: 512-15. Ghosh AK, Spriggs AI, Taylor-Papadimitriou J, Mason DY. Immunocytochemical staining of cells in pleural and peritoneal effusions with a panel of monoclonal antibodies. J Clin Pathol 1983; 36: 1154-64. Taylor-Papadimitriou J, Peterson JA, Arklie J, Burchell J, Ceriani RL, Bodmer WF. Monoclonal antibodies to epithelium-specific components of the human milk fat globule membrane: production and reaction with cells in culture. Int J Cancer 1981;
ANNA K. GHOSH
SIR,-In the hope that the use of immunocytochemical tumourassociated markers might improve the specificity and sensitivity of cervical smear screening we studied specimens collected from women attending colposcopy, gynaecology, and antenatal clinics. Monolayer smears were prepared and stained with the Oxford tumour marker (Cal antibody) (100 cases) or monoclonal antibodies HMFG-1 and HMFG-22(78 cases) using an indirect immunoG, Jepsen FL, Hastrup J, Ipsen A, Hvidberg N. Experience with the Oxford marker (Ca 1) in serous fluids. Lancet 1983; i: 1326. Husain OAN, Millet JA, Grainger JM. Use of poly-L-lysine coated slides in the preparation of cell samples for diagnostic cytology with special reference to urine samples. J Clin Pathol 1980; 33: 309-11. Taylor-Papadimitriou J, Peterson JA, Arklie J, Burchell J, Ceriani RL, Bodmer WF. Monoclonal antibodies to epithelium specific components of the human milk fat globule membrane: production and reaction with cells in culture. Int J Cancer 1981;
7. Pallesen
tumour
1.
2.
28: 17-21
(HMFG-2/ca 1)*
IMMUNOCHEMICAL STAINING RESULTS
we
problem,
Women’s Clinic, Klinikum der Albert-Ludwigs-Universität, D-7800 Freiburg, West Germany
EIKO E. PETERSEN
Institute for General Hygiene and Bacteriology, Zentrum Hygiene der Universität Freiburg
KLAUS PELZ
TUMOUR MARKERS IN SEROUS EFFUSIONS
SiR,-Studies of
the value of monoclonal antibodies for the immunocytochemical identification of neoplastic cells in serous effusions have not been consistent, particularly for mesothelial cells, and it is not clear whether discrepancies are attributable to variations in technique, differences in interpretation or both. One potential cause for discrepancy is the use by different groupss of smears of washed cells wet fixed in ethanol (or or smears of air-dried, unwashed cells fixed in acetone.3,4 We have done a small two-centre trial comparing these two methods. Specimens were chosen either because they were good examples of benign effusions containing numerous mesothelial cells or because they contained indubitable malignant cells. They were 4 processed by both washing and fixation and by Routine specimens arriving in both laboratories were fixed by both methods and spare slides were prepared for postage to the other centre so that immunocytochemical staining could be done in both laboratories on slides from the same specimen. Ten malignant and eight benign effusions were studied. The benign effusions were one peritoneal effusion due to liver cirrhosis and seven pleural effusions due to congestive cardiac failure (three), pneumonia (one), cardiac surgery (one), and unknown causes (two). The antibodies were HMFG-25 and Ca 1.6 The St Bartholomew’s Hospital laboratory used immunoperoxidase staining only while the4 Oxford laboratory used an immunoalkaline phosphatase and, in nine cases, the immunoperoxidase method as well (in these nine cases the same result was obtained by the two labelling
ether-ethanol) ,2
air-drying.4
procedure
techniques). The results (see table) indicate that the method of fixation makes substantial difference to the labelling reactions. The few discrepancies shown are probably explicable by minor technical differences. All slides were reviewed by members of the two laboratories, and even very weak staining was recorded. In some slides there were unsatisfactory thick areas in which staining appeared to be non-specific, and we also reached agreement over this. In two cases scattered plasma cells were non-specifically stained.4 In one case immunoperoxidase-stained smears contained no
*In each pair of results, HMFG-2 is recorded before the stroke and Ca 1 after; thus, ±/means weakly positive with HMFG-2 and negative with Ca 1. NS = non-specific staining in thick areas. tbreast, ovary, gastrointestinal tract, or lung.
positive eosinophil granulocytes, presumably because their content of endogenous peroxidase had not been fully inhibited. We conclude that both methods of cell processing are suitable for immunocytochemical labelling of serous effusions, at least when HMFG-2 and Ca 1 antibodies are used. The immunoperoxidase and immunoalkaline phosphatase methods give similar results, although the red reaction product of the latter method is easier to see than is the product of the peroxidase reaction, and may therefore be preferable when looking for small numbers of neoplastic cells. The fact that occasional mesothelial cells in some samples give positive reactions with HMFG-2, and, in two cases, feebly with Ca 1, does not substantially detract from the value of these antibodies for diagnosis, since both carcinoma and mesothelioma cells gave much stronger reactions with either of these antibodies, and could be
recognised as being morphologically different from the benign cells accompanying them. The discordant results obtained with Ca 1 by Pallesen et al7 may be attributable to the practice of adding alcohol to serous fluid samples before the cells are separated. ’
We thank Dr J. Taylor-Papadimitriou for supplying HMFG-2, Wellcome Diagnostics for supplying Ca 1, and Dr D. Y. Mason, in whose laboratory the Oxford series was processed. Nuffield
Department of Pathology, University of Oxford Laboratory of Clinical Cytology, Churchill Hospital,
1.
2.
3.
4.
5.
28: 17-21. 6. Ashall F, Bramwell ME, Harris H. A new marker for human antigen and the Ca 1 antibody. Lancet 1982; ii: 1-6
cancer
cells. 1: The Ca
A. I. SPRIGGS
Oxford
Department of Pathology, St Bartholomew’s Hospital,
M. CURLING
London EC1
IMMUNOCYTOCHEMICAL MARKERS IN CERVICAL SCREENING
4. Eschenbach
DA, Gravett MG, Hoyme UB, Holmes KK. Possible complications related to nonspecific vaginosis. 5th international meeting of the International Society for STD Research (Aug 1-3, 1983, Seattle, Washington) Epenetos AA, Canti G, Taylor-Papadimitriou J, Curling M, Bodmer WF. Use of two epithelium specific monoclonal antibodies for diagnosis of malignancy in serous effusions. Lancet 1982; ii: 1004-06. To A, Dearnaley DP, Ormerod MG, Canti G, Coleman DV. Indirect immunoalkaline phosphatase staining of cytologic smears of serous effusions for tumour marker studies. Acta Cytol 1983; 27: 109-13. Woods JC, Spriggs AI, Harris H, McGee JO’D. A new marker for human cancer cells 3: Immunocytochemical detection of malignant cells in serous fluids with the Ca 1 antibody. Lancet 1982; ii: 512-15. Ghosh AK, Spriggs AI, Taylor-Papadimitriou J, Mason DY. Immunocytochemical staining of cells in pleural and peritoneal effusions with a panel of monoclonal antibodies. J Clin Pathol 1983; 36: 1154-64. Taylor-Papadimitriou J, Peterson JA, Arklie J, Burchell J, Ceriani RL, Bodmer WF. Monoclonal antibodies to epithelium-specific components of the human milk fat globule membrane: production and reaction with cells in culture. Int J Cancer 1981;
ANNA K. GHOSH
SIR,-In the hope that the use of immunocytochemical tumourassociated markers might improve the specificity and sensitivity of cervical smear screening we studied specimens collected from women attending colposcopy, gynaecology, and antenatal clinics. Monolayer smears were prepared and stained with the Oxford tumour marker (Cal antibody) (100 cases) or monoclonal antibodies HMFG-1 and HMFG-22(78 cases) using an indirect immunoG, Jepsen FL, Hastrup J, Ipsen A, Hvidberg N. Experience with the Oxford marker (Ca 1) in serous fluids. Lancet 1983; i: 1326. Husain OAN, Millet JA, Grainger JM. Use of poly-L-lysine coated slides in the preparation of cell samples for diagnostic cytology with special reference to urine samples. J Clin Pathol 1980; 33: 309-11. Taylor-Papadimitriou J, Peterson JA, Arklie J, Burchell J, Ceriani RL, Bodmer WF. Monoclonal antibodies to epithelium specific components of the human milk fat globule membrane: production and reaction with cells in culture. Int J Cancer 1981;
7. Pallesen
tumour
1.
2.
28: 17-21