Br. J. Cancer
X, 67 69 (I990), Br. 62, J. Suppl. Cancer (1990),
62,
Suppl.
X,
67
69
1990 Ltd., 1990~~~~~~~~
11" Macmillan Press © Macmillan Press Ltd.,
Immunohistochemical identification of CA125 and the F(ab'), fragments of the murine monoclonal antibody OC125 in ovarian cancer tissue G. Ronay', W. Jigerl, K. Weiss', H. Feistel2, F. Wolf2, A.H. Tulusan' & N. Lang' 'University of Erlangen, Department of Obstetrics and Gynecology, Universitdtsstrasse 21-23 and 2Department of Nuclear Medicine, Krankenhausstrasse 12, 8520 Erlangen, FRG. Summary Benign and malignant tissue samples of 19 ovarian cancer patients who received an intravenous infusion of radiolabelled F(ab')2 fragments of OC125 were examined for expression of CA125 and the cellular location of the injected antibodies. CA125 could be detected by immunohistochemistry in 47 of 71 biopsies containing cancer cells, but not in 258 biopsies with benign tissue. F(ab')2 fragments were found in 34 of these 47 biopsies. The diffuse intracytoplasmic staining pattern seemed to indicate that the injected antibodies had crossed the membranes of the cancer cells. Since the antibody could not be detected in benign tissues, further applications of OC125 for diagnostic and therapeutic purposes seems to be justified.
In 1981 Bast et al. described the murine monoclonal antibody (MAb) OC125 which bound to an antigen expressed on an ovarian cancer cell line. The antigen, which was coded CA125, was also detected in tissue sections of human epithelial ovarian cancers but not in the normal ovary (Kabawat et al., 1983; Dietel et al., 1986). It was therefore postulated that this specificity of the expression of CA125 could be used for the detection of ovarian cancerous tissues in patients. Radiolabelled fragments of OC125 were used for immunoscintigraphy and areas of increased radioactivity during scintigraphy were interpreted as specific accumulation of the antibodies to CA125 producing tissues, e.g. ovarian cancer. Thus preoperative information on tumour localization and metastatic spread could be achieved (Goldenberg et al., 1978, 1980; Epenetos et al., 1982; Pimm et al., 1985; Chatal et al., 1987; Gitsch & Pateisky, 1987). Recently intraoperative tracing has also been applied successfully (Jager et al., 1987, 1988). The distribution of the infused antibodies to different tissues has, however, so far only been described in terms of comparing the radioactivity localized in benign and malignant tissues (Haisma et al., 1988). In order to evaluate the binding of the injected OC125 antibody fragments we performed immunohistochemical staining procedures in different tissues obtained during surgery several days after immunoscintigraphy.
Patients, materials and methods
Antibody infusion In 19 ovarian cancer patients 1 mg of "3'I-labelled F(ab')2 fragments of the murine monoclonal antibody OC125 were infused at the Department of Nuclear Medicine, University of Erlangen as described previously (Feistel et al., 1988; Jiger et al., 1988). In two of these patients the antibodies were infused before primary surgery to a suspected ovarian cancer in order to evaluate if lymph node metastases could be detected. In the remaining 17 patients antibodies were infused prior to second-look operations to localize remaining tumour after chemotherapy or recurrence of the disease (Jager et al., 1988). Surgery Surgery was performed 7-13 days after infusion of the antibodies. During surgery biopsies were taken from different parts of the abdomen. Radioactivity was measured during surgery by a hand-held probe and biopsies which were Correspondence: G. Ronay. Supported by Istopen Diagnostik GmbH, Dreieich, FRG.
removed because of higher radioactivity compared with background were sampled (Jaiger et al., 1987). Further biopsies were resected from areas with background radioactivity as controls. All biopsies were fixed in Carnoy's solution (six parts ethanol 96%, three parts chloroform and one part glacial acetic acid) (Taylor, 1986) for 16-24 h according to the size of the tissue sample. Thereafter tissues were embedded in paraffin (Paraplast) at 56'C and stored until histological examination. Immunohistochemical staining of CA125 From each biopsy obtained, three tissue sections were prepared with a depth of 5 tm. One of the sections was used for routine haematoxylin staining procedures, while the remaining two sections were used for immunohistochemical staining according to the avidin-biotin-immunoperoxidase (ABC) technique (Hsu et al., 1981). Briefly, the samples were rehydrated for 3 x 10 min in xylol and 4 x 1 min in alcohol. Thereafter endogenous peroxidase was blocked by the addition of 0.3% H202-methanol solution for 20min. Since our previous studies seemed to demonstrate a partial destruction of the CA125 antigen by proteinases, incubation of tissues with trypsin and pronase was omitted during the study. To avoid non-specific binding of MAb OC125 via Fc receptors, the slides were treated before incubation with MAb with non-immunized sheep serum. After rinsing the tissue sections for 5 min in phosphate buffered saline (PBS), one of the two parallel sections was incubated with the undiluted murine MAb OC125 overnight at 4'C. During this time the parallel section remained stored until further processing in PBS. After this procedure a further incubation of both sections with the biotinylated rabbit antimouse antibody (dilution 1:50) and the streptavidine-biotin-peroxidase complex (dilution 1:100) (Amersham, FRG) was performed. The incubation period in each case was 30 min. The sections were rinsed for 3 x 5 min with PBS after every individual incubation step. Further reaction with 0.5% 3,3-diaminobenzidinetetrahydrochloride (DAB)/0.02% H202 solution resulted in the formation of a brown precipitate. Nuclear contrast staining was carried out using Mayer's haematoxylin. Immunohistochemical staining of the 0C125 F(ab')2 fragments For these experiments slices of human amniotic epithelium were used. The expression of CA125 in amniotic tissue was examined by immunohistochemistry using the monoclonal antibody OC125 as described above. To evaluate if the biotinylated rabbit antimouse antibodies also bound to the F(ab')2 fragments of OC125, amniotic epithelium was incubated with the OC125 F(ab')2 fragments
68
G. RONAY et al.
for 24 h. Thereafter the rabbit antimouse antibody was added to the tissue section and further staining was performed with the ABC detection system in the usual way. The OC125 F(ab')2 fragments were obtained from serum of patients, sampled 10 min after infusion of the antibodies. All experiments were repeated after storage of the serum for 14 days at 4°C. To exclude non-specific staining, amniotic epithelium was incubated with the ABC detection system without application of the OC125 or its fragments.
Results With the usual staining procedure the presence of the CA125 antigen in amniotic epithelium could be demonstrated. Using the OC125 fragments identical staining was observed, even when the stored sera were used (Figure 1). In the control experiment without the OC 125 antibodies no staining was detected. Three hundred and twenty-nine biopsies were examined for CA125 expression and presence of the F(ab')2 fragments. The distribution of malignant and benign biopsies obtained from the 19 patients as well as the results of the different staining procedures are shown in Table I. CA125 was detected in 47 of 71 biopsies containing cancer obtained from nine patients (Figure 2). In one patient with an anaplastic ovarian cancer only two of 16 biopsies exhibited CA125. An apical staining pattern was observed without evidence for intracellular staining. In benign tissues CA125 could not usually be detected. Sometimes a diffuse staining pattern was observed in the excised abdominal laparotomy scar. The F(ab')2 fragments were detected in 34 of 71 biopsies from seven patients with cancerous tissue. The staining exhibited a diffuse, intracytoplasmic pattern, without
Table I Histological and immunohistochemical findings of 19 patients after immunoscintigraphy with F(ab')2 fragments of MAb OC125 Patient no. Carcinoma CA125 Biopsy F(ab')2 1 17 30 2 15 1 3 14 3 3 4 16 12 5 6 32 3 1 1 7 7 3 2 2 8 22 16 2 9 32 10 9 21 11 12 7 5 5 5 13 19 18 18 18 14 18 6 6 6 15 23 14 16 17 11 18 12 9 3 1 19 8 7 7 1 329 71 47 34 Biopsy = number of removed biopsies; carcinoma = number of biopsies containing cancer cells; CA125 = number of biopsies where CA125 was found; F(ab')2 = number of biopsies where F(ab')2 fragments of MAb OC125 were detected.
a
Figure 2 Apical reaction of the intact MAb OC125 with CA125 in papillary ovarian cancer ( x 250).
b
Figure 1 a Identical staining pattern of MAb OC125 (n) and incomplete MAb F(ab')2 OC125 (+) with CA125 on amniotic epithelium (note that the Fc fragment of the OC125 is missing in the right part of the drawing). b The reaction of incomplete MAb OC125 F(ab')2 with CA125 on amniotic epithelium ( x 250).
evidence of apical staining (Figure 3). In the remaining three patients with cancer no fragments could be demonstrated. In the benign tissue sections no staining of the fragments was observed.
Figure 3 Intracytoplasmic localization of MAb F(ab')2 OC125 in the same carcinoma as Figure 1. The slide was only incubated with the biotinylated rabbit antimouse antibody. Further staining was performed with the ABC detection system in the usual way.
HISTOCHEMISTRY OF OC125 AFTER IMMUNOSCINTIGRAPHY
Discussion For the detection of infused OC125 F(ab')2 fragments in different tissues it was necessary to determine whether the rabbit antimouse antibodies were able to recognize and bind to these fragments. For these experiments amniotic tissue was chosen, since extensive expression of CA125 in this tissue has already been documented (Kabawat et al., 1983). It could clearly be demonstrated that the rabbit antibodies could detect even the fragments of OC125. Since tissues were usually obtained several days after infusion of the OC125 fragments, it could not be excluded that radioactive decay would lead to a radiolysis of the antibodies which could disturb the immunohistochemical detection. However, even after storage of the radiolabelled antibodies for 14 days the same staining results were observed in amniotic epithelium. We therefore concluded that the immunohistochemical staining of the fragments in tissues obtained several days after antibody infusion was reliable.
69
For the immunohistochemical detection of the infused antibodies we omitted the usual first incubation with the OC125 antibody supplied with the kit. The results demonstrated that the infused antibodies could be detected in most of the cancer tissues, but not in benign tissues. The diffuse intracytoplasmic staining reaction indicated that the antibodies had crossed the cellular membranes of the cancer cells. It is not clear whether this is a result of prior diffusion of the cell membrane localized antigen-antibody complex into the cytoplasm or the reactions of the OC125 fragments with cytoplasmatic localized antigen or even endocytosis of antigen-antibody complexes built with free circulating antigens in blood vessels. Further studies are needed to substantiate these findings. However, the specific binding of OC125 to ovarian cancer tissue even after intravenous injection encourages the further application of this antibody for diagnostic and possibly therapeutic purposes.
References BAST, R.C., FEENEY, M., LAZARUS, H., NADLER, L.M., COLVIN,
GOLDENBERG, D.M., KIM, E.E. & DELAND, F.H. (1980). Radio-
R.B. & KNAPP, R.C. (1981). Reactivity of a monoclonal antibody with human ovarian carcinoma. J. Clin. Invest., 68, 1331. CHATAL, J.F., FUMOLEAU, P., SACCAVINI, J.C. & 7 others (1987). Scintigraphy of recurrences of gynecologic carcinomas. J. Nucl. Med., 28, 18.
immunodetection of cancer with radioactive antibodies to carcinoembryonic antigen. Cancer Res., 40, 2984.
DIETEL, M., ARPS, H.,
MOLLER-HAGEN, S., KLAPDOR, R. & HOFF-
MANN, L. (1986). Vergleichende Untersuchungen der Tumormarker CA125, CA19-9, CEA, CA17-IA und CA50 in Gewebsschnitten von Ovarialtumoren und den zugehorenden Patientenseren. In Klinische Relevanz Neuer Monoklonaler Antikorper, Greten, H. (ed.) p. 349. Thieme: Stuttgart. EPENETOS, A.A., BRITTON, K.E., MATHER, S. & 7 others (1982). Targeting of iodine 123-labelled tumour associated monoclonal antibodies to ovarian, breast and gastrointestinal tumours. Lancet, November 6, 999. FEISTEL, H., WOLF, F., JAGER, W., PATEROK, E.M. & LANG, N. (1988). Resection guided by antibodies (iodinized) - REGAJ.
Eine neue chirurgische Moglichkeit zur Lokalisierung von kleinsten Ovarialcarcinom-metastasen im AnschluB an Immunszintigraphie. In Radioactive Isotopes in Clinical Medicine and Research, H6fer, R. (ed.) p. 63. Schattauer: Stuttgart. GITSCH, E. & PATEISKY, N. (1987). Radioimmunszintigraphie beim Ovarialcarcinoma: theoretische Grundlagen und klinische Erfahrungen. Arch. Gynecol. Obstet., 242, 365. GOLDENBERG, D.M., DELAND, F., KIM, E. & 6 others (1978). Use of radiolabelled antibodies to carcinoembryonic antigen for the detection and localization of diverse cancers by external photoscanning. N. Engl. J. Med., 298, 1384.
HAISMA, H.J., MOSELEY, K.R., BATTAILE, A., GRIFFITH, T.C. &
KNAPP, R.C. (1988). Distribution and pharmacokinetics of radiolabelled monoclonal antibody OC-125 after intravenous and intraperitoneal administration in gynecologic tumnors. Am. J. Obstet. Gynecol., 159, 843. HSU, S.M., RAINE, L. & FRANGER, H. (1981). Use of avidin-biotinperoxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabelled antibody (PAP) procedures. J. Histochem. Cytochem., 29, 577. JAGER, W., FEISTEL, H., PATEROK, E.M., TULUSAN, A.H. & WOLF, F. (1987). Resection guided by antibodies iodinized (REGAJ): a surgical procedure to detect minor residues of ovarian cancer. J. Tumor Marker Oncology, 2, 215. JAGER, W., PATEROK, E. & FEISTEL, H. (1988). Resektion geleitet durch Antik6rper - REGAJ: Nachweis von Ovarial-carcinomen nach vorheriger Immunszintigraphie. TumorDiagnostik & Therapie, 9, 191. KABAWAT, S.E., BAST, R.C., BHAN, A.K., WELCH, W.R., KNAPP, R.C.
& COLVIN, R.B. (1983). Tissue distribution of a coelomicepithelium-related antigen recognized by the monoclonal antibody OC125. Int. J. Gynecol. Pathol., 2, 275. PIMM, M.V., PERKINS, A.C. & BALDWIN, R.W. (1985). Differences in tumour and normal tissue concentrations of iodine- and indiumlabelled monoclonal antibody. Eur. J. Nucl. Med., 11, 300. TAYLOR, C.R. (1986). Immunomicroscopy: A Diagnostic Toolfor the Surgical Pathologist. Saunders: Philadelphia.
X, 67 69 (I990), Br. 62, J. Suppl. Cancer (1990),
62,
Suppl.
X,
67
69
1990 Ltd., 1990~~~~~~~~
11" Macmillan Press © Macmillan Press Ltd.,
Immunohistochemical identification of CA125 and the F(ab'), fragments of the murine monoclonal antibody OC125 in ovarian cancer tissue G. Ronay', W. Jigerl, K. Weiss', H. Feistel2, F. Wolf2, A.H. Tulusan' & N. Lang' 'University of Erlangen, Department of Obstetrics and Gynecology, Universitdtsstrasse 21-23 and 2Department of Nuclear Medicine, Krankenhausstrasse 12, 8520 Erlangen, FRG. Summary Benign and malignant tissue samples of 19 ovarian cancer patients who received an intravenous infusion of radiolabelled F(ab')2 fragments of OC125 were examined for expression of CA125 and the cellular location of the injected antibodies. CA125 could be detected by immunohistochemistry in 47 of 71 biopsies containing cancer cells, but not in 258 biopsies with benign tissue. F(ab')2 fragments were found in 34 of these 47 biopsies. The diffuse intracytoplasmic staining pattern seemed to indicate that the injected antibodies had crossed the membranes of the cancer cells. Since the antibody could not be detected in benign tissues, further applications of OC125 for diagnostic and therapeutic purposes seems to be justified.
In 1981 Bast et al. described the murine monoclonal antibody (MAb) OC125 which bound to an antigen expressed on an ovarian cancer cell line. The antigen, which was coded CA125, was also detected in tissue sections of human epithelial ovarian cancers but not in the normal ovary (Kabawat et al., 1983; Dietel et al., 1986). It was therefore postulated that this specificity of the expression of CA125 could be used for the detection of ovarian cancerous tissues in patients. Radiolabelled fragments of OC125 were used for immunoscintigraphy and areas of increased radioactivity during scintigraphy were interpreted as specific accumulation of the antibodies to CA125 producing tissues, e.g. ovarian cancer. Thus preoperative information on tumour localization and metastatic spread could be achieved (Goldenberg et al., 1978, 1980; Epenetos et al., 1982; Pimm et al., 1985; Chatal et al., 1987; Gitsch & Pateisky, 1987). Recently intraoperative tracing has also been applied successfully (Jager et al., 1987, 1988). The distribution of the infused antibodies to different tissues has, however, so far only been described in terms of comparing the radioactivity localized in benign and malignant tissues (Haisma et al., 1988). In order to evaluate the binding of the injected OC125 antibody fragments we performed immunohistochemical staining procedures in different tissues obtained during surgery several days after immunoscintigraphy.
Patients, materials and methods
Antibody infusion In 19 ovarian cancer patients 1 mg of "3'I-labelled F(ab')2 fragments of the murine monoclonal antibody OC125 were infused at the Department of Nuclear Medicine, University of Erlangen as described previously (Feistel et al., 1988; Jiger et al., 1988). In two of these patients the antibodies were infused before primary surgery to a suspected ovarian cancer in order to evaluate if lymph node metastases could be detected. In the remaining 17 patients antibodies were infused prior to second-look operations to localize remaining tumour after chemotherapy or recurrence of the disease (Jager et al., 1988). Surgery Surgery was performed 7-13 days after infusion of the antibodies. During surgery biopsies were taken from different parts of the abdomen. Radioactivity was measured during surgery by a hand-held probe and biopsies which were Correspondence: G. Ronay. Supported by Istopen Diagnostik GmbH, Dreieich, FRG.
removed because of higher radioactivity compared with background were sampled (Jaiger et al., 1987). Further biopsies were resected from areas with background radioactivity as controls. All biopsies were fixed in Carnoy's solution (six parts ethanol 96%, three parts chloroform and one part glacial acetic acid) (Taylor, 1986) for 16-24 h according to the size of the tissue sample. Thereafter tissues were embedded in paraffin (Paraplast) at 56'C and stored until histological examination. Immunohistochemical staining of CA125 From each biopsy obtained, three tissue sections were prepared with a depth of 5 tm. One of the sections was used for routine haematoxylin staining procedures, while the remaining two sections were used for immunohistochemical staining according to the avidin-biotin-immunoperoxidase (ABC) technique (Hsu et al., 1981). Briefly, the samples were rehydrated for 3 x 10 min in xylol and 4 x 1 min in alcohol. Thereafter endogenous peroxidase was blocked by the addition of 0.3% H202-methanol solution for 20min. Since our previous studies seemed to demonstrate a partial destruction of the CA125 antigen by proteinases, incubation of tissues with trypsin and pronase was omitted during the study. To avoid non-specific binding of MAb OC125 via Fc receptors, the slides were treated before incubation with MAb with non-immunized sheep serum. After rinsing the tissue sections for 5 min in phosphate buffered saline (PBS), one of the two parallel sections was incubated with the undiluted murine MAb OC125 overnight at 4'C. During this time the parallel section remained stored until further processing in PBS. After this procedure a further incubation of both sections with the biotinylated rabbit antimouse antibody (dilution 1:50) and the streptavidine-biotin-peroxidase complex (dilution 1:100) (Amersham, FRG) was performed. The incubation period in each case was 30 min. The sections were rinsed for 3 x 5 min with PBS after every individual incubation step. Further reaction with 0.5% 3,3-diaminobenzidinetetrahydrochloride (DAB)/0.02% H202 solution resulted in the formation of a brown precipitate. Nuclear contrast staining was carried out using Mayer's haematoxylin. Immunohistochemical staining of the 0C125 F(ab')2 fragments For these experiments slices of human amniotic epithelium were used. The expression of CA125 in amniotic tissue was examined by immunohistochemistry using the monoclonal antibody OC125 as described above. To evaluate if the biotinylated rabbit antimouse antibodies also bound to the F(ab')2 fragments of OC125, amniotic epithelium was incubated with the OC125 F(ab')2 fragments
68
G. RONAY et al.
for 24 h. Thereafter the rabbit antimouse antibody was added to the tissue section and further staining was performed with the ABC detection system in the usual way. The OC125 F(ab')2 fragments were obtained from serum of patients, sampled 10 min after infusion of the antibodies. All experiments were repeated after storage of the serum for 14 days at 4°C. To exclude non-specific staining, amniotic epithelium was incubated with the ABC detection system without application of the OC125 or its fragments.
Results With the usual staining procedure the presence of the CA125 antigen in amniotic epithelium could be demonstrated. Using the OC125 fragments identical staining was observed, even when the stored sera were used (Figure 1). In the control experiment without the OC 125 antibodies no staining was detected. Three hundred and twenty-nine biopsies were examined for CA125 expression and presence of the F(ab')2 fragments. The distribution of malignant and benign biopsies obtained from the 19 patients as well as the results of the different staining procedures are shown in Table I. CA125 was detected in 47 of 71 biopsies containing cancer obtained from nine patients (Figure 2). In one patient with an anaplastic ovarian cancer only two of 16 biopsies exhibited CA125. An apical staining pattern was observed without evidence for intracellular staining. In benign tissues CA125 could not usually be detected. Sometimes a diffuse staining pattern was observed in the excised abdominal laparotomy scar. The F(ab')2 fragments were detected in 34 of 71 biopsies from seven patients with cancerous tissue. The staining exhibited a diffuse, intracytoplasmic pattern, without
Table I Histological and immunohistochemical findings of 19 patients after immunoscintigraphy with F(ab')2 fragments of MAb OC125 Patient no. Carcinoma CA125 Biopsy F(ab')2 1 17 30 2 15 1 3 14 3 3 4 16 12 5 6 32 3 1 1 7 7 3 2 2 8 22 16 2 9 32 10 9 21 11 12 7 5 5 5 13 19 18 18 18 14 18 6 6 6 15 23 14 16 17 11 18 12 9 3 1 19 8 7 7 1 329 71 47 34 Biopsy = number of removed biopsies; carcinoma = number of biopsies containing cancer cells; CA125 = number of biopsies where CA125 was found; F(ab')2 = number of biopsies where F(ab')2 fragments of MAb OC125 were detected.
a
Figure 2 Apical reaction of the intact MAb OC125 with CA125 in papillary ovarian cancer ( x 250).
b
Figure 1 a Identical staining pattern of MAb OC125 (n) and incomplete MAb F(ab')2 OC125 (+) with CA125 on amniotic epithelium (note that the Fc fragment of the OC125 is missing in the right part of the drawing). b The reaction of incomplete MAb OC125 F(ab')2 with CA125 on amniotic epithelium ( x 250).
evidence of apical staining (Figure 3). In the remaining three patients with cancer no fragments could be demonstrated. In the benign tissue sections no staining of the fragments was observed.
Figure 3 Intracytoplasmic localization of MAb F(ab')2 OC125 in the same carcinoma as Figure 1. The slide was only incubated with the biotinylated rabbit antimouse antibody. Further staining was performed with the ABC detection system in the usual way.
HISTOCHEMISTRY OF OC125 AFTER IMMUNOSCINTIGRAPHY
Discussion For the detection of infused OC125 F(ab')2 fragments in different tissues it was necessary to determine whether the rabbit antimouse antibodies were able to recognize and bind to these fragments. For these experiments amniotic tissue was chosen, since extensive expression of CA125 in this tissue has already been documented (Kabawat et al., 1983). It could clearly be demonstrated that the rabbit antibodies could detect even the fragments of OC125. Since tissues were usually obtained several days after infusion of the OC125 fragments, it could not be excluded that radioactive decay would lead to a radiolysis of the antibodies which could disturb the immunohistochemical detection. However, even after storage of the radiolabelled antibodies for 14 days the same staining results were observed in amniotic epithelium. We therefore concluded that the immunohistochemical staining of the fragments in tissues obtained several days after antibody infusion was reliable.
69
For the immunohistochemical detection of the infused antibodies we omitted the usual first incubation with the OC125 antibody supplied with the kit. The results demonstrated that the infused antibodies could be detected in most of the cancer tissues, but not in benign tissues. The diffuse intracytoplasmic staining reaction indicated that the antibodies had crossed the cellular membranes of the cancer cells. It is not clear whether this is a result of prior diffusion of the cell membrane localized antigen-antibody complex into the cytoplasm or the reactions of the OC125 fragments with cytoplasmatic localized antigen or even endocytosis of antigen-antibody complexes built with free circulating antigens in blood vessels. Further studies are needed to substantiate these findings. However, the specific binding of OC125 to ovarian cancer tissue even after intravenous injection encourages the further application of this antibody for diagnostic and possibly therapeutic purposes.
References BAST, R.C., FEENEY, M., LAZARUS, H., NADLER, L.M., COLVIN,
GOLDENBERG, D.M., KIM, E.E. & DELAND, F.H. (1980). Radio-
R.B. & KNAPP, R.C. (1981). Reactivity of a monoclonal antibody with human ovarian carcinoma. J. Clin. Invest., 68, 1331. CHATAL, J.F., FUMOLEAU, P., SACCAVINI, J.C. & 7 others (1987). Scintigraphy of recurrences of gynecologic carcinomas. J. Nucl. Med., 28, 18.
immunodetection of cancer with radioactive antibodies to carcinoembryonic antigen. Cancer Res., 40, 2984.
DIETEL, M., ARPS, H.,
MOLLER-HAGEN, S., KLAPDOR, R. & HOFF-
MANN, L. (1986). Vergleichende Untersuchungen der Tumormarker CA125, CA19-9, CEA, CA17-IA und CA50 in Gewebsschnitten von Ovarialtumoren und den zugehorenden Patientenseren. In Klinische Relevanz Neuer Monoklonaler Antikorper, Greten, H. (ed.) p. 349. Thieme: Stuttgart. EPENETOS, A.A., BRITTON, K.E., MATHER, S. & 7 others (1982). Targeting of iodine 123-labelled tumour associated monoclonal antibodies to ovarian, breast and gastrointestinal tumours. Lancet, November 6, 999. FEISTEL, H., WOLF, F., JAGER, W., PATEROK, E.M. & LANG, N. (1988). Resection guided by antibodies (iodinized) - REGAJ.
Eine neue chirurgische Moglichkeit zur Lokalisierung von kleinsten Ovarialcarcinom-metastasen im AnschluB an Immunszintigraphie. In Radioactive Isotopes in Clinical Medicine and Research, H6fer, R. (ed.) p. 63. Schattauer: Stuttgart. GITSCH, E. & PATEISKY, N. (1987). Radioimmunszintigraphie beim Ovarialcarcinoma: theoretische Grundlagen und klinische Erfahrungen. Arch. Gynecol. Obstet., 242, 365. GOLDENBERG, D.M., DELAND, F., KIM, E. & 6 others (1978). Use of radiolabelled antibodies to carcinoembryonic antigen for the detection and localization of diverse cancers by external photoscanning. N. Engl. J. Med., 298, 1384.
HAISMA, H.J., MOSELEY, K.R., BATTAILE, A., GRIFFITH, T.C. &
KNAPP, R.C. (1988). Distribution and pharmacokinetics of radiolabelled monoclonal antibody OC-125 after intravenous and intraperitoneal administration in gynecologic tumnors. Am. J. Obstet. Gynecol., 159, 843. HSU, S.M., RAINE, L. & FRANGER, H. (1981). Use of avidin-biotinperoxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabelled antibody (PAP) procedures. J. Histochem. Cytochem., 29, 577. JAGER, W., FEISTEL, H., PATEROK, E.M., TULUSAN, A.H. & WOLF, F. (1987). Resection guided by antibodies iodinized (REGAJ): a surgical procedure to detect minor residues of ovarian cancer. J. Tumor Marker Oncology, 2, 215. JAGER, W., PATEROK, E. & FEISTEL, H. (1988). Resektion geleitet durch Antik6rper - REGAJ: Nachweis von Ovarial-carcinomen nach vorheriger Immunszintigraphie. TumorDiagnostik & Therapie, 9, 191. KABAWAT, S.E., BAST, R.C., BHAN, A.K., WELCH, W.R., KNAPP, R.C.
& COLVIN, R.B. (1983). Tissue distribution of a coelomicepithelium-related antigen recognized by the monoclonal antibody OC125. Int. J. Gynecol. Pathol., 2, 275. PIMM, M.V., PERKINS, A.C. & BALDWIN, R.W. (1985). Differences in tumour and normal tissue concentrations of iodine- and indiumlabelled monoclonal antibody. Eur. J. Nucl. Med., 11, 300. TAYLOR, C.R. (1986). Immunomicroscopy: A Diagnostic Toolfor the Surgical Pathologist. Saunders: Philadelphia.
