CEA Expression Research Articles

The expression of CEA and HMFG antigens in ovarian clear-cell and endomemetroid carcinomas

The tumour antigen expression of ovarian and endometrial endometrioid carcinomas, ovarian clear-cell carcinomas as well as endometrial and cervical clear-cell carcinomas were immunohistochemically compared. Of special interest were potential differences between the endometrioid and clear-cell carcinomas of the ovary. The expression of CEA and CA 19-9 tumour antigens in all these tumour types was heterogeneous, with 10-20% of the cases being positive for CEA and 40-75% being positive for CA 19-9. In contrast, HMFG IIIC 12, a monoclonal antibody originally directed against human milk fat globule (HMFG) membrane antigens, invariably detected a corresponding antigen on every case of these tumour types. Another HMFG antibody, SM IF 3, on the other hand, detected antigenic material on all clear-cell tumour types, but only rarely on endometrioid tumours of the ovary or endometrium. While HMFG IIIC 12 detects an antigen present on all ovarian, endometrial and mammary carcinomas, antibody SM IF 3 thus appears to be more restricted in its staining patterns. Our results with both of these antibodies indicate that ovarian clear-cell carcinomas and ovarian endometrioid carcinomas have antigenic differences, which provides further evidence that they belong to different tumour entities.

胆嚢癌の壁深達度と核ＤＮＡ量およびＣＥＡ発現との関連性

胆嚢癌の壁深達度と核ＤＮＡ量およびＣＥＡ発現との関連性

Aberrant expression of intestinal mucin antigens associated with colorectal carcinoma defined by a panel of monoclonal antibodies

Small intestine mucin antigen (SIMA) is an oncofoetal antigen for the colon and is distinct from the normal large intestinal mucin antigen (LIMA). In the present study, a panel of anti-SIMA and anti-LIMA monoclonal antibodies (MAb) was used to charaterise altered mucin expression in colorectal adenocarcinomas, by immunohistochemistry and quantitative immunoassays of tissue extracts. These results are compared with CEA expression and correlated with various clinicopathological indices. All mucin MAb reacted with a high proportion of the 100 colon cancers of every stage, histological type (including non-mucinous cancers), differentiation, site, or size. Inappropriate SIMA production was detected by either anti-SIMA MAb 4D3 or 4A1, even in 85% of early stage cancers. MAb 4D3 reacted with a higher proportion of cancers of smaller size and better differentiation. At the subcellular level, both anti-SIMA MAb showed reactivity typical of normal mucin, i.e., goblet cell and extracellular mucin. The normal colonic antigen, LIMA, was also detectable in the majority of cases, but quantitatively overproduced in some cases and reduced in others. However, in contrast to SIMA, LIMA was detected in predominantly undifferentiated cancer cells but not in goblet cells. Heterogeneity of MAb reactivity between cases and complementarity within each cancer was frequently observed. Mucin reactive with at least one of the MAb was detected in all of the CEA-negative cancers. A high rate of inappropriate SIMA expression was also detected in the perineoplastic transitional mucosa (88%, c.f. CEA, 35%) and adjacent, morphologically normal mucosa (80% c.f. CEA, 24%), indicating biochemical changes similar to the cancer. This panel of anti-mucin MAb demonstrated altered mucin glycoprotein metabolism associated with the development and progression of most colorectal cancers, which emphasises their utility as indicators of neoplastic change in the colon, and their superiority to CEA.

Establishment and characterisation of human carcinoembryonic antigen transgenic mice

We have produced human CEA transgenic mice which were found to express CEA mRNA in all tissues. By immunoblot analysis using anti-CEA polyclonal antibody, we also detected CEA protein in all tissues. However, the molecular size of CEA in the brain was different from that in other tissues, although the mRNA size was same and no deletion nor rearrangement was detected at the DNA level. Immunohistochemical analysis of the lung and the colon showed that the expression sites were the bronchial epithelial cells of the lung and the columnar epithelial cells of the colon. Interestingly, the expression of CEA protein in the transgenic mice was polarised to the luminal side of epithelial cells similar to the normal CEA expression in human tissues. We also detected cell surface expression of human CEA on thymocytes and spleen cells and CEA expression was greatly reduced by the phosphatidylinositol-specific phospholipase C (PI-PLC) treatment.

The immunoscintigraphic use of Tc‐99m‐labelled monoclonal anti‐CEA antibodies (BW 431/26) in patients with suspected primary, recurrent and metastatic breast cancer

The discrepancy between serum CEA levels and CEA tissue expression in patients with breast cancer is well known. Whereas immunohistochemistry shows positive CEA expression in 70-90%, the serum CEA levels are often within the normal range. We performed immunoscintigraphy and SPECT with a Tc-99m labelled anti-CEA monoclonal antibody (MAb BW 431/26) in 46 women with suspected breast cancer or recurrence. The results of anti-CEA immunoscintigraphy, mammography, serum CEA levels and immunohistochemistry were evaluated according to the histology of the tumor. Histology verified breast cancer or recurrence (pT1 [n = 7], pT2 [n = 17], pT3 [n = 3], pT4 [n = 3]) in 30 out of 46 patients; benign breast disease such as fibrocystic disease, fibroadenoma, fatty necrosis or chronic mastitis was responsible for suspicious mammographic findings in 16 patients. Immuno-SPECT showed 25 true-positive, 5 false-negative, 11 true-negative and 5 false-positive findings (sensitivity 83%, specificity 69%). Anti-CEA immuno-SPECT of 2 patients with bone metastasis showed all lesions previously detected by bone scintigraphy to be CEA-expressing metastases. In contrast, serum CEA levels were slightly elevated in only 5 out of 30 patients with histologically verified breast cancer (sensitivity 17%). The results of immuno-histochemistry were surprising; tissue CEA expression could be demonstrated in only 5 patients with breast cancer. According to our experiences with this Tc-99m labelled anti-CEA MAb, immuno-SPECT is a suitable additional method for the diagnosis of breast cancer and especially of recurrence. Pre-operative serum CEA levels give no support for the differentiation between benign and malignant breast tumors.

Malignant Islet Cell Tumor of the Pancreas with Multiple Hormone Production and Expression of CEA and CA19‐9

An autopsy case of malignant islet cell tumor of the pancreas is presented. The patient, a 64-year-old woman showed severe hypoglycemia as the initial symptom, and hyperinsulinemia was demonstrated by laboratory examinations. Metastatic tumors in the liver were found by abdominal computed tomography. Autopsy revealed a tumor measuring 6.5 x 3 x 2 cm occupying the pancreas from the body to the tail. From the results of histological and immunohistochemical studies, this was diagnosed as a malignant islet cell tumor producing multiple hormones such as insulin, glucagon, somatostatin and pancreatic polypeptide, as well as expressing the tumor-related antigens CEA and CA19-9. These findings suggested that the tumor cells showed differentiation to both endocrine cells and pancreatic duct cells.

Immunohistochemical detection of CEA and P-glycoprotein (P-gp) in small cell lung cancer (SCLC): With special reference to the tissue expression of CEA and response to chemotherapy: Yoshihiko Segawa, Taisuke Ohnoshi, Shunkichi Hiraki, Hiroshi Ueoka, Kazuyo Miyatake and Ikuro Kimura Department of Medicine, Okayama University Medical School, Okayama 700, Japan

Immunohistochemical detection of CEA and P-glycoprotein (P-gp) in small cell lung cancer (SCLC): With special reference to the tissue expression of CEA and response to chemotherapy: Yoshihiko Segawa, Taisuke Ohnoshi, Shunkichi Hiraki, Hiroshi Ueoka, Kazuyo Miyatake and Ikuro Kimura Department of Medicine, Okayama University Medical School, Okayama 700, Japan

Radioimmunolocalization using the radiolabeled monoclonal anti-cea antibody 11-285-14 lacks diagnostic utility for the differentiation of pancreatic and bile duct cancer from chronic pancreatitis and sclerosing cholangitis

CEA expression was determined by immunohistochemistry in paraffin sections from cases of pancreatic cancer(n = 30), chronic pancreatitis(n = 10), cholangiocarcinoma(n = 12), and sclerosing cholangitis (n-4) using a new anti-CEA monoclonal antibody (11-285-14). CEA was expressed in 77% of pancreatic cancers and 80% of cholangiocarcinomas, but was not detected in any of the cases of sclerosing cholangitis. Although 60% of chronic pancreatitic tissues were positive, staining intensity was markedly reduced compared to cancers. Twenty-five patients with these conditions were therefore studied using radioimmunolocalization. Eleven of 12 pancreatic cancers were positively imaged, as were all three biliary tumors. However, positive scans were also obtained in 50% of the cases of chronic pancreatitis and sclerosing cholangitis. The overall sensitivity (68%) and specificity (63%) of radioimmunolocalization using the 11-285-14 antibody were not high enough to justify its use for diagnosis.

Mutation of the promoter and LexA binding sites of cea, the gene encoding colicin E1.

Three mutations were introduced into the cea promoter using oligonucleotide directed mutagenesis. The resulting mutant promoter has the Escherichia coli consensus sequences at its - 35 and - 10 positions, separated by the optimal spacing. In addition, a plasmid with a mutation in one of the two LexA repressor binding sites in the cea regulatory region was isolated that decreases homology with the consensus LexA binding site. The effects of these mutations on cea expression were studied in cea-lacZ protein fusions. The promoter-up mutant, when present in a multicopy plasmid, showed a shorter induction lag when compared to the wild-type cea gene, and there was less of an effect of the catabolite repression system on cea expression. However, when present in a single copy in the bacterial chromosome, catabolite repression and an induction delay were observed, despite the increased strength of the promoter. The operator mutant showed a slightly higher basal level of expression, but was still repressible. Induction occurred with a shortened lag period, but the effects were not as great as with the promoter mutant. These results support the idea that tight repression by LexA contributes to the delay in cea induction.

Human gastric cancer with expression of CEA, ER, E2, EGF and EGF-R

Human gastric cancer with expression of CEA, ER, E2, EGF and EGF-R

Epithelial cell markers and proliferating cells in odontogenic jaw cysts

The expression of keratins, CEA, EMA, and rat liver antigen (RLA) and the presence of Ki67+ proliferating cells were studied in the epithelial linings of 50 odontogenic cysts using an indirect immunoperoxidase method on acetone-fixed frozen sections. All cysts were positive with monoclonal antibodies of broad keratin specificity (CK1, AE1-3), and between 40 and 100 per cent of epithelial cells expressed keratins 13 and 19. Keratins 7, 8, and 18 were rarely expressed although surface cells in areas of mucous metaplasia often expressed keratins 7 and 18. Expression of keratin 10/11 was related to the presence of a well-ordered epithelial lining and was detected in isolated cells in 4/32 non-keratinizing cysts and in the upper suprabasal cell layers of 17/18 keratocysts. Although CEA, EMA, and RLA were detected in the epithelium of all specimens, the pattern of expression of CEA and EMA differed between cyst types. Ki67+ proliferating cells were most prevalent in keratocyst epithelia, where they were usually found within lower suprabasal layers which were negative or weakly positive for keratins 10/11 and 13. These results indicate differences in keratin, CEA, and EMA expression between cyst types which appear to be dependent on epithelial differentiation/structure rather than cyst type or histogenesis. Although these differences may not be of diagnostic significance, the consistent expression of both keratins 13 and 19 may provide a useful marker of odontogenic epithelium in general.

Immunohistochemical Studies on Cystic Pancreatic Neoplasms

For the treatment of cystic changes of the pancreas, it is essential to distinguish cysts and pseudocysts from neoplasm. Since clinical parameters are usually not characteristic, only histologic and cytologic analyses will prove a diagnosis. The immunohistochemical characteristics of microcystic adenoma, mucinous cystic neoplasia in comparison to solid cystic pancreatic tumor, ductal carcinomas, and endocrine tumors, are studied with a panel of markers as well as enzyme, epithelial, neuroendocrine and pancreatic hormonal markers. The immunohistochemical results with diffuse cytoplasmatic expression of CEA and epithelial markers may be helpful in the exploration of carcinomatous transformed tissue parts within mucinous cystic pancreatic neoplasias, and are therefore of significance for surgical therapy.

Expression of CEA, CA125, CA19-9 and human milk fat globule membrane antigen in ovarian tumours.

The expression of five different antigens in ovarian tumours was studied by means of an immunohistochemical test with anti-CEA, HMFG1 and HMFG2, NS19-9 and OC125 antibodies. Considerable variation was noted not only between different histological types and between tumours of one type but also between areas in a single tumour. HMFG1 and HMFG2 were the most reactive of all the antibodies; NS19-9 and OC125 were expressed by different populations of cells. It is concluded that specific combinations of antibodies are more effective both for the monitoring of ovarian cancer as well as for immunodiagnosis and treatment, than any single one used.

Biochemical tissue markers of human colorectal carcinoma.

The potential therapeutic effects of differentiating agents on leukemic and solid tumor cells are being evaluated worldwide. These effects can be followed by morphologic as well as biochemical parameters. The enzymatic profile of four enzymes and the level of carcinoembryonic antigen were studied in 24 human colorectal carcinoma specimens and their adjacent uninvolved mucosa. The enzymes studied were thymidine kinase and 6-phosphogluconate dehydrogenase as markers of proliferation, and alkaline phosphatase and gamma-glutamyl transpeptidase as markers of differentiation. A consistent finding was a marked increase in the activities of thymidine kinase and 6-phosphogluconate dehydrogenase in the tumor cells as compared with the adjacent normal mucosa. The activity of gamma-glutamyl transpeptidase was not significantly different between tumor and uninvolved colon tissue. Alkaline phosphatase activity was markedly reduced in the tumor specimens. A relationship between the degree of differentiation and the degree of penetration and CEA expression was demonstrated in the tumor specimens as well as in their surrounding uninvolved mucosa.

Temporal control of colicin E1 induction.

The expression of the gene encoding colicin E1, cea, was studied in Escherichia coli by using cea-lacZ gene fusions. Expression of the fusions showed the same characteristics as those of the wild-type cea gene: induction by treatments that damage DNA and regulation by the SOS response, sensitivity to catabolite repression, and a low basal level of expression, despite the presence of the fusion in a multicopy plasmid. Induction of expression by DNA-damaging treatments was found to differ from other genes involved in the SOS response (exemplified by recA), in that higher levels of DNA damage were required and expression occurred only after a pronounced delay. The delay in expression following an inducing treatment was more pronounced under conditions of catabolite repression, indicating that the cyclic AMP-cyclic AMP receptor protein complex may play a role in induction. These observations also suggest a biological rationale for the control of cea expression by the SOS response and the cyclic AMP-cyclic AMP receptor protein catabolite repression system.

Quantitation of MHC antigen expression on colorectal tumours and its association with tumour progression.

A flow cytometric technique has been established for accurately quantitating the cell surface density of MHC antigens and the percentage of cells expressing MHC antigens in 38 colorectal tumours. Thirty-four percent of tumours were partially or completely negative for HLA-ABC antigen expression. Although the quantity of HLA-ABC antigens varied widely, there was no correlation between the density of HLA-ABC antigens, or the percentage of cells expressing these antigens and clinicopathological stage. Fifty percent of the colorectal tumours expressed HLA-DR with varying antigen densities. All of the poorly differentiated tumours expressed HLA-DR but there was no correlation between expression of HLA-DR and clinicopathological stage. The aneuploid tumours expressed more HLA-ABC and HLA-DR antigens on a higher percentage of cells than the diploid tumours. Abnormal expression of the tumour associated antigens CEA, Y haptenic blood group and 791T p72 also correlated with expression of HLA-ABC and HLA-DR antigens on colorectal tumours. The majority of early derived in vitro dividing cells failed to express both HLA-ABC and HLA-DR antigens although they expressed high levels of tumour associated antigens. If there is a correlation between in vitro and in vivo growth perhaps tumours are maintained and seeded by MHC antigen negative cells.

An experimental model for pharmacokinetic studies of monoclonal antibodies in human colonic cancer.

An experimental model consisting of athymic rats carrying human colonic tumours from cell line LS 174T in both hind legs was used. 125I-labelled anti-carcinoembryonic antigen (anti-CEA) monoclonal antibodies were injected intra-arterially (i.a.), either alone (21 rats) or together with degradable starch microspheres (6 rats). As a control, an irrelevant antibody was injected i.a., alone (6 rats) or together with microspheres (3 rats). An intra-arterial injection was given on the side bearing one tumour in each rat, while the contralateral tumour served as an 'intravenous' control. The rats were submitted to external gamma measurements daily for four days. On the fourth day they were killed and pieces from the tumours and from various organs were examined by in vitro measurements. The results indicate strong expression of CEA in LS 174T cells grafted to athymic rats. No lasting enhancement of the tumour uptake was achieved by intra-arterial injection of antibodies as compared with the control tumours.

Prognostic significance of CEA immunoreactivity patterns in large bowel carcinoma tissue.

In order to determine the clinical value of CEA detection in large bowel cancer tissue the patterns rather than the intensity of immunoreactivity of CEA reactive antibodies were analyzed in 312 large bowel cancer patients especially in relation to patient survival. CEA immunoreactivity appeared to be distinguishable into a predominantly apical/cytoplasmic and a predominantly membranous pattern. Twenty-four (7.7%) tumours were found to be CEA negative or only focally positive. Two hundred and eighty-three (90.7%) of the carcinomas showed a predominantly apical/cytoplasmic immunoreactivity pattern, whereas 5 (1.6%) of the tumours revealed mostly membranous CEA immunoreactivity. CEA negative or focally positive carcinomas and CEA positive tumours with membranous immunoreactivity were significantly more often observed in the group of poorly differentiated carcinomas (P greater than 0.001), but showed no significant correlation with stage of tumour extension (P = 0.11). Also, these carcinomas demonstrated a more aggressive course in patients compared to CEA positive tumours with an apical/cytoplasmic CEA expression pattern. We, therefore, conclude that determination of the pattern of CEA immunoreactivity in large bowel cancer tissue may enable the detection of subgroups of patients with a poor prognosis.

Combined immunohistochemical studies and proliferation kinetic on urothelial atypias and carcinomas in the same material

Combined immunohistochemical studies and p ro l i f e ra t i on k i netic were performed on ident ica l s l ides of atypias and carcinomas of the urinary bladder mucosa. Those studies were designed to invest igate a possible corre la t ion between h is to log ica l , immunohistochemical, and ce l l k ine t ic character is t ics. 71 t issue specimens that had already been treated autoradiographical ly , were also examined immunhistochemica l ly . Labell ing index inreases from 1,4% in s l i gh t to 20% in marked uro the l ia l atypia. CEA reaction in s l i gh t atypia is s l i gh t or moderate, s l igh t , moderate or d i s t i nc t in moderate atypia, and moderate to d i s t i nc t in carcinoma in s i tu . TPA always shows moderate to d i s t i nc t reactions. Cell k i ne t i ca l l y , uro the l ia l carcinomas, y ie ld s imi lar graduations. They were pos i t ive for CEA in 70% and for TPA in 100%. In G 0 and G I carcinomas negative and s l i g h t l y pos i t ive reactions predominate, poorly d i f f e rent iated lesions y ie ld predominantly d i s t i nc t reactions. In a l l grades, TPA ranges from s l i gh t to d i s t i n c t l y pos i t ive. As in ce l l k inet ic analyses, there is a re la t ion ship between d i f f e ren t i a t i on grade and stage for CEA expression. This does not apply fo r TPA. The results permit us to draw conclusions on the d i f f e rent b io log ica l and histogenet ic behavior of u ro the l ia l carcinomas and atypias. There are undoubtedly differences in the behavior of pap i l l a r y exophytical and so l id invasive carcinomas in terms of both ce l l k inet ics and immunohistochemistry.

Monoclonal antibodies against CEA-related components discriminate between pancreatic duct type carcinomas and nonneoplastic duct lesions as well as nonduct type neoplasias.

The expression of CEA and related antigens in formalin-fixed paraffin-embedded tissues of normal pancreas and different pancreatic neoplasms was studied immunocytochemically using three monoclonal antibodies (MAbs) recognizing different epitopes on CEA and related antigens. Additionally, a number of extrapancreatic malignancies were tested. The epitope recognized by MAb 250 (present on CEA and NCA 95) was expressed in all but one pancreatic ductal adenocarcinoma and ampullary carcinoma (42/43). The MAb 431 defined epitope (present only on CEA) was less frequently found (27/43). MAb 374, defining an epitope on CEA, NCA 95 and NCA 55 proved to be nearly as sensitive as MAb 250, but also reacted with normal duct epithelium. In contrast, MAb 250 and MAb 431 discriminated clearly between reactive duct lesions and malignant duct changes. Moreover, these MAbs differentiated between pancreatic duct carcinomas and nonduct type carcinomas as well as benign pancreatic tumours. In duct type carcinomas, the strongest staining was observed in well differentiated tumours. No discrimination was possible between pancreatic carcinomas and other adenocarcinomas of the gastrointestinal tract nor between most of the lung carcinomas and some other malignancies, specified below. MAb 250 and MAb 431 failed to react with hepatocellular carcinomas, renal cell carcinomas, carcinoids, sarcomas and melanomas. The findings suggest that paraffin-embedded tissues of pancreatic duct type carcinomas, in contrast to nonduct type tumours and normal ducts, are distinguished by the presence of a CEA and NCA 95 related epitope.