Serum levels and clinical evaluation of CA 19-9 in various diseases

We evaluated the clinical significance of serum CA 19-9 in various cancers, benign diseases and healthy controls. The percentage of positive cases for CA 19-9 (higher than 37 U/ml) was 83% in pancreas cancer, 78% in biliary tract cancer and 75% in urinary tract cancer. Benign diseases showed a low frequency of positive cases for CA 19-9, and their serum levels of CA 19-9 were low. Benign diseases with high serum levels of CA 19-9 were rarely seen, but they were easy to differentiate from malignant tumors by simultaneous examination of other tumor markers. In cancer patients with high serum levels of CA 19-9, CA 19-9 showed a weak positive correlation to CEA and ferritin.     

Clinical evaluation of radioimmunoscintigraphy using a I-131 anti CEA and CA 19-9 monoclonal antibodies cocktail (IMACIS-1)

The clinical usefulness of a I-131 anti CEA F(ab')2 MoAB and CA 19-9 F(ab')2 MoAB cocktail(IMACIS-1) for the localization of a cancer has been studied in 35 patients (Lung Ca. 10, Pancreas Ca. 7, Gastric Ca. 6, Colon Ca. 5, Hepatoma 2, Cholangio Ca. 2, Esophageal Ca. 1, Uterus Ca. 1, and, Retroperitoneal tumor 1). Of these 28 patients (80%) had an abnormal accumulation of radioactivity in the tumor sites (Lung Ca. 5, Pancreas Ca. 7, Gastric Ca. 6, Colon Ca. 5, Hepatoma 2, Esophageal Ca. 1, Uterus Ca. 1, and, Retroperitoneal tumor 1). Eighteen cases (56.3%) in 32 cases in which the primary site was known showed an abnormal radioactive accumulation and, similarly, 20 metastatic sites (71.4%) in 16 cases with 28 metastatic sites also showed an abnormal accumulation of radioimmunoscintigraphy showed no correlation with the serum CEA or CA 19-9 concentration. Further, no side effect was observed.     

Radiosensitizer conjugation to the carcinoma 19-9 monoclonal antibody

Misonidazole was covalently conjugated (3-68 mol drug/mol antibody) to 19-9 monoclonal antibody directed against a colorectal carcinoma tumor-associated antigen as a method for targeting radiosensitizing agents. This attachment was accomplished by the mixed anhydride method using the hemisuccinate derivative of misonidazole. Evaluation of conjugates in vitro shows a loss of antibody binding activity with increasing loading levels; however, significant binding activity is retained even at relatively high sensitizer/antibody ratios. This observation was consistent in three binding assays: a competitive radioimmunoassay; an enzyme immunoassay; and an affinity column assay. From these studies, it was concluded that the optimal loading factor for misonidazole-antibody conjugates, when it is used for immunochemotherapy lies between 8 and 15. In vitro release studies indicated that conjugates are hydrolytically stable (t1/2 = 4 days) under physiological conditions.     

Initial clinical evaluation of two murine IgG2a monoclonal antibodies for immunotherapy of gastrointestinal carcinoma

Eleven patients with advanced gastrointestinal (GI) carcinoma were entered in Phase I initial clinical trials with IgG2a antiGI carcinoma monoclonal antibodies (MAbs) GA733 (five patients) or CO19-9 (six patients). Infusion of MAb GA733 in doses greater than 30 mg was accompanied by mild and short-lasting GI toxicity. Infused MAb GA733 was bound to each patient's tumor tissue in vivo. MAb circulated in the blood for 10-25 days. All patients developed anti-mouse antibodies between 15 and 60 days post infusion. Furthermore, all but one patient raised anti-idiotypic antibodies against MAb GA733. Following administration of 10-600 mg of MAb CO19-9, no immediate or delayed toxicity symptoms were noted. Binding of infused MAb CO19-9 to tumor cells in vivo could not be detected in any of the six patients studied. The MAb circulated in the bloodstream between 5 and 12 days. Human anti-mouse antibody was detected in sera of three patients. None of the eleven patients treated with either MAb had anti-tumor responses in this Phase I clinical trial. The strong binding reactivity of MAb GA733 to tumors in vivo suggests the use of this MAb in cancer patients with less tumor burden to determine the tumoricidal efficacy of this antibody.     

Diagnostic and therapeutic utility of monoclonal antibodies in urologic oncology

Remarkable advances in the treatment of urologic malignancies have recently been made. Monoclonal antibodies selective for a variety of normal and malignant urologic tissues have been useful in defining normal antigens and tumor-associated antigens and have potential as diagnostic and immunotherapeutic agents. In renal cancer, monoclonal antibodies can define serum markers, radiolabel tumor xenografts, and assist in specific tissue diagnosis. Additionally, there is potential for these antibodies either alone or as conjugates to localize and kill tumors. Monoclonal antibodies to bladder cancer associated antigens are able to demonstrate differential antigen expression on superficial versus invasive tumors, to refine urinary cytologic diagnosis of bladder cancer, and to predict invasive recurrence of superficial cancer. Monoclonal antibodies have localized bladder tumor xenografts and can inhibit tumor growth when conjugated to radioisotopes or toxins. In prostate cancer monoclonal antibodies to prostate antigens are not usually tumor specific. Monoclonal antibodies to prostate antigen (PA) and prostatic acid phosphatase (PAP) are able to localize prostate cancer metastases. Chemotherapy-conjugated anti-PAP monoclonal antibodies have demonstrable inhibition on human prostate cancer xenografted tumor growth. Monoclonal antibodies have defined normal and tumor-associated antigens in urologic cancers and are expected to be useful in immunodiagnosis and cancer therapy in the near future.     

Clinical utility of radiolabeled monoclonal antibodies in prostate cancer

Prostate cancer represents an ideal target for radioimmunotherapy based on the pattern of spread, including bone marrow and lymph nodes, sites that typically receive high levels of circulating antibody, and the small volume of disease, ideally suited for antibody delivery and antigen access. This review explores possible antibody targets in prostate cancer and focuses on the potential role for radioimmunotherapy by highlighting several clinical trials involving radiolabeled anti-prostatespecific membrane antigen monoclonal antibody J591. Prostate-specific membrane antigen, a highly prostate-restricted transmembrane glycoprotein with increased expression in high-grade, metastatic, and hormone-refractory disease, represents an ideal target for monoclonal antibody therapy in prostate cancer. Radiolabeled anti-prostate-specific membrane antigen monoclonal antibody J591 trials using the radiometals yttrium-90 and lutetium-177 have demonstrated manageable myelotoxicity, no significant nonhematologic toxicity, excellent targeting of soft-tissue and bone metastases, and preliminary efficacy including prostate-specific antigen and measurable disease responses. Additional studies are under way to better define the activity of radiolabeled antibody therapy as well as the role for fractionated therapy and combination approaches with taxane-based chemotherapy.     

The utility of monoclonal antibodies in the imaging of prostate cancer

Monoclonal antibodies (mAbs) to prostate-specific antigens, such as PSMA, have great potential as diagnostic and therapeutic tools in the management of advanced prostate cancer. PSMA is a very attractive target for mAb-based imaging. It is expressed by virtually all prostate cancers and its expression is further increased in poorly differentiated, metastatic, and hormone-refractory carcinomas. The ProstaScint scan (Cytogen, Princeton, NJ), based on the mAb 7E11-C5.3, is currently approved for the imaging of prostate cancer in soft tissue but is not approved for imaging bone metastases. It appears superior to conventional imaging studies for soft-tissue disease but has limitations attributed to its intracellular binding site on PSMA. Overcoming this limitation, new mAbs to the extracellular domain of PSMA have been developed. The radioisotopes, (111)Indium, (90)Yttrium, and (177)Lutetium have been conjugated to one such mAb, J591. Radioimmunoscintigraphy with this immunoconjugate has demonstrated excellent tumor targeting of prostate cancer sites not only in soft tissue but also in bone.     

A clinical evaluation of carbohydrate antigen 19-9 and carcinoembryonic antigen in patients with pancreatic carcinoma

We have studied serum carbohydrate antigen 19-9 (CA 19-9) and carcinoembryonic antigen (CEA) in 221 persons to assess their usefulness in the diagnosis of pancreatic carcinoma. Although serum CA 19-9 and CEA in all healthy controls were within normal limits, the positive ratings of serum CA 19-9 and CEA in all benign disease were 9.8% and 18.1%, respectively. Sensitivity of serum CA 19-9 for pancreatic carcinoma was 70.5%, which was higher than that found in healthy controls, benign disease, and other malignant disease except biliary carcinoma; but sensitivity of serum CEA levels (67.7%) was not different from that seen in malignant disease. Three of 34 patients (8.8%) with pancreatic carcinoma who had a above-normal levels of serum CA 19-9 but not serum CEA were resectable. Although there was no correlation between serum CA 19-9 and CEA, advanced stages of pancreatic, gastric, and colorectal carcinoma tend to show high serum CA 19-9 and CEA, but no statistical differences were observed in relation to the stages of these carcinomas. Comparative studies of serum CA 19-9 and CEA for sensitivity and the predictive value of true positive and negative results for detecting pancreatic, gastric, and colorectal carcinoma showed that serum CA 19-9 has significantly higher sensitivity and predictive value of true positive results for pancreatic carcinoma than for gastric and colorectal carcinoma (P less than 0.05). However, serum CEA measurements did not show any difference between these carcinomas, and the highest predictive value of a true negative result for excluding pancreatic carcinoma was also observed in serum CA 19-9. These results indicate that although the CA 19-9 assay is not specific for pancreatic carcinoma, it is more useful adjunct method for diagnosing pancreatic carcinoma, possibly in resectable stages.     

Initial clinical evaluation of an immunoradiometric assay for CA 19-9 using the NCI serum bank

More than 1,600 coded sera obtained from blood donors and the NCI/Mayo Clinic Serum Bank were analyzed with an improved immunoradiometric assay for the carbohydrate antigenic determinant, CA 19-9. Results indicated that CA 19-9 is elevated in a large fraction of sera (67%) from patients with advanced adenocarcinomas of the upper gastrointestinal (GI) tract, including those with pancreatic, hepatobiliary and gastric carcinomas. Several of these sera had CA 19-9 exceeding 300,000 U/ml. A smaller fraction (18%) of patients with carcinomas of the large bowel had elevated serum CA 19-9 levels, the majority among patients with metastatic disease. In contrast, none of the healthy donors from the serum bank and only 4 of 1,023 of the blood donor specimens (0.4%) had CA 19-9 levels greater than or equal to 40 U/ml. Three of 235 sera (1.3%) from benign disease patients had levels of CA 19-9 in excess of 40 U/ml. These data suggest that the improved CA 19-9 immunoradiometric assay may have clinical utility as a diagnostic adjunct for adenocarcinoma of the upper GI tract and that the assay also may have some value in monitoring patients with advancing colorectal carcinoma, particularly in combination with CEA determinations. Rigorous prospective clinical trials will be necessary to verify these hypotheses.     

The clinical utility of serum CA 19-9 in the diagnosis,prognosis and management of pancreatic adenocarcinoma: An evidence based appraisal

Background

Serum carbohydrate antigen (CA 19-9) is the most common tumor marker assessed in pancreatic cancer patients; nevertheless few articles have comprehensively evaluated the evidence for its utility in pancreatic cancer management.

Methods

Literature search was performed using Medline with keywords "pancreatic cancer", "tumor markers", "CA 19-9", "diagnosis", "screening", "prognosis", "resectability" and "recurrence". All English language articles pertaining to the role of CA 19-9 in pancreatic cancer were critically analyzed to determine its utility as a biomarker for pancreatic cancer.

Results

Serum CA 19-9 is the most extensively validated pancreatic cancer biomarker with multiple clinical applications. CA 19-9 serum levels have a sensitivity and specificity of 79-81% and 82-90% respectively for the diagnosis of pancreatic cancer in symptomatic patients; but are not useful as a screening marker because of low positive predictive value (0.5-0.9%). Pre-operative CA 19-9 serum levels provide useful prognostic information as patients with normal levels (<37 U/mL) have a prolonged median survival (32-36 months) compared to patients with elevated levels (>37 U/mL) (12-15 months). A CA 19-9 serum level of <100 U/mL implies likely resectable disease whereas levels >100 U/mL suggest unresectablity or metastatic disease. Normalization or a decrease in post-operative CA 19-9 serum levels by ≥20-50% from baseline following surgical resection or chemotherapy is associated with prolonged survival compared to failure of CA 19-9 serum levels to normalize or an increase. Important limitations to CA 19-9 serum level evaluation in pancreatic cancer include poor sensitivity, false negative results in Lewis negative phenotype (5-10%) and increased false positivity in the presence of obstructive jaundice (10-60%).

Conclusions

CA 19-9 is the most extensively studied and validated serum biomarker for the diagnosis of pancreatic cancer in symptomatic patients. CA 19-9 serum levels can provide important information with regards to prognosis, overall survival, and response to chemotherapy as well as predict post-operative recurrence. However, non-specific expression in several benign and malignant diseases, false negative results in Lewis negative genotype and an increased false positive results in the presence of obstructive jaundice severely limit the universal applicability of serum CA 19-9 levels in pancreatic cancer management.Key Words : Pancreatic cancer, tumor markers, CA 19-9, diagnosis, screening, prognosis, resectability, recurrence     

Characterisation and clinical usefulness of CA130 antigen recognised by monoclonal antibodies, 130-22 and 145-9, in ovarian cancers.

A new cancer-associated antigen CA130, recognised by two monoclonal antibodies (moABs) 130-22 and 145-9, was often found to be present at high levels in the sera of patients with ovarian cancer. There was a strong correlation between CA130 and CA125 values. The epitopes recognised by moABs 130-22 and 145-9 were proved to differ from the CA125 epitope, but to exist on the molecule bearing CA125. Unlike OC125, the majority of 130-22/145-9 activity was associated with a much lower molecular mass (less than 200 kDa), indicating that a lower molecular mass immunoreactive determination may be a unique CA130 antigenic determinant within CA125 molecule. Clinical data demonstrate that, (1) elevated levels of CA130 determinant were found in the sera of 91.3% of women with epithelial ovarian cancer, (2) falling or rising levels of CA130 correlated with regression or progression of ovarian cancer in > 95% of cases, (3) normalisation of serum CA130 levels at response does not imply no microscopic residual disease, but CA130 changes during follow-up support the evaluation of recurrence and can be used as a monitoring marker in an individual patient.     

Swine monoclonal antibodies of high affinity and specificity to carcinoembryonic antigen

To avoid the exclusive use of rodent monoclonal antibodies (MAbs) in patients for the detection of tumors by immunoscintigraphy and for radioimmunotherapy, swine MAbs were produced that are directed against carcinoembryonic antigen (CEA). Spleen cells from 2 pigs immunized with purified colon carcinoma CEA were fused with a nonsecreting mouse myeloma cell line by conventional methods, except that a particularly long immunization protocol and large amounts of spleen and myeloma cells were used. Of 1,200 growing hybrids tested, 20 were found initially to produce antibodies binding to radiolabeled CEA. Seven stable clones producing anti-CEA MAbs for more than 6 months were derived from these hybrids by repeated subcloning. The pig origin of the seven MAbs was demonstrated in a solid-phase CEA enzyme immunoassay where anti-pig immunoglobin (Ig) antibodies coupled to peroxidase gave a positive reaction while anti-mouse Ig antibodies were entirely negative. All swine MAbs were of the IgG isotype. Three anti-CEA MAbs showed no cross-reactivity with granulocytes, while four others gave various degrees of reactivity with different granulocyte glycoproteins. Competitive binding to CEA performed for two purified swine MAbs showed that they recognized two different epitopes. The affinity constants measured for these two MAbs by Scatchard plot on purified CEA were high (1.2 X 10(9) and 1.2 X 10(10) liter/mol). One of the MAbs was tested in vivo for tumor localization by injection, after radiolabeling, in nude mice bearing human colon carcinoma xenograft. High ratios of tumor to normal tissue were obtained with mean values of 10.5 for intact MAbs and of 26.8 for F(ab')2 fragments of the porcine MAb. The results showed that heterofusion with this particular protocol can be used to produce swine MAbs of high affinity and specificity for a well-defined tumor marker. These reagents may have an important clinical utility, particularly in patients who became sensitized to mouse immunoglobulins.     

A low pH enzyme linked immunoassay using two monoclonal antibodies for the serological detection and monitoring of breast cancer

A new, simple and sensitive low pH ELISA method has been developed to measure serum levels of tumour associated antigens detectable by monoclonal antibodies HMFG1 and HMFG2. We examined sera from healthy controls, patients with neoplastic and non-neoplastic conditions of breast, liver and gastrointestinal tract. The majority of patients with metastatic breast cancer had elevated serum antigens (69% HMFG1, 72% HMFG2) compared to healthy controls (6.3% HMFG1, 3.0% HMFG2) or patients with benign breast disease (17% HMFG1, 4% HMFG2). There was no discrimination using these assays between patients with neoplastic and non-neoplastic conditions of liver and gastrointestinal tract. This new method promises to be of value in the assessment and management of patients with breast cancer.     

CEA mRNA 、CEA和CA19-9检测在良、恶性胸水诊断中的应用

目的：研究检测癌胚抗原信使核糖核酸（CEA mRNA）、癌胚抗原（CEA）和糖链蛋白抗原19－9（CA19－9）水平在诊断与鉴别诊断良、恶性胸水中的应用价值。方法：恶性胸水组76人，良性胸水组58例，采集胸水，应用反转录套式聚合酶链反应（RT－NP－PCR）技术检测CEA mRNA，磁分离酶免疫技术（MAIA）检测CEA和CA19－9。结果：CEA mRNA、CEA和CA19－9的阳性率，恶性胸水组分别为78.9%（60／76）、52.6%（40／76）和55.3%（42／76），良笥胸水组分别为8.6%（5／58）、5.2%(3/58)和3.4%（2／58），良、恶性胸水组各 示阳性结果的差异均具有显著性（P＜0.001）。检测CEA mRNA、CEA和CA19－9水平诊断恶性胸水的特异性分别为91.4%、94.8%和96.6%，敏感性分别为78.9%、52.6%和55.3%。三个指标联检的特异性为90.8%、敏感性为82.8%。结论：CEA mRNA、CEA和CA19－9在恶性胸水中有较高的阳性率，其中以CEA mRNA指标的阳性率最高，应用CEA mRNA、CEA和CA19－9指标诊断与鉴别诊断良、恶性胸水具有较高的实用价值，三者联检可以进一步提高敏感性。     

GENERATION AND CHARACTERIZATION OF TWO MONOCLONAL ANTIBODIES AGAINST CYTOKERATINS

Thirty-five hybridoma cell lines against cytoke-ratins, isolated from Hela cells and human cellus respectively, were generated by fusion of immunized spleen cells of BALB/C mice with P3×63-Ag8.653, a mouse myeloma cell line. Two of them (HI and C53) were characterized by indirect immu-nofluorescence, ABC immunostaining and immuno-blotting. The results of immunofluorescence and ABC immunostaining suggested that both monoclonal antibodies were specific for keratin-type intermediate filaments. However, the two monoclonal antibodies showed different specificities in normal tissues and neoplasms as observed on both frozen and deparaf-finized sections. In normal tissues, H1 stained transitional epithelium and all types of simple epithelium except endothelium and mesothelium but did not stain stratified squamous epithelium. In contrast, C35 recognized only stratified squamous epithelium, but failing to react with simple epithelium. Both monoclonal antibodies did not react with nonepithelia cells and tissues.     

Production and characterization of monoclonal antibodies with specificity for human HLA-G isoforms

Production of monoclonal antibodies to HLA-G, a nonpolymorphic antigen of non-classical HLA class I, is of basic and clinical importance. In the present study, monoclonal antibodies were prepared which recognize different membrane bound and soluble isoforms of HLA-G, following immunization of BALB/c mice with a synthetic peptide. Use of this peptide (23 residues), which is present in the alpha1 domain of HLA-G, was previously demonstrated to provide antibodies useful for recognition of HLA-G isoforms in human placenta. Antibody-producing hybridomas were screened by an indirect one-step ELISA method. A clone designated 5E6H7, secreted antibodies useful in immunostaining studies involving both surface HLA-G of placental tissues and soluble forms of this antigen in human sera. In addition, unreactivity of this antibody with human lymphocytes and sections of normal human skin was observed by immunofluorescence microscopy, thus demonstrating its specificity for HLA-G.     

消化道恶性肿瘤TSGF、CA19-9、CA242联合检测的临床评价

目的　探讨用恶性肿瘤特异性生长因子 (TSGF)、糖抗原 19 9(CA19 9)与糖抗原 2 42 (CA2 42 )联合检测对消化道恶性肿瘤的临床评价。方法　采用酶免疫测定 140例消化道肿瘤及 30例正常人血清。结果　以单一指标阳性作为诊断标准CA2 42、CA19 9、TSGF对消化道恶性肿瘤的灵敏度分别为 42 9%、5 5 7%、80 7% ,准确度分别为 5 2 9%、6 2 4%、81 8% ,联合检测结果二项或二项以上阳性作为标准则本组患者的诊断的灵敏度为 84 3% ,准确度为 87 6 %。结论　联合检测血清TSGF、CA19 9与CA2 42对消化道恶性肿瘤的辅助诊断有较高的临床应用价值。     

Production and characterization of two human glioma xenograft-localizing monoclonal antibodies

Multiple fusions following immunization of athymic mice with the extensively characterized human glioma cell line D-54 MG resulted in the selection of several antibodies (Mabs) highly reactive with tumors of neuroectodermal origin and unreactive with normal nervous system tissue. Two Mabs, C12 and D12, which localized specifically to tumors in athymic mouse-human glioma xenograft paired label localization assays, are IgG3 antibodies; both bind readily to staphylococcal protein A in column purification and radioimmunoprecipitation procedures. Both iodinate via the chloramine-T method yielding 125I-immunoreactive product by direct cell surface radioimmunoassay and absorption assay. By indirect cell surface radioimmunoassay, a cultured cell line panel consisting of 17 gliomas, 3 medulloblastomas, 2 neuroblastomas, 2 melanomas, and 2 fetal and 2 adult brain-derived cell lines was examined; the two Mabs were highly similar but distinct in their reactivity profiles. Each was positive with greater than 47% of the gliomas tested (C12, 9 of 17; D12, 8 of 17); and with 1 of 3 medulloblastomas, 1 of 2 melanomas, and cell lines derived from 12- and 16-week-gestation human fetal brain. No reactivity was observed with neuroblastoma or adult brain-derived cell lines or with neutral glycolipids and gangliosides extracted from D-54 MG xenografts or human glioma cell lines. Notable extraneuroectodermal reactivity included that of Mab D12 for splenic trabeculae and the spermatids and Sertoli cells in the testes. Following immunoprecipitation of [3H]leucine labeled cell membrane preparations, Mabs C12 and D12 have consistently yielded unique bands in the Mr 180,000 and Mr 88,000 regions respectively. When used in paired label localization experiments in s.c. D-54 MG xenograft-bearing athymic mice, Mabs C12 and D12 demonstrate similar localization patterns, attaining peak localization indices at day 3 (D12) or 4 (C12); the maximum percentage of injected Mab bound to tumor ranged from 5% (D12) to 8% (C12). The peak tumor/normal brain localization ratios (167-181) attained by these Mabs at days 1-2 followed by their rapid clearance suggest that these Mabs are potentially useful imaging and therapeutic agents for further investigation.     

Serum determination of CA 19-9 in patients with digestive cancers and its diagnostic evaluation

The precision of CA 19-9 RIA kit was evaluated by recovery, reproducibility and dilution test with very satisfactory results. The CA 19-9 value in sera from 52 healthy individuals and from 224 patients with gastric intestinal cancer and other benign disease, showed an increased positive rate in several cases of gastric intestinal cancer. For example, the positive rate in pancreatic cancer, bile duct cancer, colo-rectal cancer, gastric cancer, esophagus cancer, primary biliary cirrhosis diabetes mellitus, liver cirrhosis and chronic hepatitis was 60%, 75%, 55.6%, 45.6%, 20%, 28.6%, 22.7%, 13.7% and 1.7% respectively. By contrast, values from patients with acute hepatitis, fulminant hepatitis, fatty liver, gastric duodenal ulcer, pancreatitis, and primary liver cancer were within the normal range. In this study, CA 19-9 RIA were found to be significant as an adjunct in the management of patients with gastrointestinal cancer, especially pancreatic cancer, and bile duct cancer.