cefoxitin and Ovarian-Neoplasms

We have synthesized various formulations that have potential for active specific immunotherapy (ASI) of human cancers. Sialyl-Tn (STn) is a potentially important target structure for ASI because its expression on mucins is a strong, independent predictor of poor prognosis, suggesting that it may have functional significance in the metastatic process. In this first pilot study of synthetic sialyl-Tn hapten conjugated to keyhole limpet hemocyanin (STn-KLH), with Detox adjuvant, toxicity and humoral immunogenicity were assessed in 12 patients with metastatic breast cancer. Toxicity was minimal, restricted to local cutaneous reactions (apart from transient nausea and vomiting following single low-dose cyclophosphamide treatment). Using STn-conjugated human serum albumin in a solid-phase enzyme-linked immunosorbent assay, it was shown that all patients developed IgM and IgG specific for the synthetic STn hapten. Following immunization, most patients were shown to develop increased titres of complement-mediated cytotoxic antibodies, partially inhibited by synthetic STn hapten, but not by the related TF hapten. We also detected IgM and IgG antibodies reactive with natural STn determinants expressed on ovine submaxillary mucin, the STn specificity of this reactivity being confirmed by hapten inhibition. Evaluation of clinical efficacy in a small pilot study is difficult. Five patients are alive 12 or more months after entry, and another 4 patients are alive 6 or more months after entry into the study. All 3 patients with known widespread bulky disease progressed despite ASI, 2 having died from widespread cancer. Two patients had partial responses, each lasting 6 months. While several patients had disease stability for 3-10 months, 1 patient with pulmonary metastases remains stable 15 months after entry into the program.

Topics: Adjuvants, Immunologic; Antibodies, Neoplasm; Antibody Specificity; Antigens, Tumor-Associated, Carbohydrate; Breast Neoplasms; Carcinoma; Complement System Proteins; Cytotoxicity, Immunologic; Female; Glycoconjugates; Haptens; Humans; Immunization; Ovarian Neoplasms; Tumor Cells, Cultured

The Sialyl-Thomsen-nouveau antigen (STn) is abundantly produced on many types of human epithelial cancers including epithelial ovarian cancer (EOC). We previously developed an EOC-specific lectin sandwich immunoassay (CA125. An analytically sensitive CA125. Our findings suggest that Eu

Topics: Antibodies, Monoclonal; Antigens, Tumor-Associated, Carbohydrate; Biomarkers, Tumor; CA-125 Antigen; Carcinoma, Ovarian Epithelial; Case-Control Studies; Cross Reactions; Diagnosis, Differential; Endometriosis; Europium; Female; Humans; Immunoassay; Metal Nanoparticles; N-Acetylneuraminic Acid; Neoplasm Grading; Ovarian Neoplasms; ROC Curve

A high expression of O-glycosylated proteins is one of the prominent characteristics of ovarian carcinoma cells associated with cell migration, which would be attributed to the upregulated expression of glycosyltransferases. Therefore, elucidating glycosyltransferases and their substrates may improve our understanding of their roles in tumor metastasis. In the present study, we reported that knockdown of polypeptide N-acetylgalactosaminyltransferase 14 (GALNT14) by small interfering RNA significantly suppressed the cell migration and altered cellular morphology. Immunoprecipitation and western blot analyses indicated that GALNT14 contributed to the glycosylation of transmembrane mucin 13 (MUC13), which was significantly higher in ovarian cancer cells compared with the normal/benign ovary tissues. Furthermore, interleukin-8 (IL-8), which could regulate the migration ability of epithelial ovarian cancer (EOC) cells, had no remarkable effect on the expression of GALNT14 and the tumor-associated carbohydrate epitope Tn antigen. In addition, extracellular signal-regulated kinase 1/2 (ERK1/2) inhibitor modulated the expression levels of GALNT14. Our findings provide evidence that GALNT14 may contribute to ovarian carcinogenesis through aberrant glycosylation of MUC13, but not through the IL-8 pathway. These data provide novel insights into understanding the function of MUC13 on neoplasm metastasis and may aid in the development of new anticancer drugs for EOC.

Topics: Antigens, Tumor-Associated, Carbohydrate; Carcinogenesis; Carcinoma, Ovarian Epithelial; Cell Line, Tumor; Cell Movement; Female; Glycosylation; Humans; Interleukin-8; MAP Kinase Signaling System; Mucins; N-Acetylgalactosaminyltransferases; Neoplasm Metastasis; Neoplasms, Glandular and Epithelial; Ovarian Neoplasms; Polypeptide N-acetylgalactosaminyltransferase

Antibody-dependent cell cytotoxicity (ADCC) plays a critical role in monoclonal antibody (mAb)-mediated cancer therapy. ADCC, however, has not been directly shown in vivo but inferred from the requirement for IgG Fc receptors (FcγR) in tumor rejection in mice. Here, we investigated the mechanism of action of a Tn antigen-specific chimeric mAb (Chi-Tn), which binds selectively to a wide variety of carcinomas, but not to normal tissues, in both humans and mice. Chi-Tn mAb showed no direct toxicity against carcinomas cell lines in vitro but induced the rejection of a murine breast tumor in 80% to 100% of immunocompetent mice, when associated with cyclophosphamide. Tumor rejection was abolished in Fc receptors-associated γ chain (FcR-γ)-deficient mice, suggesting a role for ADCC. Indeed, tumor cells formed stable conjugates in vivo with FcR-γ chain-expressing macrophages and neutrophils in Chi-Tn mAb-treated but not in control mAb-treated mice. The contact zone between tumor cells and ADCC effectors accumulated actin, FcγR and phospho-tyrosines. The in vivo formed ADCC synapses were organized in multifocal supra-molecular activation clusters. These results show that in vivo ADCC mediated by macrophages and neutrophils during tumor rejection by Chi-Tn mAb involves a novel type of multifocal immune synapse between effectors of innate immunity and tumor cells.

Topics: Animals; Antibodies, Monoclonal; Antibodies, Monoclonal, Humanized; Antibodies, Neoplasm; Antibody Specificity; Antibody-Dependent Cell Cytotoxicity; Antigens, Tumor-Associated, Carbohydrate; Antineoplastic Agents, Alkylating; Breast Neoplasms; Cell Line, Tumor; Combined Modality Therapy; Cyclophosphamide; Cystadenocarcinoma, Serous; Female; Humans; Immunization, Passive; Immunological Synapses; Macrophages; Mammary Neoplasms, Experimental; Mice; Mice, Inbred BALB C; Neutrophils; Ovarian Neoplasms; Receptors, IgG; Trastuzumab; Tumor Microenvironment

The object of this study was the investigation of carbohydrate antigen expression in malignant epithelial cells and benign mesothelial cells in serous effusions from patients diagnosed with epithelial ovarian carcinomas. In addition, to compare antigen expression in carcinoma cells in effusions with those of corresponding primary tumors and metastatic lesions. Sections from 63 malignant effusions from ovarian carcinoma patients and 15 reactive effusions were immunohistochemically stained, using 5 monoclonal antibodies for Lewis(y), Sialyl Lewis(x), Tn, and Sialyl Tn antigens. Tissue sections (n = 97) from corresponding primary ovarian carcinomas and metastatic lesions, as well as from 12 malignant mesotheliomas, were additionally stained using the above panel. Staining for the 4 antigens was seen in carcinoma cells in serous effusions in the majority of cases (range = 71% to 85%). In contrast, immunoreactivity was detected in mesothelial cells in only 6% to 23% of the specimens studied (P < .001 for all 5 markers). With the exception of B3 antibody against Lewis(y) antigen, malignant mesotheliomas stained negative, infrequently showing focal immunoreactivity. An up-regulation of Tn and Sialyl Tn expression was detected in carcinoma cells in effusions when compared with both primary tumors (P < .003 and P < .007, respectively) and metastatic lesions (P < .034 and .041, respectively). Cancer-associated carbohydrate antigens can thus be used as an adjunct in the differentiation between malignant epithelial and reactive mesothelial cells. Ovarian carcinoma cells in effusions show up-regulation of Tn and Sialyl Tn, possibly representing a transient phenotypic alteration facilitating metastasis.

Topics: Antigens, Tumor-Associated, Carbohydrate; Ascitic Fluid; Biomarkers, Tumor; Carcinoma; Exudates and Transudates; Female; Humans; Immunoenzyme Techniques; Lewis Blood Group Antigens; Mesothelioma; Neoplasms, Mesothelial; Ovarian Neoplasms; Pleural Effusion, Malignant; Up-Regulation

Human polyclonal, monospecific anti-T and -Tn antibodies were found to be reactive in ELISA tests with human ovarian (IGROV-1, OVCAR-3 and SKOV-3), breast (SKBr-3 and T47D)- and oral (KB)-carcinoma cell lines, but less so or non-reactive with normal epithelia and fibroblasts. The direct binding radioimmunoassay, using 125I-labeled human antibodies, to the IGROV-1 cancer cells was inhibited by homologous unlabeled antibodies of the same concentration, but not by the respective immunodominant haptenic monosaccharides (Gal for T and GalNAc for Tn). Rodent ascitic monoclonal anti-T (Ca3114 and Ca3741) and anti-Tn (Ca3250, Ca3268 and Ca3638) antibodies were also reactive with the ovarian- and breast-cancer cells, as measured by FACS and ELISA tests, but to a lower extent than the polyclonal human antibodies. Both the monoclonal anti-T (Ca3741) and anti-Tn (Ca3250 and Ca3638) antibody-binding reactivities were significantly inhibited by the haptenic free monosaccharides. Addition of the above MAbs to IGROV-1 ovarian-cancer or T47D breast-cancer cells cultured in vitro resulted in significant cytological change and inhibition of the viability of the tumor cells, but not of normal epithelial breast cells. This effect on viability was shown to be complement-independent, yet it was profoundly influenced by the concentration of the serum added to the assay medium. In vivo biodistribution of the anti-T (Ca3114) and anti-Tn (Ca3638) MAbs administered i.p. to athymic IGROV-1 tumor-bearing CD1 female nude mice revealed higher 125I-labeled antibody accumulation in the tumor xenografts and in their lung tissues, as compared with other organs of the same mice tested. The above results thus suggest the feasibility of utilizing these antibodies in immunotherapy and drug targeting.

Topics: Animals; Antibodies, Monoclonal; Antibody Specificity; Antigens, Tumor-Associated, Carbohydrate; Antigens, Viral, Tumor; Breast Neoplasms; Carcinoma; CCAAT-Enhancer-Binding Proteins; Cell Survival; Coculture Techniques; DNA-Binding Proteins; Dose-Response Relationship, Immunologic; Enzyme-Linked Immunosorbent Assay; Female; Humans; Mice; Mice, Nude; Neoplasm Transplantation; Nuclear Proteins; Ovarian Neoplasms; Tumor Cells, Cultured

Altered glycosylation of mucins leading to the expression of T, Tn, and sialyl-Tn antigens has been shown in ovarian carcinoma, but its relationship with prognosis is still unclear. We investigated immunohistochemically the expression of these antigens in 38 (17 serous and 21 mucinous) ovarian carcinomas to assess their potential prognostic value as compared with stage of disease, histopathology of tumors, and survival time of patients. Eight benign ovarian tumors (four serous and four mucinous), and four normal ovarian tissues also were studied. Of the 38 carcinomas, 25 (66%) expressed T, 27 (71%) expressed Tn, and 33 (87%) expressed sialyl-Tn antigens. Most cases (83%) expressed two or all of the three types of antigens simultaneously. Normal ovarian epithelia showed no staining for these antigens, and benign ovarian tumors were either negative or occasionally expressed weak staining in less than 25% of epithelial cell areas. Statistical analyses showed strong associations between Tn and sialyl-Tn antigen expressions and disease stage as well as histological grade. In 19 ovarian carcinoma patients with available survival data, the overall survival times of patients with high Tn or sialyl-Tn antigen expression were significantly worse than those of the patients with negative and low expression (P < .05 and P < .01). In multivariate stepwise regression analysis, disease stage (P = .000) and Tn antigen expression (P = .02) were found to be significant independent parameters associated with the overall survival time. These findings suggest that, with exception of T antigen expression, the expression of Tn and sialyl-Tn antigens in ovarian carcinomas may provide additional prognostic information on patient outcome.

Topics: Antigens, Tumor-Associated, Carbohydrate; Biomarkers, Tumor; Carcinoma; Female; Humans; Immunohistochemistry; Ovarian Neoplasms; Prognosis; Survival Rate

Carcinoma-associated Tn and sialyl-Tn antigens have been shown to aid in the prediction of tumor aggressiveness. The purpose of this study was to evaluate the prognostic value of Tn and sialyl-Tn antigen expression in human ovarian carcinoma.. Formalin-fixed, paraffin-embedded tissue sections from 32 primary ovarian carcinomas, 6 benign and 2 normal ovarian tissues were immunostained using monoclonal antibodies against Tn and sialyl-Tn antigens and a streptavidin-biotin-peroxidase method Immunostainings were assessed semiquantitatively and the measurements were then correlated with the established prognostic factors of ovarian carcinoma. i.e. disease stage and histological grade.. Of the 32 ovarian carcinomas, 22 (69%) expressed Tn and 28 (87.5%) sialyl-Tn antigens. Immunostaining measures for Tn antigen were significantly associated with the clinical stage (p < or = 0.02) and histological grade (p < or = 0.05) of the tumors. The results for sialyl-Tn antigen revealed more significant associations with the clinical stage (p < or = 0.002) and histological grade (p < or = 0.007). The clinical stage and histologic grade of the tumors were also highly correlated with each other. Similarly, combined Tn and sialyl-Tn antigen expression revealed significant correlations with the prognostic factors. Benign and normal ovarian tissues showed no reactive Tn and sialyl-Tn antigen.. The extent of Tn and sialyl-Tn antigen expression in primary ovarian carcinoma may contribute to the prognosis of the disease.

Topics: Antigens, Tumor-Associated, Carbohydrate; Biomarkers, Tumor; Female; Humans; Immunoenzyme Techniques; Ovarian Neoplasms; Prognosis

To determine whether the monoclonal antibody (MoAb) 83D4, previously shown to be highly specific for carcinoma cells, can be used as an immunocytological marker to discriminate between benign and malignant cells in serous effusions; and to test for a correlation between expression of the antigen reacting with MoAb 83D4 on effusion cells and the amount of soluble 83D4 antigen in effusion fluids.. Thirty three pleural and 23 peritoneal effusions from 56 cancer patients with metastatic disease were tested for the presence of Tn associated 83D4 antigen by immunocytochemical staining, and for the presence of soluble antigen in supernatants. The patients had undergone various chemotherapy and radiation therapy protocols.. As a result of the various types of treatment, the cytological characteristics of the cells were often modified and the antigenic epitopes may have been altered. Positive staining for 83D4 MoAb was obtained in 36 (97%) of the 37 malignant effusions, eight (73%) of 11 suspect effusions, and three (38%) of the eight apparently benign effusions (free of malignant cells). In these latter cases, cytological reassessment showed a few suspect cells in two cases. 83D4 soluble antigen was detected in 30 of 37 malignant effusions (81%), five of 11 suspected infusions (46%), and five of eight apparently benign effusions (63%).. Immunocytochemical staining with anti-83D4 antibody is useful for differentiating reactive or atypical mesothelial cells from epithelial cells, especially in breast cancer effusions.

Topics: Adenocarcinoma; Antibodies, Monoclonal; Antigens, Tumor-Associated, Carbohydrate; Ascitic Fluid; Biomarkers, Tumor; Breast Neoplasms; Female; Humans; Ovarian Neoplasms; Pleural Effusion, Malignant; Solubility

Many of the cancer-associated antigens recently have been identified as mucin antigens. However, there are no detailed studies describing the expression of carbohydrates and core proteins of mucin antigens in ovarian tumors. In this study we examined the expression of carbohydrate antigens, which are associated with the earliest steps in mucin glycosylation (Tn and sialosyl-Tn), and the expression of the mucin core protein antigens associated with the MUC1 gene product (mammary-type apomucin) and the MUC2 gene product (intestinal-type apomucin) in 123 ovarian epithelial (mucinous and serous) tumors. In normal ovarian tissues neither Tn, sialosyl-Tn, nor intestinal-MRP antigens (MUC2 gene product) were expressed, except for positive sialosyl-Tn staining of stromal capillaries, while the MUC1 gene product, DF3 antigen, was expressed in the cell apex of the germinal coelomic epithelium when it had plump, slightly elongated, or pseudostratified nuclei. In the benign adenomas Tn and sialosyl-Tn antigens were detected in a small number of mucinous adenomas and rarely in serous adenomas. In contrast, expression of both Tn and sialosyl-Tn antigens was observed in all the adenocarcinomas and in a considerable number of borderline malignancies. DF3 antigen was expressed in many benign serous tumors but not so frequently in benign mucinous tumors; however, it was frequently expressed in the adenocarcinomas and borderline malignancies of both mucinous and serous types. Intestinal-MRP antigen expression increased with the transition of the mucinous tumors from a benign to malignant state, although it was never detected in the serous tumors. Coexpression of DF3 and intestinal-MRP antigens was seen in borderline malignancies and carcinomas of the mucinous tumors. In conclusion, simultaneous expression of Tn and sialosyl-Tn antigens is a highly effective tumor marker in both mucinous and serous tumors of the ovary. Coexpression of DF3 and intestinal-MRP antigens may indicate the malignant potential of ovarian mucinous tumors.

Topics: Adenocarcinoma; Antigens, Neoplasm; Antigens, Tumor-Associated, Carbohydrate; Biomarkers, Tumor; Cystadenoma; Female; Humans; Immunoenzyme Techniques; Mucins; Ovarian Neoplasms

The expression of sialyl-Tn and Tn antigens in various benign, borderline, and malignant ovarian tumors was examined immunohistochemically using newly developed antibodies specific for sialyl-Tn and Tn antigens. Sialyl-Tn antigen was detected in only one benign tumor, a mucinous adenoma that showed faint cytoplasmic staining in a few cells. However, sialyl-Tn was present in 5 of 12 serous borderline tumors, 10 of 19 mucinous borderline tumors, 10 of 13 serous adenocarcinomas, 15 of 16 mucinous adenocarcinomas, 14 of 15 endometrioid adenocarcinomas, and 7 of 7 clear cell carcinomas of the ovary. The antigen expression was observed throughout the cytoplasm of cancer cells and in the apical cytoplasm and luminal contents of some glands. The incidence and intensity of staining for sialyl-Tn antigen were higher in malignant tumors than in borderline tumors, but these results did not correlate with the histologic classification or differentiation. Coexpression of sialyl-Tn antigen and Tn antigen was observed in two serous adenocarcinomas, six mucinous borderline tumors, five mucinous adenocarcinomas, eight endometrioid, and seven clear cell carcinomas. In no case was Tn antigen expressed without concomitant sialyl-Tn antigen expression. Accumulation of sialyl-Tn antigen seems to be an early event of carcinogenesis of the ovary.

Topics: Antigens, Neoplasm; Antigens, Tumor-Associated, Carbohydrate; Biomarkers, Tumor; Female; Humans; Immunohistochemistry; Ovarian Neoplasms; Staining and Labeling