cefoxitin and Hyperplasia

The tumor-associated Tn antigen has been investigated extensively as a biomarker and therapeutic target. Cancer vaccines containing the Tn antigen as a single tumor antigen or as a component of a polyvalent vaccine have progressed into phase I and II clinical trials. One major focus of Tn-based vaccines is the treatment of prostate cancer patients. Although expression of the antigen on prostate tumors is a critical prerequisite, previous reports investigating Tn expression in prostate tumors have produced conflicting results. Using a combination of immunohistochemistry and carbohydrate microarray profiling, we show that only 4% to 26% of prostate tumors express the Tn antigen. Based on our results, the majority of prostate cancer patients do not express the appropriate antigen. Therefore, efforts to preselect the subset of prostate cancer patients with Tn-positive tumors or apply Tn vaccines to other cancers with higher rates of antigen expression could significantly improve clinical response rates. Because conflicting information on carbohydrate expression is a general problem for the field, the approach described in this article of analyzing antigen expression with multiple antibodies and using carbohydrate microarray profiles to interpret the results will be useful for the development of other carbohydrate-based cancer vaccines and diagnostics.

Topics: Adenocarcinoma; Aged; Animals; Antigens, Tumor-Associated, Carbohydrate; Biomarkers, Tumor; Cancer Vaccines; Carbohydrates; Carcinoma, Transitional Cell; Clinical Trials as Topic; Humans; Hyperplasia; Male; Prostatic Neoplasms; Rabbits

VVA-B4 lectin was used to investigate the differences in Tn antigen expression in tissues of different types of human breast cancer (benign lesions, carcinoma in situ, invasive carcinoma) and in normal tissues neighboring lobular carcinoma. Locations in which Tn antigen was expressed were identified using the avidin-biotin-peroxidase labeling system. Tissues collected during cosmetic procedures and classified as normal were completely negative, except for one case. Benign proliferative changes including fibroadenoma, apocrine and cylindrical metaplasia showed a very weak positive reaction, although strongly positive cells were also observed. The reaction in non-invasive cases of atypical hyperplasia was diversified depending on site. Intralobular hyperplasia was characterized by a particularly high percentage of labeled cells. A majority (up to 80%) of ductal and lobular carcinoma in situ showed very strong or moderate staining. In invasive cancers, there were conspicuous differences between stage of cancer development and tendency towards a decrease in intensely labeled cell count in the most advanced stages. In normal tissues in the direct neighborhood of carcinoma in situ, the cytoplasm of 40% of cells was strongly labeled. However, the findings for normal tissues in the close vicinity of invasive cancer were the most surprising, since there was either no or only very weak positive reaction. It can be concluded that glycosylation modifications during carcinogenesis, as demonstrated by the presence of Tn epitope, develop very early, before any destructive changes in proliferation/apoptosis or cell differentiation become discernible.

Topics: Antigens, Tumor-Associated, Carbohydrate; Biomarkers, Tumor; Breast Neoplasms; Carcinoma in Situ; Carcinoma, Ductal, Breast; Carcinoma, Intraductal, Noninfiltrating; Cell Transformation, Neoplastic; Female; Fibroadenoma; Humans; Hyperplasia; Immunohistochemistry; Lectins; Precancerous Conditions

Expressions of the carcinoembryonic Tn antigen studied with VVA-B4 and GSI-A4 lectins with the monoclonal antibody 83D4 and of N-acetyllactosamine residues with ECA and LSL lectins, were examined in 54 malignant or benign human breast tumors. Positive membrane labelling with lectins and 83D4 MAb occured in benign cases indicating that modification of glycoconjugates may precede the cytologic anomalies. In fibroadenoma, fibrocystic dystrophy, ductal hyperplasia and grade I invasive ductal carcinomas, the binding sites for all lectins and 83D4 MAb were essentially on the cell membrane with labelling of both apical and basolateral compartments. In grade II and III, the labelling involved the cytoplasm, and cell heterogeneity appeared. The disappearance of reactivity observed for a large proportion of cells at grade III may be due either to the loss of glycosyl-transferase, or to the lack of synthesis of the protein back-bone. Invasive lobular carcinomas showed labelling both on apical membrane and the outermost part of the cytoplasm with a distinct cell polarity. Lectin receptors are present at the surface of metastatic cells, possibly related to their involvement in adhesion.

Topics: Amino Sugars; Antibodies, Monoclonal; Antigens, Tumor-Associated, Carbohydrate; Breast; Breast Neoplasms; Carcinoma, Ductal, Breast; Carcinoma, Lobular; Female; Fibroadenoma; Glycosyltransferases; Humans; Hyperplasia; Immunohistochemistry; Lectins

Expression of carcinoembryonic Tn antigen studied with VVA-B4 and GSI-A4 lectins with the monoclonal antibody 83D4 and of T antigen with LDL and PNA lectins with the monoclonal antibody ZCMO4, were examined in 54 malignant or benign human breast tumors and for MCF-7, T47D and MCF-10A cell lines of human breast tumors origin. For breast tissues, positive membrane labelling with D-GalNAc alpha-O-ser/thr (Tn-antigen) specific lectins and 83D4 MAb occurred in benign cases indicating that modification of glycoconjugates may precede the cytologic anomalies. In fibroadenoma, fibrocystic dystrophy, ductal hyperplasia and grade I invasive ductal carcinomas, the binding sites for lectins and 83D4 MAb were essentially on the cell membrane with labelling of both apical and basolateral compartments. In grade II and III, the labelling involved the cytoplasma, and cell heterogeneity appeared. The disappearance of reactivity observed for a large proportion of cells at grade III may be due either to the loss of glycosyltransferase, or to the lack of synthesis of the protein back-bone. Invasive lobular carcinomas showed labelling both on apical membrane and the outermost part of the cytoplasm with a distinct cell polarity. Lectin receptors are present at the surface of metastatic cells, possibly related to their involvement in adhesion. In all cases, T or sialosyl-T antigens are present at the surface of tumors cells. All cell lines from breast tumors cultured in vitro were labelled with lectins and monoclonal antibodies. The simultaneous presence of Tn and T antigens on the cells, indicates that the expression of Tn antigen is due to a partial but non total deficiency in the beta-1- > 3 galactosyltransferase involved in T-antigen synthesis.

Topics: Adult; Aged; Aged, 80 and over; Antibodies, Monoclonal; Antigens, Tumor-Associated, Carbohydrate; Antigens, Viral, Tumor; Breast; Breast Neoplasms; Carcinoma, Intraductal, Noninfiltrating; Female; Fibroadenoma; Fibrocystic Breast Disease; Glycoconjugates; Humans; Hyperplasia; Lectins; Middle Aged; Neuraminidase; Tumor Cells, Cultured

In a previous report we suggested that T antigen appeared to be associated with gastric carcinoma. To verify this hypothesis and characterize the pattern of expression of simple-mucin type carbohydrate antigens (Tn,sialyl-Tn and T before and after neuraminidase) in normal gastric mucosa and precursor lesions of gastric carcinoma, we studied the mucosa adjacent to 100 cases of gastric carcinoma, gastric biopsies of 60 dyspeptic patients, eight adenomatous polyps and eight hyperplastic polyps. The expression of the antigens was more related to the cell type and underlying lesions than to the coexistence of carcinoma. The most distinctive findings concerned intestinal metaplasia, dysplasia and hyperplastic lesions. In intestinal metaplasia, Tn was found mostly in columnar cells and sialyl-Tn in goblet cells. T was more prevalent in incomplete intestinal metaplasia than in complete. A high prevalence of sialyl-Tn expression and cell membrane immunoreactivity for T antigen, similar to those previously found in gastric carcinomas, were observed in three adenomatous polyps, one hyperplastic polyp, five cases of adenomatous dysplasia in the neighbourhood of intestinal carcinomas and four cases of marked foveolar hyperplasia, three of which were from the mucosa adjacent to diffuse carcinomas. We conclude that adenomatous and hyperplastic lesions share with gastric carcinomas features of aberrant glycosylation, namely the cell membrane expression of T antigen.

Topics: Adenocarcinoma; Adenomatous Polyps; Antigens, Tumor-Associated, Carbohydrate; Carbohydrate Sequence; Dyspepsia; Gastric Mucosa; Gastritis; Gastritis, Atrophic; Glycosylation; Humans; Hyperplasia; Immunoenzyme Techniques; Metaplasia; Molecular Sequence Data; Polyps; Precancerous Conditions; Stomach Neoplasms

Atrophy of villi, with increases in crypt depth and Paneth cell number and size, occurs in chronically isolated (Thirty-Vella) ileal loops in rabbits. These loops are known to be heavily colonized with aerobic bacteria. To study the possible effect of the bacterial overgrowth, 2 experiments were performed. In the first study, two isolated ileal loops were created in each of 14 rabbits. The antibiotic loop was flushed with nonabsorbable antibiotics (neomycin, bacitracin, and gentamycin), whereas the control loop was flushed with saline. The antibiotic solution achieved a reduction in bacterial growth as compared to the loops flushed with saline. In the second study, a single ileal loop was created in each of 20 rabbits. Loops of 11 animals were flushed with an absorbable antibiotic (cefoxitin) and gentamycin, whereas those in 9 other rabbits were flushed with saline. This antibiotic combination achieved an essentially sterile loop. In both experiments, the Paneth cell population and crypt depth were less in antibiotic loops as compared to saline loops, whereas the degree of villus atrophy was nearly equal. These studies suggest a link between the overgrowth of bacteria seen in these isolated loops and the morphologic changes in the crypts.

Topics: Animals; Anti-Bacterial Agents; Bacitracin; Cefoxitin; Escherichia coli; Gentamicins; Hyperplasia; Ileum; Intestinal Mucosa; Neomycin; Pseudomonas aeruginosa; Rabbits