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L K Trejdosiewicz

Publications and source records attributed to L K Trejdosiewicz.

106 records · Page 6Linked to original sources
PubMed1984

Identification of cells from fetal bladder epithelium in human amniotic fluid.

Cultured human amniotic fluid cells consist of five different types of cytokeratin-positive epithelial cells, E-1 to E-5, differing by their size, growth morphology, and cytokeratin pattern, according to our earlier investigations. Using anticytokeratin antibodies in indirect immunofluorescence (IIF) microscopy, we show in this study that cultured urine cells contain four of the cell types found in amniotic fluid. In addition, we used two urothelium-specific antibodies, anti-UMA and anti-Las-86, in combination with cytokeratin antibodies to distinguish urothelium-derived cells in amniotic fluid and urine cell cultures. Two of the epithelial cell types were found to express urothelial antigens and thus to originate from the transitional bladder epithelium. These cells were found in 26 of the 33 amniotic fluid cell cultures and in nine of the ten urine cell cultures.

Amniotic Fluid↗
PubMed1984

Species cross-reactive membrane-associated urothelial differentiation antigen.

An antigen, termed "urothelium membrane antigen" (UMA), confined to urothelium and most abundantly associated with the asymmetric unit membrane of the terminally differentiated luminal cells was identified with a monospecific (absorbed or affinity purified) rabbit antiserum obtained by immunization with membranes from normal bovine urothelium. A cross-species immunofluorescence-positive reaction was observed in normal bladder: Luminal urothelial membrane reacted most intensely. Underlying normal urothelial cells also showed a weaker cytoplasmic reaction. Specific luminal membrane labeling was confirmed by transmission and scanning electron microscopy. Specificity of the anti-UMA differed from the general epithelial reactivity obtained with antibodies to bovine urothelium cytosol. Anti-UMA did not cross-react with keratin, although there was some evidence of a filamentous localization in the cytoplasm of permeabilized cells. UMA was variably expressed by urothelial carcinoma-derived cell lines to a degree apparently related to the degree of cell differentiation, showing the highest positivity on the well-differentiated RT4 and RT112 lines and only a very weak reaction with the anaplastic MGHU-1 (EJ) and T24 lines. On immunoblots, anti-UMA reacted with a peptide of approximately 54,000 daltons.

Animals↗
PubMed1982

Disruption of the keratin filament network during epithelial cell division.

The behaviour of keratin filaments during cell division was examined in a wide range of epithelial lines from several species. Almost half of them show keratin disruption as described previously: by immunofluorescence, filaments are replaced during mitosis by a 'speckled' pattern of discrete cytoplasmic dots. In the electron microscope these ' speckles ' are seen as granules around the cell periphery, just below the actin cortical mesh, with no detectable 10 nm filament structure inside them and no keratin filament bundles in the rest of the cytoplasm. A time course of the filament reorganization was constructed from double immunofluorescence data; filaments are disrupted in prophase, and the filament network is intact again by cytokinesis. The phenomenon is restricted to cells rich in keratin filaments, such as keratinocytes; it is unrelated to the co-existence of vimentin in many of these cells, and vimentin is generally maintained as filaments while the keratin is restructured. Some resistance to the effect may be conferred by an extended cycle time. Filament reorganization takes place within minutes, so that a reversible mechanism seems more likely than one involving de novo protein synthesis, at this metabolically quiet stage of the cell cycle.

Animals↗
PubMed1982

Urothelium-specific antibody and lectin surface mapping of bladder urothelium.

Coupled ligand-colloidal gold complexes were found to provide a convenient approach for the localization of scanning electron microscopy of cell surface membrane antigens and lectin-binding sites on bladder urothelium and for the immunocytochemical identification of urothelial cell populations at different stages of differentiation. The ligands used to prove the membrane were a urothelium-specific rabbit antibody raised to a urothelial membrane-associated antigen (UMA), and two lectins: Concanavalin A (Con A) and peanut agglutinin (PNA). A complex luminal surface distribution pattern was demonstrated by the UMA antigen related to the stage of urothelial cell maturation and differentiation. UMA could be detected on the surface of immature and early differentiating intermediate cells, but was absent from the late differentiation stage, becoming re-expressed as the cells matured and was found in greatest abundance on the terminally differentiated superficial cells. It was absent on cells in benign hyperplasia of the urothelium. Cellular and regional differences in lectin binding to the urothelial cell surface was suggested with Con A receptors localized uniformly over the superficial cells, and PNA receptors confined to linear arrays or occasional clusters over the apical surface but evenly dispersed over the lateral surface of these cells.

Animals↗
PubMed1982

Immunohistochemical localisation of keratin and luminal epithelial antigen in myoepithelial and luminal epithelial cells of human mammary and salivary gland tumours.

Rabbit antisera to human 40-63 000 MW epidermal keratin, one batch with restricted distribution of reactivity from an initial (aK1) and one with "broad spectrum" distribution of reactivity from a late bleeding (aK), and to "luminal epithelial antigen" (aLEA) were applied to formalin fixed paraffin embedded sections of human normal and neoplastic mammary and salivary glands using an indirect immunoperoxidase method. aK1 reacted with myoepithelial cells, aLEA with luminal epithelial cells and aK with both cell types in normal mammary and salivary gland. In breast carcinomas the majority of intraluminal and infiltrating carcinoma cells reacted with aLEA but not with aK1 which reacted only with surrounding myoepithelial cells. aK reacted with both myoepithelial cells and with intraluminal and infiltrating tumour cells. In the salivary gland adenomas the majority of cells reacted with aK, and those cells arranged in a tubular fashion reacted with aLEA.

Adenoma↗
PubMed1982

Immunoelectron microscopic localisation of keratin and luminal epithelial antigens in normal and neoplastic urothelium.

Immunoelectron microscope cytochemistry was carried out on 2% paraformaldehyde fixed, 50 mu sections of normal urothelium and bladder carcinoma cells in culture using antisera raised in rabbits to human 40-63 000 MW epidermal "broad spectrum" keratin and calf urothelial "luminal epithelial antigen" (aLEA) Both the unconjugated and indirect immunoperoxidase-DAB techniques were used before routine embedding. The localisation of both keratin and luminal epithelial antigen (LEA) was similar in normal and neoplastic cells and reaction product was associated not only with tonofilaments but also lining membrane vesicles and on fine filaments in the cytoplasmic ground substance.

Animals↗
PubMed1982

Immunohistochemical demonstration of epithelial and urothelial antigens at the light- and electron microscope levels.

Antiserum raised in rabbits against the cytosol fraction from calf bladder epithelium was absorbed with sera, human erythrocytes and non-epithelial tissues until it became specific for a number of epithelia including urothelium, as demonstrated by immunofluorescence and immunoperoxidase techniques. Electron microscope immunocytochemistry showed that, in the urothelium, the majority of reaction product was present in the superficial cells in association with the plasma membrane, cytoplasmic membrane vesicles and cytoskeleton. After a second series of absorptions with epithelial organs other than bladder, the antiserum was rendered urothelium-specific and the strongest reaction was seen in superficial layer cells. Epithelium and urothelium specific antigens were species cross-reactive and could also be demonstrated in foetal and malignant urothelium. In malignantly transformed mouse bladder epithelium cell lines, the epithelial antigens were associated with the inner surface of the plasma membrane and underlying cytoskeletal elements.

Animals↗
PubMed1981

Cytochemical markers of bladder carcinogenesis.

Enzyme cytochemical and immunocytochemical techniques at the light and electron microscope levels were used to study the distribution of potential markers of chemical transformation in rodent bladders. In rat tumours induced by in vivo treatment with methylnitrosourea, alkaline phosphatase localization was normal on the external surface of the plasma membranes of some cells but abnormal in others where reaction product was seen only on intracellular membranes. 5'-Nucleotidase localization was abnormal in all cells, being seen on endoplasmic reticulum and nuclear membranes only, while in normal bladders only ectoenzyme localization was seen. Heterogeneity of alkaline phosphatase amd 5'-nucleotidase localization was seen on the plasma membranes of these tumours after 15 days in organ culture. Some cells produced enzyme and others did not; in other cells only parts of the membrane reacted heavily, while other regions were negative. In transformed cell cultures and tumours of mouse bladder derived by in vitro treatment of explants with dimethylbenz (a) anthracene, a bimodal pattern of alkaline phosphatase localization was seen. Cells had either normal ectoenzyme reaction product or abnormal intracellular membrane reaction product. 5'-Nucleotidase and ADPase were lost after transformation while cAMP-phosphodiesterase was retained as an ectoenzyme. Mg.ATPase and a cAMP-independent, calcium-insensitive 'protein phosphatase' were induced in transformed cell cultures. An epithelial antigen was detected in the cytoplasm of both normal and transformed cells associated with reticular cytoplasmic ground substance, plasma membrane vesicles and cytoskeletal elements.

5'-Nucleotidase↗
PubMed1981

Colloidal gold markers and probes for routine application in microscopy.

Colloidal gold can be used as an electron-dense cytochemical probe for direct or indirect labelling techniques. Gold markers can be prepared in a size range of 5-150 nm, and show size-dependent shape characteristics and absorption spectra. Size and shape distribution increases with mean particle diameter, with appreciable overlapping between populations of different mean size. Using radio isotope-binding assays, spectrophotometric analysis and an innovative rapid microtitration technique, the effect of pH, ionic strength and protein concentration on gold-protein interaction has been studied. Efficient adsorption of protein to gold occurs at, or near, the pI of the protein. The amount of protein needed to effect stabilization is both a function of pH and of ionic strength, but does not reflect the amount of protein binding for all proteins. There is evidence for multilamellar adsorption of proteins to gold, which is discussed in context of the bioactivity, and stability of the probe. A working protocol for the routine reproducible manufacture of protein-gold probes is given, making use of the microtitration assay.

Colloids↗
PubMed1981

Cell surface distribution of fibronectin in cultures of fibroblasts and bladder derived epithelium: SEM-immunogold localization compared to immunoperoxidase and immunofluorescence..

Expression of cell surface fibronectin in cultures of untransformed fibroblasts is well documented, but little is known of its presence and distribution in cultured epithelial cells. Using species monospecific anti-fibronectin antibodies, the distribution of fibronectin in untransformed fibroblasts and in normal and neoplastic bladder epithelial cells was characterized by indirect labelling experiments using immunogold scanning electron microscopy (SEM). The surface matrix of fibronectin expressed in rodent and human fibroblast cell lines was demonstrated with ease by SEM of gold-tagged second antibodies. However, no fibronectin could be detected on any of the mouse and human bladder epithelium-derived cells studied in single or in mixed epithelial-fibroblast cultures. These SEM-immunogold observations were compared to and confirmed by immunofluorescence and immunoperoxidase microscopy. Immunofluorescence and SEM localization of the fibronectin in the extracellular matrix presented similar distribution patterns but the higher resolution of the SEM provided a more detailed analysis.

Animals↗
PubMed1979

Colloidal gold probes--a further evaluation.

Colloidal gold provides an immuno- and cytochemical marker suitable both for TEM and SEM. Various parameters involved in the preparation and stability of gold markers of various sizes and of probes have been reviewed and modifications in methodology suggested based on a study of factors affecting the performance of the gold marker system. In particular, dependence of protein adsorption on pH conditions correlating with protein pI is further confirmed. Presence of bioactive protein in gold probes is demonstrated and quantitated by radioassay (125I) and agglutination procedures.

Cell Line↗
PubMed1979

Growth enhancing property of human monocytes from normal donors and cancer patients.

Peripheral blood monocytes, but not other leucocytes, from healthy donors, were shown to enhance the DNA synthesis of three cell lines. The effect was very marked on an epithelial (Chang) and lymphoid (CLA-4) cell line derived from normal tissues, and less marked on a carcinoma-derived line (HT-29). The enhancement was demonstrable over a wide range of monocyte:cell line ratios, and some activity was present in supernates from monocytes cultured alone or with a cell line. Furthermore, monocytes from gastric carcinoma patients did not enhance Chang and CLA-4 cells to the same extent, relative to the healthy donors.

Adenocarcinoma↗
PubMedDate not supplied

Human hepatocellular carcinoma: cross-reactive and idiotypic antigens associated with malignant transformation of epithelial cells.

Monoclonal antibodies were isolated following immunization with the HBsAg and alpha-fetoprotein-secreting human hepatoma PLC/PRF/5 ("Alexander") cell line. Three antibodies (K-PLC1, K-PLC2 and K-PLC3) showed evidence of carcinoma-associated reactivity by indirect immunofluorescence. Antibodies K-PLC2 and K-PLC3 reacted only with PLC/PRF/5 cells, but not with any other normal or malignant cell type tested, including the Hep/G2 hepatoma cell line. The reactivity of these antibodies was not removed by absorption with homogenates of either normal liver or a primary hepatocellular carcinoma. These results suggest that K-PLC2 and K-PLC3 identify PLC/PRF/5 idiospecific determinants. Following surface iodination of PLC/PRF/5 cells, immunoprecipitation and analysis on polyacrylamide gels, these specific determinants were found to be of 200,000 and 76,000 daltons, respectively. On the other hand, antibody K-PLC1, although unreactive by immunofluorescence on the majority of normal cell types, including those of lymphoid organs and bone marrow liver cells and most epithelia, was weakly positive on some normal ductal secretory epithelia and was positive on vascular endothelium. However, K-PLC1 reacted strongly with all carcinoma specimens tested, and with most carcinoma-derived cell lines, indicating a large increase in K-PLC1 antigen expression by epithelial cells after malignant transformation. Absorption of K-PLC1 with normal liver homogenate had no affect, but absorption with a hepatocarcinoma homogenate abolished its activity. The K-PLC1 antigen could not be immunoblotted or immunoprecipitated and resolved on polyacrylamide gels; yet it showed the properties of a phospholipid, namely resistance to proteases, extractability with organic solvents and sensitivity to phospholipase C.

Antibodies, Monoclonal↗
PubMedDate not supplied

Urothelial cell transplantation using biodegradable synthetic scaffolds.

Reconstruction of the urinary bladder with bowel to restore storage capacity is associated with significant complications arising from substituting an absorptive, mucus-producing intestinal epithelium for the barrier urothelium of the bladder. To overcome these problems, we are developing a "composite enterocystoplasty" procedure to replace the epithelium of the bowel with autologous in vitro-propagated normal urothelial cells. The aims of this study were to evaluate synthetic biomaterials as delivery vehicles for the cultured urothelial cells and provide support during transfer and cell adherence to the de-epithelialized bowel wall. The surgical compliance of 12 biomaterials was evaluated, along with their ability to support urothelial cell attachment. Transfer of urothelial cells onto biomaterials as single cell suspensions or intact cell sheets was investigated. Seeding of a single cell suspension on to non-woven mesh resulted in poor cell attachment. Seeding onto woven mesh was more efficient, but the most effective transfer method involved producing an intact cell sheet that could be combined with woven, knitted and non-woven biomaterials. Transfer of the cell sheet : mesh complexes onto a de-epithelialized bladder stroma produced a stratified epithelium incorporating the strands of the mesh and expressing urothelial-associated antigens after 48 h in organ culture. Thus, we have developed and evaluated a suitable transfer method for in vitro propagated urothelial cells to be used in "composite enterocystoplasty".

Journal Article↗
PubMedDate not supplied

Intra-tumour C. parvum therapy in gastric carcinoma; a pilot study.

In a preliminary and toxicological study, eight patients with carcinoma of the stomach were treated by Corynebacterium parvum immunotherapy prior to surgical excision. Killed C. parvum suspension was injected at multiple sites directly into the tumour mass with the aid of fibre-optic gastroscope at doses of 1.5 mg, 4 mg or 8 mg. Each patient received two injections of the same dose with an interval of 14 days. The second injections were made 14 to 18 days prior to surgery. No chemotherapy was administered during this treatment. Patients were monitored clinically for signs of toxicity. Immunological parameters were assessed on peripheral blood samples taken immediately before each injection and prior to surgery. Antibody titre to C. parvum was estimated by direct agglutination, while cell-mediated responses to C. parvum and 3M KCl tumour extract and phytohaemagglutinin (PHA) were assessed by lymphocyte transformation. Serum carcinoembryonic antigen (CEA) was measured by radio-immune assay. Tumours removed at surgery were examined histologically for the presence of invading immunocytes. These preliminary data will be discussed with respect to the dose of C. parvum administered.

Carcinoembryonic Antigen↗