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Biomedical subjects

L K Trejdosiewicz

Publications and source records attributed to L K Trejdosiewicz.

At least 91 records · Page 5Linked to original sources
PubMed1986

Phenotypic analysis of cultured melanoma cells. Expression of cytokeratin-type intermediate filaments by the M5 human melanoma cell line.

Expression of intermediate filament (IF) isotypes was studied in six human and two murine melanoma cell lines. With one exception, these lines expressed IFs only of the vimentin type; neurofilament peptides, desmin and GFAP were not detected. However, the M5 human melanoma line also expressed extensive cytokeratin tonofilament arrays, as visualized by immunofluorescence with a panel of eleven monoclonal antibodies and hetero-antisera to cytokeratins; only the keratin 19-specific antibody BA16 did not react. By 2 D gel electrophoresis, five major keratin peptides were detected (keratins 7, 8, 13, 17 and 18), and an additional 57 kD peptide was detected on immunoblots with several antikeratin antibodies. Also observed in M5 cells was focal collapse of tonofilament arrays in mitotic cells. All the melanoma lines tested were positive for S100; M5 and two other cell lines were also positive for the 220-240 kD neuroectoderm-associated cell-surface differentiation antigen defined by monoclonal antibody UJ 127:11. In all the melanoma cell lines, secretion of extracellular matrix proteins (fibronectin, laminin and collagen type IV) was sparse or absent, and all were negative for the epithelial cell markers HMG-1 and HMG-2. Co-expression of keratin and vimentin by a melanoma cell line is discussed in the light of recent controversy concerning expression of cytokeratins by other neoplasms of putative neuroectodermal origins.

Antibodies, Monoclonal↗
PubMed1986

Relationship between fibronectin and lymphoid cells in buccal mucosa, labial salivary glands and palatine tonsil.

The distribution of fibronectin in human buccal mucosa and labial salivary glands and its relationship to lymphoid cells was studied using an immunoperoxidase technique and monoclonal and polyclonal antibodies. In all specimens fibronectin was associated with basement membranes of epithelia, vascular endothelium and perineural sheaths. In histologically normal areas of buccal mucosa fibronectin was distributed as a sparse network in the superficial lamina propria. A more extensive network of fibronectin was found within the lamina propria of mucosal specimens infiltrated with T lymphocytes, whereas fibronectin was absent in areas occupied by B lymphocytes. A similar relationship between lymphocyte type and the presence of a fibronectin network was found in labial glands and palatine tonsil. Fibronectin was not detected within oral, salivary gland, tonsillar crypt or capsular epithelium.

B-Lymphocytes↗
PubMed1986

Systemic and local antibody responses to gastric Campylobacter pyloridis in non-ulcer dyspepsia.

Antibody titres to Campylobacter pyloridis in serum and gastric juice were estimated by an enzyme linked immunosorbent assay (ELISA) to whole organisms obtained from bacterial culture in 39 patients with non-ulcer dyspepsia. Whereas 20 of the 21 patients with chronic gastritis had gastric C pyloridis, 17 patients with no C pyloridis had normal histology in the gastric antrum and body. Significantly raised serum IgG and IgA antibody titres to C pyloridis were found in colonised patients with gastritis. Patients with raised IgG antibody to C pyloridis were also shown to have significantly raised titres to other Campylobacter species, suggesting antigenic cross reactivity. Gastric juice antibodies were also studied and IgA titres to C pyloridis were detected in a proportion of patients with gastritis, together with low levels of IgM, but no IgG.

Antibodies, Bacterial↗
PubMed1986

Tissue culture model of transitional cell carcinoma: characterization of twenty-two human urothelial cell lines.

Twenty-two continuous cell lines derived from normal and neoplastic urothelium, maintained under identical culture conditions, were characterized in terms of isozyme phenotype, tumorigenicity, and xenograft morphology following xenotransplantation to nude mice, cytological appearance, in vitro growth rate, labelling index, and colony-forming efficiency, in parallel with separate studies of in vitro drug sensitivities and monoclonal antibody reactivities. Three groups were identified: (a) distinct lines with differing isozyme patterns, a broad spectrum of growth characteristics, and xenograft morphologies similar to the histopathology of the parent tumors after periods of up to 17 yr following establishment in vitro; (b) cross-contaminated sublines (maintained separately in different laboratories for periods of up to 10 yr), with identical isozyme patterns and similar growth characteristics, but differing markedly in tumorigenicity and xenograft morphology; and (c) lines derived from normal urothelium which were nontumorigenic and had an isozyme pattern usually only encountered in untransformed cells. These data indicate that cell lines representative of human transitional cell carcinomas can be selected on the basis of xenograft morphology and isozyme patterns, and that a panel of lines derived from normal and neoplastic urothelium could provide a model system to study the biology and treatment of this disease.

Carcinoma, Transitional Cell↗
PubMed1985

Growth factor production during multistage transformation of epithelium in vitro. I. Partial purification and characterisation of the factor(s) from a fully transformed epithelial cell line.

Transforming growth factor (TGF)-like activity is characterised from one of a series of salivary epithelial cell lines, CSG 211, chemically transformed in vitro. In this transformation system, we can demonstrate multiple stages in the acquisition of a malignant phenotype by normal diploid ductal epithelial cells from male mouse submandibular gland. The fully transformed, tumorigenic cell TGF-like activity in serum-free supernatants resembles no other well-characterised growth factor and has an apparent molecular weight (Mr) of 14 kd. There is also evidence of a higher Mr activity, which is separable by anion exchange chromatography. We show that the premalignant, nontumorigenic progenitor cells of this line do not produce demonstrable TGF-like activity and that this property is therefore acquired as CSG 211 cells become carcinoma producing.

Animals↗
PubMed1985

Antibodies to normal human colon membranes: preparation, characterization and tissue distribution.

Rabbit antisera were raised to a membrane fraction of normal human colonic epithelium. After absorption, two antisera appeared to show restricted epithelial specificity when tested on routine paraffin wax-embedded histological sections by the indirect immunoperoxidase technique. The reaction was intense on epithelial cells of large and small intestine, and positive on stomach and duodenum, bile ducts in liver, gall bladder, pancreas and salivary gland. A very weak reaction was also seen in the bronchus and lung. There was no reaction with stromal, vascular or muscle components. All other tissues tested were negative, including hepatocytes, ectodermally-derived glandular epithelia, urogenital tissues and lymphoid organs. The antigen was also detected in 21 primary and metastatic large bowel carcinomata. By immunofluorescence, the antisera reacted with the colon adenocarcinoma-derived HT29 cell line and with primary colon epithelium explant cultures, but not with cultured fibroblasts. By immunoblotting of HT29 whole cell lysates, a triplet of polypeptides of approximate molecular weight range 55 000 to 60 000 were detected. This specificity appeared to be unrelated to previously described normal or tumour-associated antigens by criteria of tissue distribution, immunolocalization, molecular weight, and either absorption or radiobinding assays, or both.

Antibodies↗
PubMed1985

Micro-heterogenous expression of peanut agglutinin-binding sites in the extracellular matrix of cultured cells.

Using double-label techniques with fluorochrome-conjugated peanut agglutinin (PNA) and indirect immunofluorescence with rabbit species-specific anti-fibronectin antibodies and a mouse monoclonal anti-fibronectin, the extracellular matrix (ECM) of cultured human and mouse fibroblasts (Hell7 and 3T3K) and human bladder epithelial cells (T24) was studied. The antibodies and PNA co-localized extensively. However, a small but consistent degree of micro-heterogeneity was revealed insofar as both PNA-positive fibronectin-negative fibrils as well as PNA-negative fibronectin-positive fibrils were observed. Fibronectin production by T24 cells (but not fibroblasts) was influenced by the growth medium, but this did not affect the heterogeneity. Trypsin removed most cell-surface fibronectin and all PNA-binding sites, but did not account for the observed phenomenon. Intracellular fibronectin, whether present naturally or induced to accumulate by culture in presence of Monensin, was PNA-negative. These data suggest that PNA-binding sites appear on fibronectin as a consequence of incorporation into the extracellular matrix, and that the resultant heterogeneity of spatial expression of beta-galactose-like residues may offer a mechanism whereby mesenchymal cells could modulate the behaviour of overlying cell-types.

Animals↗
PubMed1985

Monoclonal antibodies to human urothelial cell lines and hybrids: production and characterization.

Eleven independent monoclonal antibodies, the LBS series, were isolated after immunization of mice with RT112 cells, a continuous cell line derived from a transitional cell carcinoma of the human bladder. These antibodies were tested by indirect immunofluorescence on a panel of 28 human cell lines, of which 17 were urothelial carcinoma-derived, 4 of non-urothelial carcinoma origin, 3 fibroblast cell lines, 4 lymphoblastoid lines and 7 murine cell lines. Also tested were 7 somatic cell hybrid clones derived by fusion of human RT112 cells with murine bladder carcinoma MB63T/H cells. None of the LBS antibodies reacted with mesenchyme-derived cells, although all reacted with RT112 cells. On the basis of reactivity with the cell line panel, the antibodies were divided into 3 groups. Group I (LBS-1 and 19) reacted with all human epithelium-derived cell lines. Group II (LBS-2, 8, 15 and 17) reacted only with human urothelium-derived cells, tending to recognise the least anaplastic types. Group III antibodies (LBS-10, 20A, 20B, 21 and 34) were urothelium-specific on the human continuous cell line panel, but additionally reacted with murine urothelial and epithelial cell lines. The 6 human-specific antibodies (Group I and II) were used for preliminary analysis of human gene expression in a series of 7 mouse X human urothelial somatic cell hybrids. Each hybrid reacted with at least 1 LBS antibody, although there were changes in gene expression with time in culture, indicating both loss and unmasking of human genes. These data suggest the LBS-series antibodies recognise different determinants associated with epithelial and urothelial cell differentiation, and thus may be valuable probes in the study of normal differentiation and malignant transformation in human urothelial cells.

Animals↗
PubMed1985

A methodological basis for SEM autoradiography: biosynthesis and radioligand binding.

A method is described for scanning electron microscope (SEM) autoradiography whereby preservation of high resolution cell surface details is retained together with degelatination of the emulsion without gross loss or redistribution of silver grains. This method should provide a convenient medium-sized marker for SEM (using secondary, backscattered electron and X-ray imaging) topographic studies of biosynthesized molecules, and of cell surface receptors and antigens, using indirect or direct labelling procedures with radio-labelled ligands.

Autoradiography↗
PubMed1985

Analysis of the antigenic composition of liver specific lipoprotein using murine monoclonal antibodies.

Eight monoclonal antibodies have been raised to antigenic determinants within the liver specific lipoprotein complex. Five of these determinants were species and the others non-species specific. No liver specific determinants were identified. Liver specific lipoprotein antigens recognised by the eight monoclonal antibodies were located either on the hepatocyte membrane and/or along the sinusoidal lining wall or in the cytoplasm of liver parenchymal cells. All antibodies but one reacted with the cell membrane of viable human hepatocytes. The molecular weights of four liver specific lipoprotein-determinants were determined by immunoprecipitation. These ranged from 22 000 to 164 000 daltons.

Adult↗
PubMed1985

Expression of the common acute lymphoblastic leukaemia antigen (CALLA gp100) in the brush border of normal jejunum and jejunum of patients with coeliac disease.

Expression of the gp100 common acute lymphoblastic leukaemia antigen (CALLA) was studied in the mucosa of the gut by means of indirect immunofluorescence on cryostat tissue sections with a panel of eight monoclonal antibodies to common acute lymphoblastic leukaemia antigen (anti-CALLA antibodies) and two antibodies to non-CALLA leukaemic antigens. Expression of CALLA was absent from normal stomach epithelium, adult and fetal colonic epithelium of normal histology, and colonic epithelium from patients with Crohn's disease or ulcerative colitis. By contrast, all eight anti-CALLA antibodies gave a characteristic reaction in normal adult and fetal small bowel mucosa, with specific localisation to the entire brush border of jejunal epithelium. Whereas seven of these antibodies reacted both with normal jejunal epithelium and with the damaged epithelium of patients with coeliac disease, antibody RFAL-2 reacted strongly only with histologically normal small bowel but more weakly in patients with coeliac disease to a degree related to the amount of histological abnormality. Expression of the moeity like CALLA identified with RFAL-2 was strongest in crypt epithelium and proportionally diminished along the villi according to the amount of histological damage in coeliac disease, being essentially absent in patients with "subtotal villous atrophy."

Adult↗
PubMed1985

Characteristics of four new human cell lines derived from squamous cell carcinomas of the head and neck.

Four human cell lines were established from biopsy specimens of squamous cell carcinomas of the larynx (TR131 and TR138), tongue (TR126), and buccal mucosa that had infiltrated a lymph node (TR146). All 4 lines readily formed colonies on a plastic substratum, but they were virtually incapable of forming colonies in an anchorage-independent semisolid support system of soft agar (cloning efficiencies, less than 0.02%). The proliferation of this group of tumor-derived cell lines, therefore, appeared to be highly anchorage dependent. Keratin filaments could be visualized in each line by indirect immunofluorescence with the use of polyclonal or monoclonal antibodies to keratins; staining with monospecific antibodies indicated that 3 of the 4 lines expressed simple epithelial keratins 8 and 18, whereas 1 of the 4 also expressed keratin 19. A panel of lectins revealed characteristic localization patterns distinct from those observed on other epithelial cell lines. Cells from 3 lines (TR131, TR138, and TR146) inoculated into nude mice (nu/nu) produced cystic nodules or unequivocal tumors having a histology indicating a squamous cell origin for the injected cells. Electron microscopy demonstrated that the cell lines covered a spectrum of differentiation capability ranging from the undifferentiated monolayer cultures of TR126 to the rather well differentiated, stratified cultures of TR131.

Adult↗
PubMed1985

The microenvironment of coeliac disease: T cell phenotypes and expression of the T2 'T blast' antigen by small bowel lymphocytes.

Peroral jejunal biopsies were studied by double-label immunofluorescence on cryostat tissue sections from control patients of normal histology and patients with coeliac disease. A panel of monoclonal antibodies was used to identify T cells and T cell subsets, together with antibody markers of cell stimulation (the T2 40 kdalton T blast specificity), proliferation (T9 and Ki67) and activation (HLA-DR and Ig receptors). In normal mucosa, expression of T2 was predominantly found in the T8+ intraepithelial cytotoxic/suppressor population. In coeliac disease there was little alteration of T4:T8 ratios per se, but a much higher percentage of T8-T4+ helper/inducer cells expressed the T2 antigen (approx. 30% compared to approx. 10%), manifest as an accretion of T2+T8- cells in the epithelium and subjacent stroma. Additionally, T cell lymphoid aggregates were observed in the lamina propria, consisting of greater than 90% T4+ cells, of which the majority (60-80%) were also T2+. The increase of stimulated helper cells correlated well with the pathology of coeliac disease, being most marked in untreated cases of maximal tissue damage, and least in well treated cases with restoration of normal morphology. By contrast, an increased tendency for the T8+ cells to co-express T1 in untreated coeliac disease was not observed in treated disease, even in cases where failure to adhere to a strict gluten free diet had resulted in little histological improvement. There was little evidence of T cell activation or proliferation. The observation that T helper cells undergo migration and stimulation suggests an imbalance in immunoregulation which causes a breakdown of normal immune tolerance to dietary gluten, and therefore may be a crucial mechanism of tissue damage in coeliac disease.

Adolescent↗
PubMed1985

Microenvironments in the normal thymus and the thymus in myasthenia gravis.

The disposition of epithelial cells and extracellular matrix, in the thymus of 8 cases of myasthenia gravis (MG) and in controls (over a wide age range) was studied. In the controls, the subcapsular epithelium was strongly Leu-7-positive in the fetus, negative in childhood, and positive again in adults. Another antibody, RFD4, also labeled the subcapsular epithelium in childhood and adults, but not fetal samples. The samples from MG cases showed the same staining pattern as adult control samples. The medullary epithelium was also RFD4+, and at all ages. The most striking changes in the advanced cases of MG were the unusual arrangement and hypertrophic appearance of medullary epithelial cell areas, separated by laminin-positive basement membranes from the alternating multiple bands of peripheral lymph-node-like areas. The latter had regions resembling the paracortex of lymph nodes as well as germinal centers (GCs). The T-cell zones contained heavy deposits of fibronectin. These T-cell zones were unique to the thymus in MG and were absent in the two normal thymic samples with isolated GCs. In MG the laminin-containing basement membrane, which separated the medullary epithelial and peripheral lymph-node-like areas, was fenestrated at circumscribed points closest to the GCs, thus apparently permitting communication among the medullary epithelium, the T-cell zones, the GCs and the associated antigen-presenting cells. Large numbers of interdigitating cells and some lymphocytes of cortical thymocyte phenotype were also found at these special sites, where opportunities for autosensitization may persist in MG.

Adult↗
PubMed1985

Human B cell development. I. Phenotypic differences of B lymphocytes in the bone marrow and peripheral lymphoid tissue.

The phenotype of B lineage cells (TdT+, pre-B, IgM+, IgD-, and IgM+,IgD+) in infant and adult human bone marrow was compared with that of B cells seen in peripheral tissues such as tonsil and blood. The range of B cell-associated antibodies used included four reagents with greater than 90% reactivity on peripheral B cells: RFB4 and To15 (both p135, corresponding to CD22), RFB6 (p140 corresponding to CD21), and Y29/55, a unique B cell-specific antibody. In addition, AL-1, an antibody with virtually no reactivity against peripheral B cells was also used. The BM cell subpopulations were heterogeneous in respect of antibody reactivity. The TdT+, pre-B and IgM+, IgD- cells were AL-1+ but did not express membrane antigens recognized by the antibodies To15, RFB4 (CD22), and RFB6 (CD21). TdT+, pre-B cells, and 50% of IgM+, IgD- BM B cells were also unreactive with antibody Y29/55, the other 50% being Y29/55+. In contrast, the IgM+,IgD+ BM B cells, like peripheral B cells, were positive with antibodies To15, RFB4, RFB6, and Y29/55, but reacted only in small numbers with AL-1. The orderly differentiation-linked display of these antigens was also suggested by the findings that normal TdT+, pre-B, and IgM+,IgD- cells expressed the To15 and RFB4 (CD22) antigens in their cytoplasm (in the Golgi region). This observation was confirmed in malignant common acute lymphoblastic and pre-B blast cells, as well as in the corresponding permanent cell lines KM3 and NALM-6. In these lines the membrane expression of To15 and RFB4 could be induced by phorbol ester during a 48 to 72 hr culture period.

Adolescent↗
PubMed1985

An ultrastructural analysis of the physical organization of collagenous (type I) matrices: one determinant of urothelium maintenance in vitro.

Collagenous matrices, used as cell culture substrata, can be prepared from different collagen types in a variety of forms using a range of polymerization procedures. Type I collagen has been most frequently used either as dried collagen films or hydrated collagen gels. Sheets of isolated bladder urothelium, when plated onto such matrices prepared from type I collagen by different polymerization methods (eg. air-drying; NaOH; NaCl; NH3; or NH3 followed by glutaraldehyde crosslinking) demonstrate the capability of urothelial cells to attach to a variety of differently prepared matrices irrespective of polymerization procedure. In contrast, both cell proliferation and maintenance of the urothelium are markedly influenced by the polymerized form of the collagen matrix. Comparative ultrastructural (scanning and transmission electron microscopy) analysis of these matrices demonstrates dissimilarities in their physical organization. The level of filamentous, fibrillar or fibrous reaggregation of solubilized collagen molecules varies in relation to the polymerization procedure used viz, a) air dried matrices form a dense meshwork of many forms of collagen fibrils and associated filaments with an irregular surface array of coarser collagen fibres; b) matrices prepared by NaOH, NaCl and NH3 polymerization present no major differences and form a felt of interlocking collagen fibres with discrete filamentous networks associated with these fibres; and c) matrices polymerized by NH3 and crosslinked with glutaraldehyde form a dense meshwork of filaments with a more occasional distribution of fibrils associated with filaments or dense "amorphous" aggregates. The level of supramolecular reassemblage of solubilized collagen may be, therefore, a significant factor in determining urothelial cell growth and differentiation on collagen matrices.

Animals↗
PubMed1984

Depletion of T lymphocytes in donor marrow prevents significant graft-versus-host disease in matched allogeneic leukaemic marrow transplant recipients.

For more than 15 years preclinical studies have suggested that acute graft-versus-host disease (aGvHD) might be prevented by the removal of immunocompetent T lymphocytes from the donor marrow inoculum. To test this observation in man 14 patients were given marrows virtually (greater than 99%) depleted of identifiable donor marrow T lymphocytes by the use of a "cocktail" of specific anti-T-cell monoclonal antibodies (MBG6 and RFT8) and rabbit complement. Patients were not given immunosuppressive prophylaxis after bone-marrow transplantation. Moderate to severe (grades II-IV) GvHD was totally prevented. 2 of 13 evaluable patients showed mild (grade I) skin GvHD only. Although peripheral blood recovery was slower than that obtained with other forms of GvHD prophylaxis, no fatal infections occurred. All patients survived the early post-transplant period.

Acute Disease↗