Biomedical subjects

H Gamliel

Publications and source records attributed to H Gamliel.

At least 37 records · Page 2Linked to original sources

PubMed1983

Utilization of monoclonal antibodies and immuno-scanning electron microscopy for the positive identification of human leukemic cells.

Monoclonal antibodies generated against normal and leukemic human leukocytes were tested for their differential reactivity with leukemia and lymphoma cell lines as well as with circulating lymphoid and myeloid leukemic cells by means of immuno-scanning electron microscopy (immuno-SEM). Anti-T (OKT3), anti-mu-chain, anti-CALLA (J5), anti-BA-1, anti-BA-2, and anti-nonlymphoid (Mol) monoclonal antibodies were covalently conjugated to polystyrene latex microspheres (immunolatex), using a two-step glutaraldehyde reaction, and subsequently incubated with the various cell types. Cultured B-type Burkitt lymphoma cells (Daudi) and chronic lymphocytic leukemia (CLL) cells displayed extensive labeling with monoclonal anti-mu, anti-B1, and anti-BA-1 immunolatex conjugates, while cultured malignant T cells (HD-Mar) showed positive labeling with OKT3 immunolatex alone. Cultured myelomonocytic cells (GDM-1) and cells obtained from patients with acute myeloblastic (AML) and monoblastic leukemia (AMoL) labeled only with anti-Mol immunolatex, while cultured promyelocytic cells (HL-60) displayed far less labeling with this conjugate. Common-type acute lymphoblastic leukemia (C/ALL) cells were labeled predominantly with the J5 (anti-CALLA) and anti-BA-2 immunolatex conjugates. Evidence is presented indicating that immuno-SEM employing monoclonal antibodies is a reproducible technique which may be used in the study of leukocyte maturation and may provide additional information in the classification of poorly differentiated leukemias.

Antibodies, Monoclonal↗

PubMed1983

Air-drying of human leucocytes for scanning electron microscopy using the GTGO procedure.

The utilization of tannic acid and guanidine hydrochloride as mordants for better osmium binding has been shown to serve as an excellent alternative to metal coating of organ tissue specimens for scanning electron microscopy (SEM). The present report describes the GTGO procedure, a modification of the TAO technique introduced by Murakami et al. (1977, 1978), which we have found successful for the preparation of air dried peripheral blood leucocytes for SEM studies. Air dried, GTGO-treated leucocytes show excellent preservation of surface features with minimal cell shrinkage. When critical point dried, GTGO-treated cells are examined, they also show less shrinkage than cells prepared with standard glutaraldehyde fixation and critical point drying. The potential application of this air drying procedure (GTGO-AD) to other soft biological specimens is currently under investigation. This technique is recommended as a new and effective air drying procedure for the successful preparation of cells for SEM.

Freeze Drying↗

PubMed1983

Surface features of leukaemic megakaryocytic precursors. A study of 5 cases of megakaryoblastic leukaemia with scanning electron microscopy.

Megakaryoblasts and maturing megakaryocytic precursor cells from 5 patients with megakaryoblastic leukaemia were studied by scanning electron microscopy (SEM). The diagnosis in all cases had been established by ultrastructural cytochemistry on the basis of a positive platelet peroxidase reaction with negative staining for myeloperoxidase. 1 case presented as acute myelofibrosis and 4 as acute megakaryoblastic transformation of chronic granulocytic leukaemia. Under the SEM, megakaryoblasts and maturing megakaryocytic precursors showed typical surface features including the presence of rounded and irregular blebs, broad folds and pseudopodia. The nature of these surface blebs is still unclear but they probably represent surface membrane alterations relating to imminent platelet shedding at least in the more mature precursors. These surface microprojections are distinctly different from those encountered on leukaemic lymphoblasts, myeloblasts and monoblasts. It is suggested that SEM may be used in conjunction with the PPO reaction as in aid in the diagnosis of megakaryocytic leukaemias.

Blood Platelets↗

PubMed1983

Scanning immuno-electron microscopy of a monoclonal, Epstein-Barr Virus(EBV) transformed human cell line producing rheumatoid factor in-vitro.

Selected lymphocytes from a patient with rheumatoid arthritis were infected with Epstein-Barr virus (EBV) and the emerging cell line (RF-AN) has now been successfully maintained in culture for more than two years. Multiparameter studies, including surface markers, ultra-structure and scanning immuno-electron microscopy, were utilized to characterize and evaluate this unique human cell line. Like other B-lymphoblastoid cell lines, RF-AN cells display multiple microvilli, contain surface and intracytoplasmic immunoglobulin (IgM lambda) and continuously produce monoclonal IgM lambda in vitro. The latter serves as an autoimmune antibody directed against IgG and is termed rheumatoid factor (RF). When RF-AN cells were incubated with human or rabbit IgG conjugated to polystyrene (latex) microspheres, almost all cells were labeled with the marker. However, cells did not label with goat IgG conjugated to latex. RF-AN cells were also labeled with latex microspheres coupled to goat antisera directed against human IgM or lambda chains but not with goat anti-human IgG or anti-human kappa chains. It is suggested that this mode of immunomicroscopy is reliable and may provide more useful data than other standard immunological techniques.

Antibodies, Monoclonal↗

PubMed1983

Cell surface alterations induced by the acycloguanosine (ACG) resistant and sensitive strains of herpes simplex virus type 2 (HSV-2) in the presence of ACG and tunicamycin.

HSV-2-ACG, a mutant of herpes simplex virus type 2, which is resistant to acycloguanosine and also capable of inducing fusion of BSC1 cells, was isolated. The viral-induced surface changes in these cells were studied using scanning electron microscopy. The formation of gigantic fused cells with increased number of microvilli and virus-like particles, extensively covering the infected cell surface, were investigated. Tunicamycin (50 micrograms/ml), an inhibitor of glycosylation of polypeptides, blocks the production of infectious virus, as well as the cell-fusion and efficiently prevents the viral induced alterations occurring on the surface of the cells.

Acyclovir↗

PubMed1983

The use of scanning immuno-electron microscopy to detect surface membrane immunoglobulins and antigens on normal and leukemic human leukocytes: current status.

In this report we review the use of scanning immuno-electron microscopy (SIEM) as a means of elucidating the location of cell surface membrane immunoglobulins (SMIg) and antigens (SMAg) on normal and leukemic human leukocytes. SMIg as well as SMAg were detected on various subpopulations of cells under the SEM, readily enabling identification of different types of circulating and cultured leukocytes. It is apparent from current data that the application of highly-specific monoclonal antibodies, multiple labeling procedures and improved methods of quantitation with SEM may well provide a more sensitive and elegant methodology for the identification of individual membrane components in a wide variety of mammalian cell systems.

Antigens, Surface↗

PubMed1982

Hybridization between a human epithelial line, infectable by Epstein-Barr virus, and Burkitt lymphoma lines: membrane properties, superinfectability, inducibility and tumorigenicity.

The human epithelial line U, which is partially infectable with EBV, was hybridized with the EBV-genome carrying Burkitt lymphoma lines P3HR-1 and Daudi. Authenticity of the hybrids U-Put and U-Dut was established by isoenzyme studies. Although the two hybrids carried the EBV genome derived from the lymphoma parent, being 100% positive for Epstein-Barr-virus-associated nuclear antigen (EBNA), they resembled the U parent in many respects: they were deficient for membrane immunoglobulins and Fc receptors, and had a lower concentration of EBV-C3 receptors than either parent. Unlike the P3HR-1 parent, U-Put hybrid was nonpermissive for both the EBV cycle antigens, early antigen (EA) and viral capsid antigen (VCA). The inducing agent 12-O-tetra-decanoyl-phorbol-13-acetate (TPA) caused distinct viral early antigen synthesis (EA) in U-Put, lower, however, than that of the parental P3HR-1. U-Dut was completely nonpermissive and noninducible for early and viral capsid antigens. Thus, even an epithelial parent infectable by EBV restricted, although not completely, expression of EBV antigens, with the exception of EBNA. It has been suggested that EBNA is an autonomous function of the viral genome, independent of host cell control; the latter regulates expression of antigens related to viral cycle. The hybrids U-Put and U-Dut resembled the U parent also in regard to growth in soft agar and tumorigenicity in nude mice, although in this respect the lymphoma parental properties were not completely eclipsed.

Animals↗

PubMed1982

Exposure to phorbol diester (TPA) in vitro as an aid in the classification of blasts in human myelogenous and lymphoid leukemias: in vitro differentiation, growth patterns, and ultrastructural observations.

Leukemic cells from the peripheral blood of 52 patients with acute and chronic leukemias were incubated with 12-0-tetradecanoyl phorbol ester (TPA). Thirty-one cases of lymphocytic leukemia (18 cases of acute lymphoblastic and 13 cases of chronic lymphocytic leukemia), 13 cases of acute nonlymphoblastic (myelo or myelomonoblastic) leukemia, and eight cases of blastic crisis of CGL (seven cases of predominantly myeloblastic crisis, and one case of lymphoblastic crisis) were studied. In all cases of lymphoid leukemia, cells formed clumps or aggregates after exposure to TPA, while in all cases of myeloid leukemia cells became adherent to the substrate. Seven of the eight cases of blastic crisis of CGL were predominantly myeloid in type and cells adhered to the substrate, while in a single case of lymphoid crisis in CGL cells formed clumps after TPA exposure. Functional, cytochemical, and ultrastructural studies showed altered cell differentiation and continuing in vitro maturation of leukemic cells after exposure to TPA. In the light of the above results, it is concluded that this simple test employing TPA exposure in vitro serves as a reliable means of distinguishing blasts from different origins in human leukemias.

Histocytochemistry↗

PubMed1982

Simultaneous presentation of plasma cell and monocytic leukemia with a subacute clinical course.

A rare case of simultaneous presentation of monocytic and plasma cell leukemia is reported. Cytochemistry, transmission and scanning electron microscopy confirmed the presence of a dual population consisting of monoblasts and plasma cells. Monoblasts contained nonspecific esterases, secreted lysozyme and showed dense bodies and surface ruffles under the scanning electron microscope, while the plasma cells secreted IgG kappa paraprotein, contained rough endoplasmic reticulum, and showed surface blebs with microvilli. Another unusual feature of this case was the relatively chronic course of the disease, lasting 15 months after initial diagnosis.

Acute Disease↗

PubMed1982

Metabolic and ultrastructural aspects of the in vitro lysis of chronic lymphocytic leukemia cells by glucocorticoids.

Human chronic lymphocytic leukemia (CLL) cells like prothymocytes and immunoactivated T-lymphocytes are readily lysed in vitro by pharmacological concentrations of glucocorticoids such as cortisol, whereas peripheral blood lymphocytes and thymocytes are unaffected by the hormone. In this study, metabolic and ultrastructural aspects of the cortisol-induced killing process of CLL cells are recorded. In vitro lysis was found to be temperature dependent and was detected only after 6 to 8 hr incubation with cortisol by means of the trypan blue exclusion test. However, 30 min of incubation with cortisol at either 37 degrees or 4 degrees followed by the removal of the hormone was still sufficient to induce the lytic process. Ultrastructural studies demonstrated sequential changes in the cytoplasm, including swelling of mitochondria and cytoplasmic decompartmentalization, followed by loss of surface microvilli with the appearance of "holes" in the cell membrane, and subsequent condensation of nuclear chromatin. The large holes in the membrane appearing after 6 hr of incubation with the hormone may be the cause for the penetration of the viable stain into the dead cells, as seen by light microscopy. Addition of metabolic inhibitors including actinomycin D, puromycin, and cycloheximide following administration of cortisol resulted in inhibition of the cell lysis. An excess of an antagonist such as cortexolone was found to inhibit the cortisol-induced cytolysis of the CLL cells. It is suggested that the glucocorticoid-induced lysis of human CLL cells is similar to the phenomenon observed in rat or murine lymphocytes and is mediated by interaction of the steroid molecule with the cytoplasmic receptor. The resulting complex appears to activate specific gene(s) the products of which eventually cause cytolysis.

Dactinomycin↗

PubMed1981

Plasma cell leukemia and myeloma: a scanning electron-microscopic study of cell surface features in six cases.

Circulating plasma cells from six patients who had plasma cell leukemia were examined by transmission and scanning electron microscopy. In all cases, leukemic plasma cells constituted more than 60% of the total cell population in the peripheral blood. Transmission electron microscopy confirmed that the leukemic cells were plasmacytic and that many of them contained parallel arrays of rough endoplasmic reticulum and a prominent Golgi apparatus. Scanning electron microscopy confirmed previous observations of cultured myeloma cells and showed that plasma cells display varying numbers of surface blebs in addition to short stublike microvilli. The microvilli were frequently clustered together in one area of the surface. Bleb formation appears to be characteristic of plasma cells, but its nature is still obscure. Current knowledge of this phenomenon is briefly reviewed.

Adult↗

PubMed1981

Scanning immuno-electron microscopy of human leukaemia and lymphoma cells: a comparative study of techniques using immunolatex spheres as marker.

In this study scanning immuno-electron microscopic (SIEM) techniques were used to identify human leukaemia-lymphoma cells. Monodispersed polystyrene (latex) beads were conjugated to specific antisera using glutaraldehyde, in an attempt to detect surface antigenic components on a variety of cells of known origin. Antisera, mostly immunoglobulin fractions, against human thymus (T) derived cells, common type acute lymphoblastic leukaemia cells (C/ALL) and surface immunoglobulin (sIg) bearing cells were used to coat latex spheres, while rabbit anti-mouse Thy-1 antiserum or whole human-IgG (gamma-globulin) bound to latex were used as controls in some experiments. The use of SIEM techniques in the direct mode as a simple and sensitive method for labelling surface antigens is described. The disadvantages of the SIEM methodology are also summarized while the requirements for optimal cell preparation using this technique are stressed. The experiments were designed to ascertain whether prolonged fixation of cells could be used prior to incubation of the cells with the marker. In this respect, repeated neutralization of the glutaraldehyde with glycine is essential. SIEM labelling of cells is random and unreliable without adequate quenching with glycine. The heteroantisera used in this study proved to be adequate and insignificant non-specific attachment and cross reactivity were seen. SIEM adds a further dimension to ultrastructural aspects of immunology and is a potentially useful tool in the study and identification of leukaemia and lymphoma cells.

Antigens, Surface↗

PubMed1981

Positive identification of human leukaemic cells with scanning immuno-electron microscopy, using antibody coated polystyrene (latex) beads as markers.

Scanning immuno-electron microscopic labelling techniques were successfully used to identify a variety of leukaemia-lymphoma cells. Anti-T, -B and -common type acute lymphoblastic leukaemia (ALL) antisera were covalently conjugated to polystyrene latex spheres by means of glutaraldehyde. Peripheral blood mononuclear cells obtained from healthy individuals and from patients with T-type ALL, B-type chronic lymphocytic leukaemia (CLL), acute and chronic non-lymphoblastic leukaemias (ANLL) and cultured leukaemia-lymphoma cells were incubated with the above antisera-latex conjugates. Positively labelled cells were easily identified by the presence of latex spheres on their surfaces. Cross reactivity of the antisera used and non-specific attachment of the markers were infrequently encountered. The results of this study confirm earlier scanning electron microscopic reports indicating that different leukaemic cells have characteristic surface features and show that scanning immuno-electron microscopy (SIEM) can readily be applied to the study of human leukaemic cells.

Humans↗

PubMed1981

Use of multiparameter studies and scanning electron microscopy in the interpretation and attempted correlation of surface morphology with cell type in 135 cases of human leukemias.

Multiparameter studies and scanning electron microscopy (SEM) were performed on cells obtained from 135 cases of leukemia in an attempt to clarify whether there was a reliable correlation between surface morphology and cell type as defined by cytochemistry, membrane markers, and transmission electron microscopy. These studies also attempted to determine whether SEM could be used to distinguish lymphoid and nonlymphoid leukemias, to recognize different types of lymphoid leukemia, and to define the cell type involved in cases of unclassified leukemia. The results of this study suggest that there is a good correlation between surface morphology as seen by SEM and cell type identified by multiparameter techniques. In most cases, nonlymphoid leukemic cells could be distinguished from lymphoid leukemic cells on the basis of their surface morphology. SEM did not appear to contribute to the diagnosis of unclassified leukemia, but more cases of this nature must be studied. Despite the fact that acute lymphoblastic leukemia cells frequently showed fewer microvilli than did other lymphoid leukemias, overlap of surface features in about one-third of the cases did not enable SEM to be used as a reliable means of distinction. The above conclusions appear to be supported by preliminary scanning immunoelectron microscopic observations on leukemic cells. It is concluded that SEM is a useful aid to other modes of microscopy in leukemia but should not be used on its own to establish diagnosis.

Cell Membrane↗

PubMed1981

Ultrastructural, cell membrane, and cytogenetic characteristics of B-cell leukemia, a murine model of chronic lymphocytic leukemia.

A murine model of a spontaneous, transplantable BALB/c B-cell leukemia (BCL1) is described. Extreme leukemia and splenomegaly develop in H-2d-compatible recipients of tumor cells. Tumor cells are medium to large lymphocytes that can be transformed into plasmacytoid cells following in vitro stimulation with lipopolysaccharide. Karyotypic analysis of transformed tumor cells reveals 36 chromosomes with several monosomies and 7 markers chromosomes. The ultrastructure of the tumor cells was studied using transmission and scanning electron microscopy. Although the appearance of tumor cells seems normal by morphological criteria, an impaired capping ability was documented using the fluorescein-conjugated concanavalin A-binding test. Impaired capping ability was documented before leukemia was overt as early as 1 to 3 days following inoculation of tumor cells. The B-cell leukemia (BCL1) provides a useful murine model for the study of various aspects of human bone marrow-derived malignant disorders.

Animals↗

PubMed1980

Human prothymocytes. Membrane properties, differentiation patterns, glucocorticoid sensitivity, and ultrastructural features.

Thymic precursor cells (prothymocytes) comprise a large proportion of the fetal thymic cell population, but are less frequently encountered in the postnatal thymus, where they compose < 1% of the entire population. In the present study we attempted to characterize a number of properties of the prothymocytes obtained from human fetal thymic tissues after depletion of the E-rosette thymocyes on a Ficoll-Hypaque gradient. The prothymocytes are larger than the thymocytes and show a different nuclear chromatin pattern. This subset of cells lacks the E-rosetting and natural-attachment capacities and, unlike thymocytes, does not bind the lectin peanut agglutinin. Human prothymocytes are highly sensitive to the in vitro cytolytic effect of hydrocortisone, whereas the thymocytes are resistant. Long-term in vitro culture of prothymocytes resulted in the expression of thymocyte characteristics together with a burst of mitotic activity. Results of this study indicate that the rate of the prothymocyte proliferation is regulated by the small thymocytes present in the same suspension.

Antigens, Surface↗

PubMed1980

Establishment in continuous culture of a T-lymphoid cell line (HD-Mar) from a patient with Hodgkin's lymphoma.

A new cell line, HD-Mar, was established from a pleural effusion of a patient with Hodgkin's lymphoma. Formation of E rosettes, sensitivity to anti-T serum, elevated terminal deoxynucleotidyl transferase activity, presence of T-cell and the common ALL membrane antigens, morphology, and cytochemical staining indicate that the HD-Mar line is of thymic derivation. Absence of any immunoglobulin determinants, the lack of EBNA or any other EBV-associated antigen or function are also characteristics associated with established T-cell-derived lymphoma cell lines. Karyotype analysis indicated a tetraploid origin of the cell line.

Adult↗

PubMed1980

Scanning electron microscopy of mycoplasmas adhering to erythrocytes.

The interaction of Mycoplasma pneumoniae and Mycoplasma gallisepticum with human erythrocytes (RBC) was studied by scanning electron microscopy. The tight nature of the attachment of the microorganisms to the RBC was indicated by the indentation of the RBC surface at the site of attachment of M. gallisepticum cells and by traction and resulting distortion in the shape of the RBC at the point of its attachment to M. pneumoniae filaments growing on glass or plastic. In many cases attachment took place via the tip of the filaments, the membrane of the parasite appearing to be fused with that of the RBC. The morphology of the mycoplasmas growing on cover slips conformed in general with previous descriptions obtained by scanning electron microscopy. Growth of M. pneumoniae on glass or plastic consisted of branching filaments spread on the inert surface and microcolonies made up of intertwining filaments projecting into the medium. The filaments had a bulbous swelling adjacent to a tapered tip end. A few filaments were shown to have a ropelike helical twist. M. gallisepticum grown on the cover slips of Leighton tubes had a peculiar fusiform or teardrop shape with blebs at one or both poles of the cells. Elongated filamentous forms and chains of coccobacillary bodies were observed as well.

Cell Adhesion↗