Biomedical subjects

K L Sung

Publications and source records attributed to K L Sung.

At least 37 records · Page 2Linked to original sources

PubMed1994

Interleukin-1 beta induces differential adhesiveness on human endothelial cell surfaces.

The adhesive strength between HL-60 and endothelial cells activated with IL-1 beta was investigated according to the region (area) on the endothelial cell surface where the HL-60 cell was attached (nuclear, junctional, or cytoplasmic). Using a micropipette single-cell manipulation system, we demonstrated that the increase of adhesive force was due to the activation of the endothelial cell with IL-1 beta and a function of the region of the endothelial cell. The increase in adhesion strength due to the increased expression of E-Selectin on the endothelial cell was studied by the addition of monoclonal antibodies against E-Selectin, ICAM-1, VCAM, and PECAM-1. Our findings suggest that the greatest adhesive strength was seen in the junctional region where the antibodies could not block the adhesion. However, the contribution of E-Selectin was significant in the nuclear region.

Antibodies, Monoclonal↗

PubMed1994

Adhesion strength of human ligament fibroblasts.

The adhesive interactions of cells with other cells and the extracellular matrix (ECM) play a fundamental role in the organization of cells in differentiated organs, cell motility, and the healing process. The adhesion characteristics of ligament fibroblasts depend on the expression of cell surface molecules and their interaction with the ECM. Although many receptors mediating the effects of ECM components on ligament cell function remain poorly defined, it is known that fibronectin (FN) allows ligament cells to adhere through the VLA-5 receptor (alpha 5 beta 1). A direct measurement of the adhesion between anterior cruciate ligament (ACL) or medial collateral ligament (MCL) fibroblasts and fibronectin matrix proteins was achieved by using a micromanipulation technique to determine the force required to detach an ACL or MCL cell from fibronectin-coated glass. We have found that the adhesion strength is not random, but has well-defined functional relationships with the FN concentration and the seeding time (time allowed for the cell to establish attachment). The adhesion strength (i.e., force required to detach) of ACL cells shows a stronger dependence on FN concentration (1, 2, and 5 micrograms/ml) for short seeding times (15-30 min) than for long seeding times (38-75 min). For MCL cells, the effect of the seeding time on adhesion strength was apparent for all concentrations. For all the seeding times studied and FN concentrations used, MCL cells had higher adhesion strength than ACL cells. The adhesion strengths of ACL and MCL fibroblasts to FN are correlated to cell adhesion area.(ABSTRACT TRUNCATED AT 250 WORDS)

Anterior Cruciate Ligament↗

PubMed1993

Determination of adhesion force between single cell pairs generated by activated GpIIb-IIIa receptors.

A biophysical approach was used to directly determine the avidity of the junction between two Chinese hamster ovary (CHO) cells bearing recombinant GpIIb-IIIa in the presence and absence of fibrinogen. Micromanipulation was used to induce conjugation of the cell pairs with or without activating the GpIIb-IIIa molecules with monoclonal antibody (MoAb) 62. Activation of GpIIb-IIIa caused an increase in the force required to separate the conjugates. The molecular bonding force between cells bearing activated GpIIb-IIIa and fibrinogen molecules was found to be 2.1 x 10(-7) dyne, which is 3.7 times higher than that between nonactivated GpIIb-IIIa and fibrinogen (5.7 x 10(-8) dyne). The results provide a quantitative assessment of the molecular bonding force between fibrinogen and the GpIIb-IIIa expressed on cell surface. The findings indicate that the activation of GpIIb-IIIa leads to an increase in the adhesive force in CHO cell aggregation by increasing the strength of the GpIIb-IIIa-fibrinogen bonds rather than the number of these bonds.

Animals↗

PubMed1993

Human monocyte colony-stimulating factor stimulates the gene expression of monocyte chemotactic protein-1 and increases the adhesion of monocytes to endothelial monolayers.

The stimulation of the human umbilical vein endothelial cell (HUVEC) with recombinant human monocyte-derived colony-stimulating factor (MCSF) increased the gene expression of monocyte chemotactic protein (MCP-1). Northern blot analysis indicated that 50 U/ml of MCSF is the optimal concentration for this effect. The elevation of MCP-1 mRNA started as early as 1 h after stimulation and was maintained for at least 8 h. An increased MCP-1 level in MCSF-treated HUVEC was also demonstrated at the protein level by immunocytochemical staining using a polyclonal MCP-1-specific antibody. HUVEC activated by 50 U/ml of MCSF for 5 h showed a stronger immunofluorescence staining than control cells. Micropipette separation of THP-1 monocytes from HUVEC showed that the activation of both THP-1 and endothelium by MCSF led to an increase in the separation force by more than three times (36.2 +/- 6.7 x 10(-4) vs. 9.6 +/- 3.6 x 10(-4) dyn). An increased adhesiveness was also observed after MCSF activation of peripheral blood monocytes and HUVEC (16.7 +/- 2.7 x 10(-4) vs. 5.2 +/- 0.9 x 10(-4) dyn). The increased adhesive force in both systems was blocked by the use of anti-MCP-1 (5.5 +/- 0.8 x 10(-4) and 6.8 +/- 1.1 x 10(-4) dyn). Similar results were obtained in experiments in which only HUVEC, but not monocytes, were activated by MCSF. This increased adhesion of untreated monocytes to MCSF-activated HUVEC was also blocked by the addition of anti-MCP-1. In contrast, experiments in which only THP-1 or peripheral blood monocytes, but not HUVEC, were treated with MCSF did not show a significant increase of adhesion between these cells. These results indicate that MCSF augments monocyte-endothelium interaction primarily by its action on the endothelial cell and that this function is probably mediated through an increased expression of MCP-1. The MCSF/MCP-1-dependent adhesive mechanism might be operative in the arterial wall in vivo to lead to the trapping of the infiltrated monocyte-macrophage in the subendothelial space during atherogenesis.

Antibodies↗

PubMed1992

Micromanipulation of adhesion of a Jurkat cell to a planar bilayer membrane containing lymphocyte function-associated antigen 3 molecules.

Cell adhesion plays a fundamental role in the organization of cells in differentiated organs, cell motility, and immune response. A novel micromanipulation method is employed to quantify the direct contribution of surface adhesion receptors to the physical strength of cell adhesion. In this technique, a cell is brought into contact with a glass-supported planar membrane reconstituted with a known concentration of a given type of adhesion molecules. After a period of incubation (5-10 min), the cell is detached from the planar bilayer by pulling away the pipette holding the cell in the direction perpendicular to the glass-supported planar bilayer. In particular, we investigated the adhesion between a Jurkat cell expressing CD2 and a glass-supported planar bilayer containing either the glycosyl-phosphatidylinositol (GPI) or the transmembrane (TM) isoform of the counter-receptor lymphocyte function-associated antigen 3 (LFA-3) at a concentration of 1,000 molecules/microns 2. In response to the pipette force the Jurkat cells that adhered to the planar bilayer containing the GPI isoform of LFA-3 underwent extensive elongation. When the contact radius was reduced by approximately 50%, the cell then detached quickly from its substrate. The aspiration pressure required to detach a Jurkat cell from its substrate was comparable to that required to detach a cytotoxic T cell from its target cell. Jurkat cells that had been separated from the substrate again adhered strongly to the planar bilayer when brought to proximity by micromanipulation. In experiments using the planar bilayer containing the TM isoform of LFA-3, Jurkat cells detached with little resistance to micromanipulation and without changing their round shape.

Antigens, CD↗

PubMed1992

Granulocyte adhesion, deformability, and superoxide formation in acute stroke.

BACKGROUND AND PURPOSE: Impaired rheological properties of as well as cytotoxic substances produced by granulocytes may contribute to tissue damage in acute ischemic stroke. To assess changes in the properties of circulating granulocytes, we measured their adhesion, deformability, and superoxide generation in the first 3 days after ischemic stroke. METHODS: Granulocytes from 18 male patients and 20 age- and risk-matched controls were investigated. Adhesion to murine laminin-, human fibronectin-, and bovine serum albumin-coated surfaces was measured with and without the stimulus phorbol myristate acetate and the antiadhesion antibody IB4. Superoxide anion formation was assessed by the reduction of ferricytochrome C. In a subgroup of 10 patients and 11 controls, granulocyte deformability was determined using the micropipette aspiration technique. RESULTS: The patients had significantly greater granulocyte adhesion to laminin (p less than 0.005) and fibronectin (p less than 0.05) but not bovine serum albumin. Cell stimulation enhanced the differences between the groups, whereas the antiadhesion antibody inhibited adhesion in both patients and controls. There were no significant differences in granulocyte deformability. Superoxide production by granulocytes was significantly lower in the patients without the stimulus (p less than 0.05) and with 10 nM phorbol myristate acetate (p less than 0.005). CONCLUSIONS: These findings suggest that circulating granulocytes in ischemic stroke exhibit increased adhesive properties, a feature that represents one of the risk factors for granulocyte entrapment, impairment of microvascular flow, and tissue injury.

Acute Disease↗

PubMed1992

Force contribution of the LFA-1/ICAM-1 complex to T cell adhesion.

Little is known in quantitative terms about forces between cells generated during adhesion and recognition, or about the contribution of any one set of molecular associations to the development of these forces. To determine the forces involved in adhesion dependent on lymphocyte function-associated antigen-1 (LFA-1) and intercellular adhesion molecule 1 (ICAM-1), we have measured the junctional avidity between single cell pairs consisting of a cloned T cell that expresses LFA-1 and a fibroblast cell that expresses MHC class II molecules and ICAM-1 after transfection. Micromanipulation was used to induce conjugation of cell pairs and to determine the force required to separate the conjugate. T cell adhesion to three related fibroblast cell lines was compared: the parent line that does not express ICAM-1 or other LFA-1 counter-receptors, and two transfectants that have high and moderate levels of surface ICAM-1 expression. The force needed to separate the conjugates varied with the fibroblast ICAM-1 expression levels. The T cell adhesion to ICAM-1-expressing fibroblasts was strong, and the critical separation stresses measured for the three cell lines were 1.4 x 10(3) dyn/cm2 (1 dyn=10(-5) N) for the ICAM-1-negative fibroblast, 4.98 x 10(3) dyn/cm2 for the fibroblast with a moderate level of ICAM-1 expression, and 6.25 x 10(3) dyn/cm2 for the fibroblast line with the highest ICAM-1 expression. The dependence of adhesion strength on the LFA-1/ICAM-1 complex was confirmed by the use of blocking antibodies, which showed the contribution from the interaction of CD4/MHC class II to be negligible.

Animals↗

PubMed1992

Influence of temperature on rheology of human erythrocytes.

The effect of variation in temperature on the rheological properties of human red blood cells was studied by determining the cell deformation in response to aspiration pressure applied via a micropipette. The time history of the deformation was analyzed by the use of a Voigt viscoelastic model consisting of an elastic element (E) in parallel with a viscous element (eta). Viscosity values were obtained during the initial rapid deformation phase (phase I), the later slow deformation phase (phase II), and the recovery phase. With a rise in temperature from 0 to 45 degrees C, both E and eta values decreased. A thermotropic transition temperature was found at approximately 21 degrees C for phase I viscosity and at approximately 9 degrees C for viscosity in the recovery phase, but not for phase II viscosity. These responses to temperature variations provide insights into the viscoelastic properties of the erythrocyte membrane.

Elasticity↗

PubMed1991

Cytoplasmic rheology of passive neutrophils.

The rheological properties of leukocytes are important to their effectiveness in the microcirculation. Previous studies based on in vitro data from micropipette experiments suggest that a Maxwell fluid bounded by a cortical shell with persistent tension is a realistic model for non-activated neutrophils in both the rapid and slow deformation phases. However, various viscoelastic coefficients have been obtained depending on the degree of cell deformation. In the present paper it is demonstrated that the cytoplasmic apparent viscosity and elasticity vary continuously, depending on the degree of deformation. These apparent variations are due to the inhomogeneous nature of the neutrophil internal structure. It is shown that the nucleus is much stiffer than the cytoplasm. The composite structure of the cell results in the deformation-dependent properties.

Cell Nucleus↗

PubMed1990

Biophysical correlates of lysophosphatidylcholine- and ethanol-mediated shape transformation and hemolysis of human erythrocytes. Membrane viscoelasticity and NMR measurement.

The effects of monopalmitoylphosphatidylcholine (MPPC or lysophosphatidylcholine) and a series of short-chain primary alcohols (ethanol, 1-butanol and 1-hexanol) on cell shape, hemolysis, viscoelastic properties and membrane lipid packing of human red blood cells (RBCs) were studied. For MPPC, the effective membrane concentration to induce the formation of stage 3 echinocytes (8 x 10(6) molecules per cell) was one order of magnitude lower than that needed to induce 50% hemolysis (7 x 10(7) molecules per cell). In contrast, short-chain alcohols induced both shape changes and hemolysis within close concentration range (2.5 x 10(8) to 3.5 x 10(8) molecules per cell). Viscoelastic properties of the RBCs were studied by micropipette aspiration and correlated with shape change. Ethanol-treated RBCs showed a decrease in membrane elastic modulus and an increase in membrane viscosity in the recovery phase at the early stage of shape change. MPPC-treated cells showed the same type of viscoelastic changes, but these were not observed until the formation of stage 2 echinocytes. High-resolution solid-state 13C nuclear magnetic resonance technique was applied to study membrane lipid packing in the ghost membrane by following the chemical shift of hydrocarbon chains. Both MPPC and ethanol caused the 13C-NMR chemical shift to move upfield, indicating that membrane lipids were expanded due to the intercalation of these exogenous molecules. Using data obtained from model compounds, we convert values of chemical shift into a lipid packing parameter, i.e., number of gauche bonds for fatty acyl hydrocarbon chains. Approximately 10(8) interacting molecules per cell are required to induce a detectable change of lipid packing by both MPPC and ethanol. The results indicate that homolysis occurs at a smaller surface area for MPPC- than ethanol-treated RBCs. Our findings suggest that progressive changes in the molecular packing in the membrane lead eventually to hemolysis, but the mode responsible for shape transformation varies with these amphipaths.

Adult↗

PubMed1989

Theoretical and experimental studies on cross-bridge migration during cell disaggregation.

A micromanipulation method is used to determine the adhesive energy density (gamma) between pairs of cytotoxic T cells (F1) and their target cells (JY: HLA-A2-B7-DR4,W6). gamma is defined as the energy per unit area that must be supplied to reduce the region of contact between a conjugated cell pair. Our analysis of the data indicates that the force applied by the micropipette on the cell is not uniformly distributed throughout the contact region as we had previously assumed (Sung, K. L. P., L. A. Sung, M. Crimmins, S. J. Burakoff, and S. Chien. 1986. Science (Wash. DC). 234: 1405-1408), but acts only at the edges of the contact region. We show that gamma is not constant during peeling but increases with decreasing contact area of the conjugated cell pairs F1-JY, F1-F1, and JY-JY in contrast to the constancy of gamma for typical engineering adhesives. This finding supports the notion that the cross-linking protein molecules slide towards the conjugated area across the leading edge of the separation while remaining attached to both cells. Our mathematical analysis shows that the elastic energy stored in the cross-links by the membrane tensions balances the diffusive forces that act against cross-bridge migration. The binding affinity between F1-JY is found to be approximately 15-20 times larger than the corresponding affinity for F1-F1. The number of binding sites of F1 for attachment to JY is approximately the same for binding F1 to another F1 and vary between 10(5) and 10(6).

Animals↗

PubMed1989

Viscoelastic properties of red cell membrane in hereditary elliptocytosis.

The viscoelastic properties of the RBC membrane are in part determined by a submembrane network of proteins consisting of spectrin alpha beta heterodimers (SpD) assembled head-to-head to form spectrin tetramers (SpT) and spectrin oligomers (SpO). SpT, in turn, are connected into a two-dimensional network by the linkage of distal ends of SpT to protein 4.1 and actin. With the micropipette technique, we determined the membrane viscoelastic properties of RBCs from a subset of patients with hereditary elliptocytosis (HE); these RBCs exhibit membrane skeletal instability, defective SpD self-association, and a molecular defect in the alpha I domain of spectrin, which is involved in the SpD-SpD contact (HE SpD alpha-SpD). The elastic modulus and viscosity of the membrane were significantly higher for the HE RBCs than for the control cells. Incubation of normal cells with N-ethyl-maleimide (NEM) produced a similar defective SpD self-association and a significant increase in the viscoelastic parameters of the membrane. The data provide evidence that the mode of assembly of membrane spectrin in the cytoskeletal protein network plays a major role in determining the rheologic behavior of erythrocyte membrane.

Elasticity↗

PubMed1988

Impaired echinocytic transformation of ankyrin- and spectrin-deficient erythrocytes in mice.

The membrane skeleton of the red blood cell plays an important role in the determination of cell deformability and cell shape. Under various in vitro conditions, red blood cells undergo an echinocytic or stomatocytic shape transformation. The mechanism of this fundamental process is not well understood. We have studied the red cell shape transformation in normoblastic anemia mice (nb/nb) and spherocytic anemia mice (sph/sph), which are deficient in ankyrin and spectrin, respectively. We found that both ankyrin-deficient cells (nb/nb) and spectrin-deficient cells (sph/sph) have a reduced capacity to undergo echinocytic transformation with various echinocytogenic treatments, that is, incubation with sodium salicylate (40 and 120 mM), calcium loading (50 microM A23187 + 2.2 mM Ca2+), or metabolic depletion (24 hr at 37 degrees C). These results suggest that the functional integrity of the membrane skeleton is essential for the maintenance and transformation of the red cell shape.

Anemia↗

PubMed1988

Leukocyte relaxation properties.

Study of the mechanical properties of leukocytes is useful to understand their passage through narrow capillaries and interaction with other cells. Leukocytes are known to be viscoelastic and their properties have been established by micropipette aspiration techniques. Here, the recovery of leukocytes to their normal spherical form is studied after prolonged deformation in a pipette which is large enough to permit complete entry of the leukocyte. The recovery history is characterized by the time history of the major diameter (d1) and minor diameter (d2). When the cell is removed from the pipette, it shows initially a small rapid recoil followed by a slower asymptotic recovery to the spherical shape. In the presence of cell activation and formation of pseudopods, the time history for recovery is prolonged compared with passive cell recovery. If a protopod pre-existed during the holding period, the recovery only begins when the protopod starts to retract.

Cell Movement↗

PubMed1988

Passive deformation analysis of human leukocytes.

The following analysis presents an experimental and theoretical study of the passive viscoelastic behavior of human leukocytes. Individual neutrophils in EDTA were observed both during their partial aspiration into a small micropipette and after expulsion from a large micropipette where the cell had been totally aspirated and deformed into a sausage shape. To analyze the data, a passive model of leukocyte rheology has been developed consisting of a cortical shell containing a Maxwell fluid which describes the average properties of the cell cytoplasm. The cortical shell represents a crosslinked actin layer near the surface of the cell and is assumed to be under pre-stressed tension. This model can reproduce the results of experiments using micropipette for both short-time small deformation and slow recovery data after large deformation. In addition, a finite element scheme has been established for the same model which shows close agreement with the analytical solution.

Blood Viscosity↗

PubMed1988

Dynamic changes in viscoelastic properties in cytotoxic T-lymphocyte-mediated killing.

The biophysical properties of cytotoxic T lymphocytes during the killing of their target cells was investigated by using a human cytotoxic T lymphocyte clone, F1, and the target cell, JY, for which it is specific. In single cytotoxic cell/target cell pairs after their conjugation there are changes in the viscoelastic properties of the target cell in association with the lethal hit delivery and post-binding cytolytic steps. On the basis of these changes in the target cell, the complex cytolytic event can be divided into stages: the viscoelastic coefficients exhibited an initial increase followed by a return to resting values; thereafter these coefficients decreased below control and then rose again prior to lysis. The eventual killing of the target cell involves bubbling and swelling of the nucleus, clustering of granules, damage to the cytoplasmic membrane, cell swelling, and lysis. The viscoelastic changes involved in target cell death suggest the loss of integrity of its cytoskeletal apparatus.

Cell Line, Transformed↗

PubMed1987

Rheology of leukocytes.

The rheological properties of human leukocytes (WBCs) have been studied by micropipette aspiration and filtration tests. A small aspiration pressure applied via a micropipette (diameter approximately equal to 3 micron) causes the WBC to undergo a rapid elastic deformation followed by a slow creep. The data can be analyzed with a viscoelastic model: an elastic element K1 in parallel with a Maxwell element (elastic element K2 in series with viscous element mu). Neutrophils and B lymphocytes are similar in K1, K2, and mu, but these values are higher for T lymphocytes. Treatment of neutrophils with colchicine decreases K2 and mu without changing K1, whereas cytochalasin B decreases all three coefficients; these results indicate the importance of cytoskeletal microtubules and microfilaments in WBC rheology. In the presence of Ca2+, WBCs form protopods which have increased viscoelastic coefficients. Inhibition of protopod formation with pentoxifylline is associated with an increase in WBC deformability. The ruffled surface of the apparently round WBC provides an area about twice that needed to enclose a smooth sphere of the same volume; this geometric feature plays an important role in whole WBC deformability tested through 4-5 micron filter pores or micropipettes. Because of its larger volume and higher cellular viscosity, each WBC is equivalent to approximately 700 erythrocytes in its tendency to block 5-micron channels. The rheology of WBCs has significant implications in their functional behavior, including flow through the microcirculation and interaction with endothelial cells.

Capillaries↗

PubMed1987

Rheology of sickle cells and its role in microcirculatory dynamics.

The alterations in the viscoelastic properties of the hemoglobin S solution following deoxygenation can be correlated, on the one hand, with the gelation of the HbS molecules and, on the other hand, with changes in the rheological behavior of the deoxygenated SS erythrocytes. The abnormal rheology of SS erythrocytes forms the basis of their altered flow behavior through filter pores in vitro and microcirculation in vivo. The results support the notion that the rheological behavior of sickle cells is an important link between the molecular events in hemoglobin S following deoxygenation and the behavior of the cell in the microcirculation. Although recent advances have shed considerable lights on the rheology of sickle cells in a variety of conditions, further studies are needed in order to elucidate the pathophysiological role of cell rheology in this disease, especially in terms of the influence of individual SS cells in the heterogeneous populations and the correlation between in vitro and in vivo behavior of SS cells.

Anemia, Sickle Cell↗