Biomedical subjects

H U Keller

Publications and source records attributed to H U Keller.

At least 19 recordsLinked to original sources

PubMed1991

Analysis of lymphocyte shape by visual classification, calculated measures of shape or light scattering.

Direct visual assays, calculated measures of shape (Dunn and Brown, 1986) and light scattering were applied to detect shape changes and to identify human peripheral blood lymphocytes displaying different shapes (e.g., spherical cells, polarized cells, non-polar cells with surface projections). Spherical cells could be distinguished from polarized cells and from non-polar cells with surface projections by means of calculated measures of shape obtained from outline drawings, the dispersion parameter being more efficient than extension or elongation parameters. Median values for elongation and extension of polarized cells vs. non-polar cells with surface projections differed significantly provided the cell populations consisted entirely of one particular morphological class. However, if the proportion of cells changing shape is small, this may not be detectable by a significant change in the median values. Therefore, we attempted to apply calculated shape factors to identify individual cells showing a particular type of shape change. Spherical cells could be distinguished relatively easily from non-spherical cells on the basis of the dispersion values. However, it was not possible to distinguish unequivocally between polarized cells and non-polar cells with surface projections because the values overlap to a great extent. For this purpose the visual classification was found to be more reliable. Preliminary experiments with light scattering showed that median values for right angle scatter or forward angle scatter are not sensitive enough to permit the detection of a small proportion of cells changing shape.

Humans↗

PubMed1991

Humate-induced activation of human granulocytes.

Naturally occurring humic substances are particular chemical compounds which are found in humus. They bind to carbohydrates, amino acids and steroids by means of hydrogen bonds, covalent bonds and epsilon donor-acceptor complexes. Three specimens of low-molecular humic substances were tested (two naturally occurring humates and one synthetically prepared humate). They were all capable of stimulating certain functions of human neutrophils (PMN), such as the respiratory burst which results in the production of toxic oxygen compounds. This PMN stimulation can be demonstrated with the help of chemiluminescence, as well as by cytochemistry and with the electron microscope. The main product of the humate-induced PMN response is H2O2. There was no activation of neutrophilic chemokinesis or chemotaxis. It is suggested that the low-molecular humic substances originating from decaying organic material contain chemical structures which can act as signals to change dormant PMN into activated cells.

Acridines↗

PubMed1990

Diacylglycerols and PMA are particularly effective stimulators of fluid pinocytosis in human neutrophils.

Diacylglycerols (OAG, diC8) and PMA were found to stimulate fluid pinocytosis (net uptake of FITC-dextran) to a far greater extent than other neutrophil activators, such as the chemotactic agents fNLPNTL and LTB4, the microtubule disassembling agents colchicine and nocodazole, the kinase inhibitor H-7, or D2O. OAG and diC8 produce a dose-dependent increase in the uptake of FITC-dextran, which is up to about 25- to 30-fold the control value of unstimulated neutrophils. The protein kinase inhibitor H-7 alone had a small stimulating effect on the net uptake, and it failed to inhibit stimulation of fluid pinocytosis by PMA, OAG, and diC8. Also, the protein kinase inhibitor staurosporine failed to inhibit fluid pinocytosis stimulated by OAG, diC8, and PMA. Stimulated fluid pinocytosis and vacuolization in response to PMA or diacylglycerols is associated with surface ruffling of neutrophils. Pinocytosis as well as surface ruffling stimulated by PMA, OAG, diC8, or diC10 are suppressed in the presence of cytochalasin D. The results suggest that diacylglycerols may be instrumental in transducing the signal for stimulated pinocytosis and that the surface movements induced by diacylglycerols, and PMA may be instrumental in fluid pinocytosis.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗

PubMed1990

Induction of monocyte chemotaxis in devascularized rabbit bone.

Temporary impairment of blood supply has been suggested to cause bone remodeling. The degradation of cells and matrix and the attraction of resorbing cells were examined in this study. Bone specimens of rabbits were stored in vitro for 2-20 days. At the end of this aging process the probes were tested for their chemotactic activity toward autologous leukocytes in a diffusion chamber. Both supernatant from the aged bone specimens and ground bone particles exhibited significant chemotactic activity that was specifically attracting monocytes. It is suggested that soluble bone matrix proteins or degeneration products liberated during ischemic damage to cortical bone initiate the resorptive process.

Animals↗

PubMed1990

Expression of cellular effector functions and production of reactive nitrogen intermediates: a comparative study including T lymphocytes, T-like cells, neutrophil granulocytes, and mononuclear phagocytes.

The ability of various cell types to secrete reactive nitrogen intermediates (RNI) upon functional activation was comparatively assessed. Neither in T lymphocyte clones mediating MHC class I or class II antigen-restricted killing via alpha/beta T cell receptor (TcR) or MHC-unrestricted killing via gamma/delta TcR, nor in peripheral blood mononuclear cells expressing natural killer or lymphokine-activated killer activity, target cell lysis was associated with detectable RNI production. Also, activated neutrophil granulocytes did not secrete RNI. In contrast, bone marrow-derived mononuclear phagocytes, activated to express tumoricidal activity, secreted marked RNI activity.

Animals↗

PubMed1990

The protein kinase C inhibitor H-7 activates human neutrophils: effect on shape, actin polymerization, fluid pinocytosis and locomotion.

The present study demonstrates new properties of H-7. The protein kinase inhibitor H-7 is a potent activator of several neutrophil functions. Stimulation of initially spherical nonmotile neutrophils elicits vigorous shape changes within a few seconds, increases in cytoskeletal actin, altered F-actin distribution, increased adhesiveness and a relatively small increase in pinocytic activity. H-7 has also chemokinetic activities. Depending on the experimental condition, H-7 may elicit or inhibit neutrophil locomotion. It failed to induce chemotaxis. Thus, the response pattern elicited by H-7 is different from that of other leukocyte activators such as chemotactic peptides, PMA or diacylglycerols. The finding that H-7 can elicit shape changes, actin polymerization and pinocytosis suggests that these events can occur without activation of protein kinase C (PKC). PMA-induced shape changes and stimulation of pinocytosis were not inhibited by H-7.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗

PubMed1989

Diacylglycerols and the protein kinase inhibitor H-7 suppress cell polarity and locomotion of Walker 256 carcinosarcoma cells.

We show that diacylglycerols, like phorbol myristate acetate (PMA), suppress cell polarity and locomotor activity of Walker carcinosarcoma cells in a dose-dependent fashion in vitro. OAG and diC8 show significant activity at concentrations above 3 x 10(-5) M. The inhibitory effect on locomotion is due to a reduction in the proportion of locomoting cells rather than gradual lowering of the speed of individual cells. Measurement of protein kinase C (PKC) activity in isolated fractions showed a substantial reduction of the total cellular PKC activity and of the activity in the cytosolic fraction following incubation of cells with 10(-8) M PMA for 30 min. In contrast, the total and relative PKC activity associated with the membrane fraction was increased by PMA. The effect of H-7, an inhibitor of PKC as well as of cAMP-dependent kinase, has been tested. H-7 suppressed cell polarity of "unstimulated" control cells (ID50 = 6.5 microM H-7), colchicine-stimulated cells (ID50 = 92 microM H-7) or cells treated with both PMA and colchicine (ID50 = 15 microM H-7), in a dose-dependent fashion. The locomotor activity of the cells was also suppressed. LTB4 had no clearcut activity in this system. Our findings suggest that diacylglycerols and H-7 are of interest as physiological or pharmacological stop signals for tumor-cell locomotion. Contrary to our expectations, PMA and diacylglycerols vs. H-7 did not produce opposing or antagonistic effects on cell polarity and locomotion. This similarity may be due to down-regulation of PKC by PMA and inhibition of PKC by H-7. However, the mechanisms underlying these novel effects of diacylglycerols and of H-7 on cell polarity and locomotion may be even more complex; they require further studies.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗

PubMed1989

Diacylglycerols and PMA induce actin polymerization and distinct shape changes in lymphocytes: relation to fluid pinocytosis and locomotion.

Shape changes have been determined in human blood lymphocytes stimulated with OAG, diC8, PMA, colchicine or the hexapeptide fNLPNTL in short-term assays (30 min). Distinct types of shape-change responses were observed. Colchicine was active in generating a relatively small proportion of polarized lymphocytes (front-tail polarity). OAG, diC8 and PMA produced different types of shape change (non-polar cells with surface projections), and these were closely associated with an increase in actin polymerization and a shift of F-actin into the projections at the cell periphery. The diacylglycerols OAG and diC8 produced biphasic dose-response curves leading to rounding up of cells at very high stimulant concentrations. PMA produced no comparable biphasic response when tested over a much wider concentration range. Though the nonpolar cells with surface projections generated by OAG, diC8 or PMA showed vigorous shape changes, they lacked significant locomotor activity. alpha-Phorbol, 4 alpha-PDD, lumicolchicine or fNLPNTL were inactive. Small blood lymphocytes stimulated by OAG, diC8 or PMA showed a very small increase in the net uptake of FITC-dextran by fluid pinocytosis. Unlike neutrophils, which show a high net uptake, lymphocytes did not concentrate FITC-dextran in large granules, indicating that they do not develop a 'storage' compartment in the form of large vesicles. However, small fluorescent spots were consistently found in at least a fraction of blood lymphocytes. The results indicate that stimulated surface movement may be instrumental in fluid pinocytosis. Diacylglycerols may act as second messengers to induce pinocytosis, shape changes and altered actin polymerization in lymphocytes.

Actins↗

PubMed1988

Diacylglycerol-induced shape changes, movements and altered F-actin distribution in human neutrophils.

The study shows that diacylglycerols (DAGs) as physiological activators of protein kinase C induce characteristic shape changes in human neutrophil granulocytes. In contrast to chemotactic peptides, which can induce front-tail polarity characterized by a contracted tail and an expanding front, DAGs elicit the formation of non-polar cells with surface projections. These cells exhibit a distinct type of motility characterized by vigorous and continuous shape changes without front-tail polarity and without the unidirectional movement and cytoplasmic streaming seen in polarized cells. In neutrophils exposed to DAGs, F-actin is shifted to the cell periphery and mainly into the surface projections of activated cells. DAGs induce the formation of large intracellular vacuoles in neutrophils producing surface projections, and these vacuoles persist after the cells have reacquired a spherical shape. Combined stimulation of human neutrophils with DAG and fNLPNTL results in a suppression of peptide-induced polarity and the formation of non-polar motile cells resembling those stimulated with DAG alone. These results suggest that the diacylglycerol-protein kinase C pathway may be instrumental in transducing or modulating signals to both the locomotor apparatus and the exocytotic and/or pinocytic system of the cell. Neutrophil stimulation with DAGs thus represents a useful model with which to study further the hypothesis that distinct types of neutrophil shapes and movements are preferentially associated with distinct functions and to characterize signalling pathways.

Actins↗

PubMed1988

Heavy water (D2O)-induced shape changes, movements and F-actin redistribution in human neutrophil granulocytes.

Heavy water (D2O) induces characteristic shape changes and a distinct type of movement in human neutrophil granulocytes. In contrast to front-tail polarity as evoked by chemotactic peptides and microtubule-disassembling agents, D2O-based media produce non-polar neutrophils with many small or long surface projections. This phenotype is similar to that elicited by both phorbol myristate acetate and diacylglycerols, but the surface projections are smaller and more densely placed and are often associated with a single large projection. D2O-induced non-polar cells with surface projections perform continuous shape changes without front-tail polarity and without the unidirectional movement and cytoplasmic streaming seen in cells with front-tail polarity. Some of the cells show circus movements of a large projection indicating circular polarity. In neutrophils suspended in D2O, F-actin is shifted to the cell periphery, mainly into the surface projections of activated cells. The D2O-induced effects are reversed in H2O-based medium. D2O is dominant over the chemotactic peptide, N-formyl-L-methionyl-L-leucyl-L-phenylalanine (fMLP), colchicine and taxol in that the combined action of D2O with any of these agents results in the D2O-induced phenotype. In contrast, cytochalasin B alone and in combination with fMLP induces a considerable decrease of non-polar cells and an increase of spherical cells similar to non-stimulated cells in H2O-based medium. Earlier studies indicated that D2O acts on microtubules. Our results suggest that D2O may act on the microfilament system. Neutrophils suspended in D2O-based medium may represent a useful model to study the relationship between shapes, movements, and particular functions of these cells.

Actins↗

PubMed1987

Effect of phorbol myristate acetate and the chemotactic peptide fNLPNTL on shape and movement of human neutrophils.

The results show that the distinct types of shape produced by phorbol myristate acetate (PMA) and by chemotactic peptides (fNLPNTL) are associated with distinct types of neutrophil movement. Whereas the chemotactic peptide can induce front-tail polarity characterized by an expanding front, a contracted tail and preferential unidirectional movements of intracellular organelles, PMA can only elicit non-polar movements characterized by random formation and retraction of projections all over the surface, intracellular movements of organelles being ill-defined and changing in direction. Combined stimulation of human neutrophils with PMA and fNLPNTL results in a suppression of peptide-induced polarity and the formation of non-polar motile cells resembling those stimulated with PMA alone. The results suggest that the diacylglycerol-protein kinase C pathway may be instrumental in transducing or modulating signals to the locomotor apparatus of the cell. PMA-treated cells are, however, still capable of developing directional responses when appropriately stimulated. The findings lead to the hypothesis that distinct types of neutrophil movements are preferentially associated with distinct functions.

Chemotaxis, Leukocyte↗

PubMed1987

Shape, movement and function of neutrophil granulocytes.

Evidence that distinct types of shapes and movement can be induced by different neutrophil-activating agents like the chemotactic peptides and PMA is reviewed. Front-tail polarity seems to be preferentially associated with locomotion, whereas non-polar cells with surface projections show preferentially pinocytosis. The following partially hypothetical concept on neutrophil shape changes has been derived from these experiments: Neutrophils are capable to perform distinct types of movements depending on the stimulus or stimuli. Distinct movements are preferentially associated with distinct functions such as locomotion, pinocytosis or others. The underlying cytoskeletal changes may differ accordingly.

Animals↗

PubMed1987

Locomotion of tumor cells as an element of invasion and metastasis.

Malignant tumor cells are endowed with the ability to invade host tissues and to produce metastases. In this review, tumor cell locomotion as an important pathogenic mechanism in the invasive process is discussed. The traffic of neoplastic cells along preformed or newly created tissue pathways will not only depend on cellular cues and/or extracellular stimulatory or inhibitory soluble factors, but also on the interactions of tumor cells with the invaded tissue. Normal tissue exerts its influence on tumor cell migration mainly by providing contact surfaces, as both the growth and locomotor phenomena of tumor cells depend on the cells ability to adhere to structures present along the invasion front. Migrating tumor cells are morphologically characterized by a cell shape change typical for locomoting cells, i.e. polarization. This locomotor phenotype is associated with altered functions of the cytoskeleton induced by agents acting on cell surface receptors, on the signal cascade, or on the cytoskeletal apparatus itself, or evoked by mutations of cytoskeletal proteins. The search for stop signals of tumor cell locomotion is of particular interest, as this may represent an approach to exert an influence on the invasive process.

Animals↗

PubMed1986

Shape changes and chemokinesis of Walker 256 carcinosarcoma cells in response to colchicine, vinblastine, nocodazole and taxol.

Two morphologically distinct types of polarization and locomotion are found in Walker carcinosarcoma cells. Spontaneously polarized cells are characterized by formation of ruffles and spikes, cells polarized in the presence of microtubule-disassembling drugs by blebbing at the leading edge. Polarization and locomotion are closely correlated. Both can be stimulated by the microtubule-disassembling agents colchicine, nocodazole and vinblastine and suppressed by taxol or cytochalasin B. The results suggest that both microfilaments and microtubules may be involved in regulating polarization and chemokinesis, and that shape changes elicited by microtubule-disassembling agents depend on microfilaments.

Alkaloids↗

PubMed1986

Qualitative and quantitative assessment of human polymorphonuclear leukocyte (PMNL) functions.

The current status on human PMNL separation, storage conditions, and quantitative assessment of cell functions is discussed. Human peripheral blood PMNLs are multifunctional cells which may be activated by several soluble and particulate stimuli. A variety of methods for the isolation of PMNLs and testing of cell functions is described which may be used in the clinical and the research laboratory. Detailed information on the quantitative assessment of PMNL motility, phagocytosis, microbial killing, exocytosis, and oxygen metabolite generation is given. Recently developed biochemical and immunological methods which may be employed for detection of structural and metabolic changes in normal and pathological PMNLs are also included.

Blood Bactericidal Activity↗

PubMed1985

Phorbol myristate acetate (PMA) suppresses polarization and locomotion and alters F-actin content of Walker carcinosarcoma cells.

This study demonstrates a novel feature of PMA, its ability to suppress chemokinetic polarization and locomotion of tumor cells. Walker carcinosarcoma cells exhibit two distinct types of polarization and locomotion, i.e. spontaneous polarization characterized by ruffles at the front and stimulated polarization and locomotion in response to the microtubule-disassembling agents colchicine, vinblastine and nocodazole, which are characterized by blebbing at the front. The tumor promotor phorbol myristate acetate (PMA), but not phorbol, was found to suppress both types of polarization and random locomotion at concentrations between 10(-8) and 10(-6)M. The effect of 10(-6)M PMA was virtually complete within 5 min. Inhibition of locomotion was due to both a reduction in the speed of migrating cells and the proportion of migrating cells. Changes in shape and chemokinesis of Walker carcinosarcoma cells were associated with alterations in the relative amount and the topographical distribution of F-actin as determined by NBD-phallacidin binding. Suppression by PMA was associated with loss of the polar topographical distribution of F-actin visualized by NBD-phallacidin binding. In the presence of PMA, the relative amount of F-actin was higher than in unstimulated controls and lower in cells exposed to microtubule-disassembling agents.

Actin Cytoskeleton↗

PubMed1985

Orthokinetic and klinokinetic responses of human polymorphonuclear leucocytes.

Evidence is presented to show that klinokinesis, which was previously demonstrated in bacteria and amoeba only, may also occur in metazoan cells. The chemotactic peptide formyl-Met-Leu-Phe (fMLP) elicited orthokinetic and klinokinetic responses of human blood-borne polymorphonuclear leucocytes (PMNs) under the test conditions used. Increased speed (orthokinesis) was due to an increase in the proportion of migrating cells as well as in the speed of the locomoting subset. The klinokinetic effect was manifested by a decrease in the klinolocomotion index, the mean angle of changes in direction greater than or equal to 90 degrees, and the frequency of turns greater than or equal to 90 degrees. The klinolocomotion index was inversely related to speed. This explains the synergistic effect of klinokinesis and orthokinesis in this system. Colchicine alone had an orthokinetic effect which was exclusively due to alterations in the proportion of migrating cells and it altered the turning behaviour without exerting a klinokinetic effect. However, colchicine had marginal orthokinetic and klinokinetic effects on fMLP-stimulated cells resulting in reduced translocation. The relationship between klinokinesis and mean angle or frequency of turns has been analysed. Klinokinesis was a substantial though not the major element of the chemokinetic response to fMLP under the conditions used. No other metazoan cells have been shown to possess such a complete pattern of responses, including orthokinesis, klinokinesis, and chemotaxis, which regulate locomotion.

Bacterial Physiological Phenomena↗

PubMed1979

Chemotaxis is not a special case of haptotaxis.

Serum peptides containing classical anaphylatoxin (CAT) produce marked chemotatic orientation of human neutrophil granulocytes without modifying cell attachment to the substratum. Furthermore gradients of adhesion produced with gammaglobulins fail to induce morphological orientation of neutrophils. The results suggest that chemotaxis is not a special case of haptotaxis.

Anaphylatoxins↗