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J M Shields

Publications and source records attributed to J M Shields.

29 records · Page 2Linked to original sources

Contraction waves in lymphocyte locomotion.

In this paper we propose that the constriction ring, a prominent feature of moving leucocytes, is a major source of locomotive force. Analysis of time-lapse films of lymphocytes in suspension and moving through three-dimensional collagen gels, demonstrated that the constriction ring was the morphological manifestation of a wave of circular contraction that moved antero-posteriorly. In lymphocytes in suspension the wave moved, although the cells could not. Analysis of lymphocytes moving through a collagen gel revealed that the waves remained stationary with respect to the external environment while the cell appeared to move forward through them. Passage of a single equatorial contraction wave resulted in cell lengthening: a shortening of the region posterior to the constriction was observed in cells moving through collagen gels, but not in lymphocytes held in suspension, suggesting that attachment of cells to the collagen network was necessary for longitudinal contraction. Lymphocyte attachment to collagen gels was mediated through the rapid extension of bleb-like structures into the collagen network. Transmission electron microscopy (TEM) failed to demonstrate any organized structure at the constriction ring. NBD-Phallacidin staining of lymphocytes together with TEM demonstrated that F-actin was distributed evenly throughout the length of the cell. Cell polarity was clearly recognizable by the distribution of coated vesicles, microvilli, and all organelles to the rear, and Thy 1-2 to the front, of motile cells, but polarity could be reversed by the passage of a single contraction wave starting at the rear of the cell, without prior redistribution of these structures.

Actins↗

Invasion of collagen gels by mouse lymphoid cells.

Small mouse lymphocytes from lymph nodes rapidly invaded three-dimensional collagen gels (in the absence of any added chemical attractant). In short-term assays (2-8 hr) this property was restricted to 20-25% of the cell population. Invasion was an active process involving cell locomotion. Time-lapse cinematography revealed that movement was erratic with frequent changes in cell speed. Tracks of cell paths within collagen gels demonstrated that lymphocytes made narrow angles of turn and thus showed a 'persistent random-walk' similar to other cell types moving on plane substrata. Analysis of lymphocyte movement within aligned collagen gels demonstrated that locomotion was biased in the axis of fibre alignment, i.e. lymphocytes showed contact guidance. Separated B lymphocytes invaded collagen gels at a slower rate than unseparated lymph node cells, as also did T cells purified by filtration through nylon wool columns. This latter anomaly implied that nylon wool filtration selectively depleted cells with invasive characteristics from a heterogeneous lymphocyte population. A comparison of Peyer's patch and lymph node lymphocytes showed that both populations invaded at the same rate but the latter cell type did this in greater numbers. This difference may reflect the different proportions of B and T lymphocytes in the two tissues. Lymphocytes from oxazolone-stimulated lymph nodes showed greatly increased movement into collagen matrices compared to unstimulated control lymph node lymphocytes. This increase was demonstrated to be a property of the blast cell population by separating the cells on Percoll gradients into lymphoblast-enriched and -depleted populations.

Animals↗

Separation of mouse lymphoblasts by discontinuous density centrifugation on Percoll gradients.

Mouse lymphoblasts generated in vivo by a topical application of the contact sensitizer oxazolone or by the contents of the gut lumen were separated by discontinuous density centrifugation on Percoll gradients. A 3-step gradient was used to divide the cells into two subpopulations. For cells from oxazolone stimulated lymph nodes, the low density band contained 20-30% of the initial cell number applied to the gradient; 25-40% of this population were in S phase and nearly all the large and pyroninophilic cells were confined to this layer. The high density step cells (70-80% of initial cell number) were predominantly small lymphocytes with less than 0.5% in S phase. Similar results were obtained using cells from picryl chloride stimulated lymph nodes or from mesenteric lymph nodes.

Animals↗

The orientation of fibroblasts and neutrophils on elastic substrata.

The reaction of fibroblasts to the elastic properties of the substratum was studied using elastic collagen films. These films were stretched in one axis to give a substratum which was anisotropic in its elasticity and deformability. Analysis of the orientation of fibroblasts cultured on these substrata showed that they oriented along the axis of stretch which was also the axis of fibre alignment. This orientation was significantly reduced when the films were made less elastic by attachment to a glass slide and chemical fixation. Neither of these procedures appeared to alter the surface shape of these films, which suggests that the elastic properties of the substratum markedly influence the orientation of fibroblasts. The orientation of locomotion of neutrophil leukocytes on elastic collagen films was also analysed and no bias along the axis of stretch was observed. This was compared with neutrophil locomotion in 3-D stretched collagen gels, in which a strong bias along the axis of stretch and of fibre alignment was observed. The possible reasons for the response of these two cell types is discussed.

Animals↗

Lymphocyte locomotion and attachment on two-dimensional surfaces and in three-dimensional matrices.

The adhesion and locomotion of mouse peripheral lymph node lymphocytes on 2-D protein- coated substrata and in 3-D matrices were compared. Lymphocytes did not adhere to, or migrate on, 2-D substrata suck as serum- or fibronectin-coated glass. They did attach to and migrate in hydrated 3-D collagen lattices. When the collagen was dehydrated to form a 2-D surface, lymphocyte attachment to it was reduced. We propose that lymphocytes, which are poorly adhesive, are able to attach to and migrate in 3-D matrices by a nonadhesive mechanism such as the extension and expansion of pseudopodia through gaps in the matrix, which could provide purchase for movement in the absence of discrete intermolecular adhesions. This was supported by studies using serum-coated micropore filters, since lymphocytes attached to and migrated into filters with pore sizes large enough (3 or 8 mum) to allow pseudopod penetration but did not attach to filters made of an identical material (cellulose esters) but of narrow pore size (0.22 or 0.45 mum). Cinematographic studies of lymphocyte locomotion in collagen gels were also consistent with the above hypothesis, since lymphocytes showed a more variable morphology than is typically seen on plane surfaces, with formation of many small pseudopodia expanded to give a marked constriction between the cell and the pseudopod. These extensions often remained fixed with respect to the environment as the lymphocyte moved away from or past them. This suggests that the pseudopodia were inserted into gaps in the gel matrix and acted as anchorage points for locomotion.

Animals↗

Some determinants of the locomotory behaviour of phagocytes and lymphocytes in vitro.

In this review, we discuss some physical and chemical determinants of locomotor behaviour of phagocytic cells and lymphocytes as studied in visual assays, paying particular attention to the following points. (a) A distinction is made between chemokinesis and other forms of kinesis. We propose that the term chemokinesis be reserved for responses resulting from selective recognition of chemical substances. Many kineses in leucocytes may not involve such recognition, but may result from a variety of physical factors, such as those that alter the adhesiveness between cell and substratum. (b) Neutrophils moving in aligned gels of collagen or fibrin show contact guidance of locomotion, i.e. bidirectional movement along the axis of alignment of the fibres of the gel. Thus neutrophils show directional locomotion not only in chemotactic gradients but also in response to physical properties of the substratum. (c) Lymphocytes adhere poorly and move poorly on 2D protein coated substrata. However they move rapidly through 3D collagen gels. This locomotion may be independent of adhesion since lymphocytes may gain traction for locomotion by expanding pseudopods into gaps in the gel matrix, then using the pseudopod as an anchor for subsequent locomotion in any direction.

Animals↗

Relations between Fc receptor function and locomotion in human lymphocytes.

The relationship between the surface binding sites on human lymphocytes for chemotactic factors and for the Fc fraction of IgG was investigated using both blood lymphocytes and established cultures of human lymphoblasts. Pretreatment of human blood lymphocytes with a variety of chemotactic factors inhibited Fc-rosette formation. This was true even of small formylated peptides, for example, formyl-methionyl-phenylalanine (chemotactic) inhibited Fc-rosetting but unformylated methionyl-phenylalanine (non-chemotactic) did not. Conversely pretreatment of lymphocytes with IgG inhibited their locomotor reactions to a variety of chemoattractants. Aggregated IgG was more inhibitory than non-aggregated IgG and the inhibition was mediated via the Fc piece. In a filter assay, native IgG was chemokinetic but not chemotactic for lymphocytes. Heat-aggregated IgG induced more locomotion of lymphocytes than native IgG, and was possibly chemotactic, but no unequivocally so. The possibility that chemotactic factors and the Fc portion of IgG compete for the same cell surface receptor was investigated by binding studies using cultured lymphoblasts. These studies suggested that the reciprocal inhibition could not be explained by competition for receptors. An alternative explanation was suggested by the finding that inhibition of locomotion by aggregated IgG was dependent on the presence of divalent cations at the time the IgG was added, and did not occur in the presence of the calcium ionophore A23187. Addition of aggregated IgG or chemotactic factors to lymphocytes thus may lead to a gated entry of calcium, and following closure of the calcium gate, the cells become relatively unresponsive to further stimulation.

Calcimycin↗

Using pantographic tracings to detect TMJ and muscle dysfunctions.

Forty-six subjects were examined using the HDI and a PRI. Two sets of pantographic tracings were used to determine if subjects experiencing dysfunction could differentiate between degrees of dysfunction. Subjects were divided into groups according to their clinical symptoms: none (D0), sligh (D1), and moderate dysfunction (D5). Each subject was further classified into one of two groups according to his state of occlusion/articulation. The PRI detected differences between the group with moderate dysfunction and those groups with no dysfunction and slight dysfunction on the basis of differences between the first and second sets of tracings; no difference was found between the groups with slight and moderate dysfunction. Subjects with poor occlusions had higher PRI scores. The diagnosis of TMJ dysfunction may require the use of several modalities such as subjective responses, clinical examination, radiographs, and pantographic tracings.

Dental Articulators↗