Analysis of individual leucocyte behavior during chemotaxis

Ramsey, W. Scott

doi:10.1016/0014-4827(72)90190-5

Cited by 114 publications

(49 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…However, Zigmond et al (15) did not observe lamellipodial assembly at the site of a prior uropod. The present study and that of Ramsey (27) used buffers in the absence of a gel matrix whereas Zigmond et al (15) used a gel matrix. Indeed, for spontaneous migration in gel matrices, our laboratory has not observed lamellopodium-uropod inversion (4).…”

mentioning

confidence: 94%

See 1 more Smart Citation

Cellular memory: Neutrophil orientation reverses during temporally decreasing chemoattractant concentrations

Albrecht

Petty

1998

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Cell directional orientation or shape polarization is the first cellular step in neutrophil locomotion. To better understand how chemoattractants interact with cells, we studied neutrophil polarization (or shape changes) during exposure to a temporally decreasing chemoattractant signal of N-formyl-methionyl-leucyl-phenylalanine (FMLP) in the absence of a spatial concentration gradient. To accomplish this objective, we used a manifold of differing FMLP concentrations attached to a stopped-f low microscope chamber. Spatial gradients of a f luorescent chemotactic peptide could not be detected in the chamber by using microf luorometry. When FMLP was injected at continually increasing concentrations at 10-s intervals, the shape and relative direction of the neutrophil persisted. However, when temporally decreasing FMLP concentrations were injected, Ϸ80% of the cells changed their direction with 44% of the total cells swinging about to 180°؎ 15°. Most of these directional changes involved dissolution of both the lamellipodium and uropod and reformation of these structures 180°from their original positions. This research suggests that neutrophils reverse their morphological polarity when exposed to temporally decreasing ligand concentrations by ''remembering'' their ligand exposure history and relative direction.Neutrophils participate in inflammatory responses, such as host resistance to infectious disease, and in deleterious host inflammatory reactions, including arthritis, septic shock, and ischemia͞reperfusion injury of tissues during heart attack, stroke, and transplantation. To reach inflammatory sites, neutrophils must first recognize adhesion molecules of endothelial cells lining the circulatory system. Subsequently, neutrophils efficiently cross tissue planes, basement membranes, and interstitial tissues to reach inflammatory foci. Spontaneous neutrophil locomotion involves a series of coordinated cellular processes including oscillatory changes in integrin adhesiveness, signaling, pericellular proteolysis, oxidant production, and actin assembly (1-9). Directed locomotion or chemotaxis additionally involves the ligation of specific chemotaxin receptors, such as the formyl peptide receptors (10). For example, ligation of the formyl peptide receptor leads to the activation of cellular G protein-coupled signaling pathways (11). However, how the activation of G protein pathways promotes chemotaxis and how cells process chemoattractant information during locomotion are uncertain (12, 13).To begin to dissect the problem of neutrophil chemoattractant signal processing, in contrast to simple transmembrane chemical reactions, we have studied cell polarization, the first cellular step in neutrophil locomotion. During polarization for locomotion, neutrophils generate well defined morphological features, including a lamellipodium at the leading edge and a uropod at the trailing end. To focus on the temporal component of signal processing, we used a stopped-flow microscopy chamber to remove the spatial component or...

show abstract

mentioning

confidence: 94%

“…These structures then were reassembled at the opposite sides of the cell, thus forming a 180°change in direction. Ramsey (27) previously has noted this type of behavior. However, Zigmond et al (15) did not observe lamellipodial assembly at the site of a prior uropod.…”

mentioning

confidence: 99%

Cellular memory: Neutrophil orientation reverses during temporally decreasing chemoattractant concentrations

Albrecht

Petty

1998

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…Similar results were obtained using rabbit, human, and guinea pig neutrophils. Since R a m sey [10] showed that cells on a glass surface respond immediately to chemotactic stimulation and since cells which are adhering well also show decreased chemotactic migration when the chamber is turned early, this effect cannot be explained by cell detachment from the starting side of the filter after turning of the chamber. One may assume, therefore, that an ef fect of gravity is superimposed on chemotactic migration in the millipore system, either by influencing the cells or by altering the diffusion rate of the chemotactic material.…”

Section: Om Parison O F Turned An D N On-t Urned C Ham Bersmentioning

confidence: 99%

Assay of Chemotaxis by a Reversible Boyden Chamber Eliminating Cell Detachment

Jungi¹

1975

Int Arch Allergy Immunol

View full text Add to dashboard Cite

A modified in vitro assay for measuring chemotaxis of leucocytes is presented. A closed chamber is used in preference to an open one thus allowing reversal of the chamber during the incubation. The responding cells which at the time the chamber is turned over are inside the filter, then migrate upwards and are consequently retained on the filter by gravity. This chamber is compared with the conventional type and various parameters are examined using guinea pig eosinophils and other cell types. Loss of cells in the conventional Boyden technique is negligible for rabbit peritoneal neutrophils but is considerable for human and guinea pig neutrophils and eosinophils. This is circumvented by the proposed method; accordingly it allows more accurate quantitation in clinical and experimental work.

show abstract

“…(Pediatr Res 25:519-524, 1989) Abbreviations PMN, polymorphonuclear leukocytes NBT, nitroblue tetrazolium fMLP, f-methionylleucylphenylalanine 31D8 MAb, mouse IgGl 31 D8 MAb PMN generally have been assumed to be a homogeneous cell population despite studies as early as the 1920's showing that there were differences among PMN in their ability to move and to ingest bacteria (1)(2). In the past 10 years, numerous studies of PMN structure and function have confirmed that PMN are a heterogeneous group of cells (3)(4)(5)(6)(7)(8)(9)(10). It has previously been shown that PMN from adults differ in their ability to move and that a percentage d o not move at all in response to chemotactic stimuli (1,(3)(4)(5)(6).…”

mentioning

confidence: 99%

“…In the past 10 years, numerous studies of PMN structure and function have confirmed that PMN are a heterogeneous group of cells (3)(4)(5)(6)(7)(8)(9)(10). It has previously been shown that PMN from adults differ in their ability to move and that a percentage d o not move at all in response to chemotactic stimuli (1,(3)(4)(5)(6). Estimates of the size of the poorly motile PMN subpopulation have varied, and the characteristics of this PMN subpopulation have not been well described.…”

mentioning

confidence: 99%

Characterization of Nonmotile Neutrophil Subpopulations in Neonates and Adults

et al. 1989

View full text Add to dashboard Cite

ABSTRACT. Previous studies have demonstrated that polymorphonuclear leukocytes (PMN) are not a homogeneous population of cells but differ significantly in their structure and function. P M N move a t varying rates, and a fraction estimated from 20 to 70% do not move a t all in response to chemotactic stimuli. To characterize this P M N subpopulation better, we studied P M N motility in neonates and adults using a polycarbonate micropore filter chemotactic assay and the 31D8 MAb. Most P M N strongly bind 31D8 MAb (31D8 "bright"), but a minority (31D8 "dull") weakly bind the antibody and in this respect are similar to immature P M N precursors. The 31D8 "dull" P M N have impaired function compared with 31D8 "bright" PMN. In the present study, a P M N subpopulation that failed to migrate using the micropore filter assay accounted for 58 f 7% of adult P M N and was similar to the migrating subpopulation in regard to viability and phagocytic function. The nonmotile subpopulation had a higher percentage of bands (5 f 3% versus I f 2%, p < 0.01) and decreased binding of 31D8 MAb compared with the motile subpopulation. Neonates had a larger nonmigrating P M N subpopulation and 31D8 "dull" P M N subpopulation than those of adults (76 + 3% versus 58 f 7%, p = 0.04 and 26 f 11% versus 8 5 2%, p < 0.01, respectively). These data indicate that although P M N appear morphologically a s a homogeneous population of cells, there exists a viable, nonmotile P M N subpopulation that may be less mature than the motile P M N subpopulation. They also indicate that impaired neonatal P M N motility may be attributable in part to an increased size of the nonmotile P M N subpopulation. of PMN structure and function have confirmed that PMN are a heterogeneous group of cells (3-10). It has previously been shown that PMN from adults differ in their ability to move and that a percentage d o not move at all in response to chemotactic stimuli (1, 3-6). Estimates of the size of the poorly motile PMN subpopulation have varied, and the characteristics of this PMN subpopulation have not been well described. T o further characterize PMN chemotactic subpopulations, we studied PMN motility in healthy adults and in newborn infants using a polycarbonatc filter chemotactic assay and two cell surface MAb (31D8 and Mo-I) binding assays. The 31D8 MAb binds PMN heterogeneously and is associated with PMN motility (9-10). Mo-l binds to a cell surface glycoprotein which helps mediate PMN adherence (I 1 -13). A poorly motile PMN subpopulation was identified in adults and neonates and was found to be larger in neonates. Poorly motile PMN were noted to be viable cells that have similar phagocytic function but decreased binding of 3 1 D8 and Mo-l MAb compared with motile PMN. MATERIALS A N D M E T H O D SStudy popllkulion. Blood was obtained from the fetal side of placentas (neonatal cord blood) within 5 min of birth. Placentas were obtained from healthy women who had delivered vaginally or by repeat cesarian section who had local anesthesia, no complications of pr...

show abstract

Analysis of individual leucocyte behavior during chemotaxis

Cited by 114 publications

References 15 publications

Cellular memory: Neutrophil orientation reverses during temporally decreasing chemoattractant concentrations

Cellular memory: Neutrophil orientation reverses during temporally decreasing chemoattractant concentrations

Assay of Chemotaxis by a Reversible Boyden Chamber Eliminating Cell Detachment

Characterization of Nonmotile Neutrophil Subpopulations in Neonates and Adults

Contact Info

Product

Resources

About