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Tuesday, November 24, 2020 | History

2 edition of Leukocyte migration agarose test for the assessment of human neutrophil chemotaxis. found in the catalog.

Leukocyte migration agarose test for the assessment of human neutrophil chemotaxis.

Heikki Repo

Leukocyte migration agarose test for the assessment of human neutrophil chemotaxis.

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Published by Department of Serology and Bacteriology, University of Helsinki in Helsinki .
Written in English


ID Numbers
Open LibraryOL21258535M
ISBN 109514508351

  Neutrophils chemotaxis. Cell migration distances (in mm) are measured microscopically in the direction of the chemoattractant (directed migration (DM)) and towards the control (spontaneous migration (SM)). Neutrophils chemotaxis index is expressed as the ratio of directed to spontaneous migration of PMN cells. x ± SEM; n = 20 subjects per by:   The migration distances were used as indicators of random and directed chemotaxis. After incubation, migration of leukocytes was halted by the addition of 5 mL % gluteraldehyde and mol/L phosphate buffer applied to the center of each plate, allowing the solution to remain on the plate for 30 min (stoppage of leukocyte migration, as. heat-inactivated pooled human serum in the agarose medium because preliminary experiments showed that this substitution did not affect leukocyte chemotaxis. The PMN were suspended in either medium at a concentration of X /ml, and 10 M1 of this suspension was placed in . @article{osti_, title = {Studies of the kinetics of indiumlabeled granulocytes. [Human]}, author = {Weiblen, B J and Forstrom, L and McCullough, J}, abstractNote = {Studies of the in vivo kinetics of granulocytes labeled in vitro with indium were carried out in 10 normal subjects. The granulocyte suspension was prepared with a Ficoll-Hypaque density gradient and labeled with.

To test this, the under-agarose migration assay with freshly isolated human neutrophils was performed (Supplementary Figure S6). H 2 O 2 facilitated neutrophil migration, confirming it as a neutrophil chemoattractant (Figure 7 a). Because benzylamine is an efficient substrate for VAP-1, it was used in experiments with recombinant human VAP-1 Cited by:


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Leukocyte migration agarose test for the assessment of human neutrophil chemotaxis. by Heikki Repo Download PDF EPUB FB2

Optimal conditions for the attraction assay of human neutrophil leukocytes were studied, using the leukocyte migration agarose test (LMAT). Larger migration areas were obtained toward the attractant wells when they were filled with zymosan‐activated serum (ZAS) in advance than when both the attractant and the cells were added by: I.

Effects of environmental factors on neutrophil migration under agarose. Repo H. To apply the leukocyte migration agarose test (LMAT) to the in vitro assessment of human neutrophil chemotaxis, effects of different culture conditions on neutrophil migration under agarose were studied.

Presence of either serum or human serum albumin (HSA) in Cited by: Leukocyte migration agarose test for the assessment of human neutrophil chemotaxis. Variables in the attraction assay. Repo H, Kosunen TU. Optimal conditions for the attraction assay of human neutrophil leukocytes were studied, using the leukocyte migration agarose test (LMAT).Cited by: Leukocyte migration agarose test for the assessment of human neutrophil chemotaxis.

Effects of environmental factors on neutrophil migration under agarose. Repo H. Scand J Immunol, 6(3), 01 Jan Cited by 54 articles | PMID: Leukocyte Migration Agarose Test (LMAT) Assay The mobility of leukocytes plays an important role in immune response to maintain the homeostasis of microenvironment in vivo.

Meanwhile, the leukocyte migration is related to infection and inflammation, which could cause serious complications and interfere to the clinical treatment of many diseases.

Repo H.: Leukocyte migration agarose test for the assessment of human neutrophil chemotaxis. Effects of environmental factors on neutrophil migration under J. Immunol. 6, (). PubMed; Article; CAS; Google ScholarAuthor: J. Sýkora, A. Stajner, V. Palisa, V. Krigar. Abstract.

The technique measuring the migration of PMN neutrophil granulocytes under agarose is simple and inexpensive. Usually results are expressed as the distance migrated by the leading front cells l, but it is also possible to quantitate number of migrating cells 2,3 and observe their morphology y, modifications have been presented for discriminating chemokinesis (enhanced random Cited by: 3.

The under-agarose assay has been used for a variety of purposes, including quantifying chemotaxis (), determining the effects of disease on leukocyte chemotaxis, measuring neutrophil responses to multiple chemotactic gradients (11, 15), and determining the role of various signaling pathways and adhesion molecules in neutrophil by: Repo H, Kosunen TU.

Leukocyte migration agarose test for the assessment of human neutrophil chemotaxis. Variables in the attraction assay. Scand J Immunol. ; 6 (3)– McCall CE, Caves J, Cooper R, DeChatlet L. Functional characteristics of human toxic neutrophils.

J Infect Dis. Jul; (1)– Chemotaxis. Neutrophil chemotaxis was investigated by the under‐agarose method as described previously [12].

Equal volumes of agarose (1%, w/v) and double‐strength RPMI (supplemented with 10% heat‐inactivated FCS) were mixed and allowed to set in 60 mm (diameter) culture dishes, and sets of three wells were by: I. Effect of environmental factors on neutrophil migration under agarose Scand J Immunol 6: () Repo and Kosunen, H.

Repo, T.U. Kosunen, Leukocyte migration agarose test for theassessment of human neutrophil chemotaxis. by: (). Leukocyte migration agarose test for the assessmentof human neutrophil chemotaxis. Variables in the attraction assay. Mechanism of lysosomal enzyme release from human leukocytes.

Microtubule assembly and membrane fusion induced by. Chemotaxis under agarose: a new and simple method for measuring chemotaxis and spontaneous migration of human polymorphnuclear leukocytes and monocytes Nelson, RD; Quie, PG; Simmons, RL Leukocyte migration agarose test for the assessment of human neutrophil chemotaxis.

Increased chemotaxis toward activated serum was demonstrated by leukocytes from bone marrow, spleen, and peripheral blood of tumor-bearing mice as compared with those of normal mice. Three aspects of polymorphonuclear leucocyte (PMN) function were studied in 19 patients with Behçet's syndrome (BS).

By 2 different techniques directed motility was found to be increased. To test the sensitivity of our assay, neutrophil chemotaxis was measured in response to increasing concentrations of IL-8 and LTB4.

A dose-dependent response with concentrations ranging from 0 ng/mL to ng/mL was observed in the presence of IL-8 (Figure 3A).Cited by: 4. The agent of human granulocytic ehrlichiosis (HGE) is an obligate intracellular bacterium with a tropism for neutrophils; however, the mechanisms of bacterial dissemination are not yet understood.

Interleukin-8 (IL-8) is a chemokine that induces neutrophil migration to sites of infection for host defense against pathogens. We now show that HGE bacteria, and the HGE protein, induce IL Cited by: ing in vitro migration under agarose: zymosan activated serum (Zas), containing the chemotactic fragment C5a, the lymphokine eosinophil stimulation promoter (ESP), neutrophil-derived eosinophil chemotactic factor (ECF) and the N-formyl-methionyl-phenylalanine (NFMP) pep­ tide induced chemotaxis and chemokinesis of granulo­ cytes.

Leukocytes navigate through complex chemoattractant arrays, and in so doing, they must migrate from one chemoattractant source to another. By evaluating directional persistence and chemotaxis during neutrophil migration under agarose, we show that cells migrating away from a local chemoattractant, against a gradient, display true chemotaxis to distant agonists, often behaving as if the local Cited by:   To test whether prolonged exposure (30 minutes) to curcumin is a prerequisite to observe this phenomenon, under-agarose migration assay was also performed with untreated neutrophils added to a well cut in agarose polymerized with addition Cited by: Human neutrophils were separated from the blood of normal volunteers by density gradient centrifugation.

In brief, buffy coat cells were obtained from venous blood by dextran T sedimentation, and eosinophils were eliminated by Percoll ( g/ml; Cited by: on neutrophil migration under agarose.

Scand J ImmunolRepo H, Kosunen T U: Leukocyte migration agarose test for the assessment of human neutrophil chemotaxis. Variables in the attraction assay. Scand J Immunol —, John TJ, Sieber OF Jr: Chemotactic migration of neutrophils under agarose Life sci Dose dependence of neutrophil migration under agarose.

Neutrophil responses to a range of IL-8 gradients in the standard 2-h under-agarose migration assay are shown. Neutrophils in the cell well migrate in response to a concentration gradient generated by the Cited by: We found that MIP-2 (at µM) and WKYMVm (at mM) elicited neutrophil intraluminal crawling at similar velocity, neutrophil transendothelial migration and detachment from the venule for comparable length of time, neutrophil migration and chemotaxis in muscle tissue at nearly the same velocity and with similar neutrophil chemotaxis indexes Cited by: 7.

Leukocyte migration from the intravascular space to an eventual inflammatory locus within the lung is a multistep process, dependent on mechanisms directing cell movement to the appropriate site; leukocytes must marginate and adhere to the vascular endothelium, followed by diapedesis between endothelial cells and movement from the vascular.

Neutrophil chemotaxis plays an important role in the inflammatory response and when excessive or persistent may augment tissue damage. The effects of inhibitors indicated the involvement of one or more serine proteinases in human neutrophil migration and.

We hypothesize that LTB 4 is actively secreted by neutrophils as they are migrating toward formyl peptides, therefore acting as a signal-relay molecule. To test this hypothesis, we assessed the role of LTB 4 secretion during primary neutrophil activation and migration in response to formyl peptides.

We find that LTB 4 significantly amplifies neutrophil recruitment to primary chemoattractants Cited by: Assays for the evaluation of inflammatory reactions and chemotaxis by human immune cells were performed in vitro using isolated peripheral leukocytes with the Boyden chamber, 34 agarose gel, and modified agarose gel methods.

35 Depending on the setup and methodological details (eg, stimulus gradient, migration time, depth, filter width, quality Author: Jens Martin Herrmann, Sarah Kirsten Sonnenschein, Sabine Elisabeth Groeger, Nils Ewald, Borros Arnet. "The leukocyte migration agarose test (LMAT) was used to measure quantitatively the magnitude of cell-mediated immunity (CMI) in 35 patients with amoebic liver abscess and 22 healthy controls.

LMAT was positive in % and % of patients with amoebiasis in the presence of µg and µg of the amoeba extract respectively, whereas the test in all 22 healthy controls was by: 2. Neutrophils are fast-moving cells essential for host immune functions. Although they primarily rely on glycolysis for ATP, isolated primary human neutrophils depend on mitochondrial membrane potential for chemotaxis.

However, it is not known whether mitochondria regulate neutrophil motility in vivo, and the underlying molecular mechanisms remain by:   Neutrophil disorders are an uncommon yet important cause of morbidity and mortality in infants and children. This article is an overview of these conditions, with emphasis on clinical recognition, rational investigation, and treatment.

A comprehensive list of references is provided for further by:   The expression of adhesion receptors and diapedesis by polymorphonuclear neutrophils (PMN) were studied before and during experimentally induced Streptococcus uberis mastitis.

Both quarters of the left half of the udders of five midlactation cows were inoculated with a suspension containing approximately CFU of S. uberis J. Clinical signs of an inflammatory reaction and leukocyte Cited by: Chemotaxis has been studied by classical methods that measure chemotactic and random motility responses in vitro, but these methods do not evaluate the total number and phenotype of migrating leukocytes simultaneously.

Our objective was to develop and validate a novel assay, combined Boyden-flow cytometry chemotaxis assay (CBFCA), for simultaneous quantification and phenotypification of.

Patients with septic shock are shown to have decreased neutrophil phagocytic function by multiple assays, and their assessment by whole-blood assays (fluorescence-activated cell sorter analysis) correlates with assays requiring isolated neutrophils (microscopic and spectrophotometric assays).

For patients with similar underlying conditions but without septic shock, this correlation does Cited by: 8. Similarly, neutrophil chemotaxis was decreased ( 6 mm in controls versus 6 mm in sepsis patients [P, ]). No relation between chemotaxis and chemokinesis and other indices of neutrophil function was seen.

The present study confirms previous reports of decreased levels of chemotaxis (2), phagocytosis (20), ROI productionCited by: 8.

Reagents and Antibodies. Eotaxin was obtained from R&D Systems (Abingdon, United Kingdom); platelet-activating factor (PAF) (1-O-hexadecylacetyl-sn-glycerophosphoryl-choline) and formyl-methionyl-leucyl-phenylalanine (fMLP) were from Sigma (St.

Louis, MO); recombinant human interleukin-5 (IL-5) was a gift from M. McKinnon (GlaxoSmithKline, Stevenage, United Kingdom); Ficoll-paque. PKDs contribute to GPCR-controlled chemotaxis of leukocytes. To examine the role of PKDs in leukocyte chemotaxis, we determined the expression profile of PKDs (PKD1, PKD2, and PKD3) in a panel of leukocyte cell lines ().We found that human leukocyte cell line HL60 expressed all three PKD isoforms (Figure 1A and Supplemental Figure S1), and other cell lines expressed one or more isoforms.

Functional Leukocyte Disorders systems,20'21 migration of cells in agarose,22 radiolabeled filter techniques in which the Boyden assay is modified by counting of radiolabeled cells passing Cited by: 2. Neutrophil crawling. (A) Neutrophils demonstrate random migration under non-flow conditions in vitro.(B) Once shear is applied, neutrophils crawl in a coordinated fashion perpendicular to the applied shear.(C) Intraluminal crawling requires an exquisite communication between LFA-1 and MacLFA-1 mediates firm adhesion of neutrophil to the endothelium and would perhaps disengage only Cited by: Cell migration was measured by the leukotactic index (LI); the ratio of migration towards test/migration towards controlResults: Most of the tested dermatophyte concentrations were stimulatory for either neutrophil or monocyte chemotaxis (LI>).

It seemed that osum was the. Effects of PS‐G on neutrophil migration. Neutrophils were preincubated with vehicle or inhibitors ( or n M wortmannin, n M PP2, 10 μ M Ro, 30 μ M PD, or 1–10 μ M SB) for 30 min, and then allowed to migrate for 30 min toward fMLP ( ng ml −1)‐ or PS‐G ( μg ml −1)‐containing medium as indicated Cited by: -Antitrypsin Are Chemotactic for Human Neutrophils A New Paradigm for the Pathogenesis of Emphysema As purified neutrophils are not required for the under agarose migration assays, a mixed leukocyte suspension was used.

Follow- undertaken for each test solution. Neutrophil migration was also independently assessed by.Neutrophil chemotaxis depends on PI3Kγ and Akt/PKB, whereas FMLP fails to induce cell directionality in PI3Kγ-deprived neutrophils.

12 Pharmacologic studies, however, do not indicate an absolute requirement for PIP 3. 13,14 However, not all neutrophil orCited by: