Experimental approaches to lymphocyte migration in dermatology <i>in vitro</i> and <i>in vivo</i>

Radeke, Heinfried H.; Ludwig, Ralf J.; Boehncke, Wolf‐Henning

doi:10.1111/j.0906-6705.2005.00350.x

Cited by 30 publications

(24 citation statements)

References 180 publications

(204 reference statements)

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“…Cell-migration assays were performed in 96-well Transwell plates (Corning, Lowell, MA) with 3.0-mm-pore polycarbonate filters coated with bovine fibronectin (Sigma-Aldrich), essentially as described previously (15,16). Pretreated or nontreated T cells were diluted in RPMI 1640 medium with 0.5% BSA.…”

Section: Chemotaxis Assaysmentioning

confidence: 99%

CXCL9 Causes Heterologous Desensitization of CXCL12-Mediated Memory T Lymphocyte Activation

Giegold

Ogrissek

Richter

et al. 2013

The Journal of Immunology

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The chemokine receptors CXCR3 and CXCR4 are primarily involved in memory Th1 cell–driven autoimmune diseases. Although recent studies in chronic inflammatory disease showed therapeutic success using combined blockade, details of CXCR3 and CXCR4 synergism are not understood. In this investigation, we intended to unravel the interaction of these chemokine receptors in static and dynamic cell-migration assays at both the cellular and molecular levels. Effects of combined stimulation by murine CXCL9 and CXCL12, ligands of CXCR3 and CXCR4, respectively, were analyzed using a murine central memory Th1 cell clone. Costimulation with CXCL9 desensitized the chemotaxis of Th1 cells toward CXCL12 by up to 54%. This effect was found in murine EL-4 cells, as well as in primary human T cells. Furthermore, under dynamic flow conditions CXCL12-induced crawling and endothelial transmigration of Th1 cells was desensitized by CXCL9. Subsequent experiments uncovered several molecular mechanisms underlying the heterologous cross-regulation of CXCR4 signaling by the CXCR3 ligand. CXCR4 surface expression was reduced, whereas CXCL12-induced Akt phosphorylation and intracellular Ca2+ signals were modulated. Moreover, blockade of Rac by NSC23766 revealed differential effects on CXCL12 and CXCL9 chemotaxis and abolished the desensitizing effect of CXCL9. The desensitization of CXCR4 via CXCR3 in memory Th1 cells suggests that their in vivo homeostasis, widely regulated by CXCL12, seemed to be significantly altered by CXCR3 ligands. Our data provide a more detailed understanding for the continuing extravasation and recruitment of Th1 lymphocytes into sites of persistent inflammation.

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Section: Chemotaxis Assaysmentioning

confidence: 99%

CXCL9 Causes Heterologous Desensitization of CXCL12-Mediated Memory T Lymphocyte Activation

Giegold

Ogrissek

Richter

et al. 2013

The Journal of Immunology

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“…Intravital microscopy of mouse ears was performed as described elsewhere. 23,24 Briefly, wild-type-and P-selectin-deficient mice 23 (Jackson Laboratories, Bar Harbor, USA), both backcrossed for more than ten generations on a C57BL6 background, were anesthetized intraperitoneally using ketamin/rompun, and placed on a homeothermic blanket. A catheter was placed micro-surgically into the right carotid artery for injection of CFDA-SE-labeled platelets, efomycine M (4 mg/kg) or Pselectin-specific antibodies (4 mg/kg).…”

Section: Intravital Microscopymentioning

confidence: 99%

Diminished thrombus formation and alleviation of myocardial infarction and reperfusion injury through antibody- or small-molecule-mediated inhibition of selectin-dependent platelet functions

Oostingh

Požgajová²,

Ludwig³

et al. 2007

Haematologica

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BACKGROUND AND OBJECTIVES: P-selectin ctin has been implicated in important platelet functions. However, neither its role in thrombus formation and cardiovascular disorders nor its suitability as a therapeutic target structure is entirely clear. DESIGN AND METHODS: Platelet aggregation was assessed in complementary in vitro settings by measurements of static aggregation, standardized aggregometry and dynamic flow chamber assays. Degradation of aggregates was also analyzed under flow conditions using video microscopy. In vivo, platelet rolling in cutaneous venules was assessed by intravital microscopy in wild-type mice treated with selectin-blocking compounds as well as in P-selectin-deficient mice. FeCl3-induced arterial thrombosis was studied by intravital microscopy in untreated mice or mice treated with an inhibitor of selectin functions. Finally, inhibition of selectin functions was studied in an ischemia/reperfusion injury model in rats. RESULTS: Antibody- or small-molecule-mediated inhibition of P-selectin functions significantly diminished platelet aggregation (p<0.03) and platelet-neutrophil adhesion in vitro (p<0.01) as well as platelet aggregate sizes under flow (p<0.03). Established aggregates were degraded, either via detachment of single platelets following addition of efomycine M, or via detachment of multicellular clumps when P-selectin-directed Fab-fragments were used. In vivo, selectin inhibition resulted in a greater than 50% reduction of platelet rolling in cutaneous venules (p<0.01), producing rolling fractions similar to those observed in P-selectin-deficient mice (p<0.05). Moreover, inhibition of selectin functions significantly decreased the thrombus size in FeCl3-induced arterial thrombosis in mice (p<0.05). In an ischemia/reperfusion injury model in rats, small-molecule-mediated selectin inhibition significantly reduced myocardial infarct size from 18.9% to 9.42% (p<0.001) and reperfusion injury (p<0.001). INTERPRETATION AND CONCLUSIONS: Inhibition of P-selectin functions reduces platelet aggregation and can alleviate platelet-related disorders in disease-relevant preclinical settings

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“…To investigate lymphocyte trafficking into lymphatic tissue of the gut, intravital fluorescence microscopy of Peyer's patches has been used (Miura et al, 1987). Intravital microscopy of the skin and in the dorsal skin fold chamber are attractive models to study leukocyte rolling during inflammation and allergic reactions (Lehr et al, 1993;Radeke et al, 2005). Leukocyte recruitment during ischemia reperfusion injury has been studied by intravital microscopy in several organs such as the liver, intestine, and brain (Liu and Kubes, 2003).…”

Section: Fluorescence Microscopymentioning

confidence: 99%

Analysis of Leukocyte Rolling In Vivo and In Vitro

Sperandio

Pickard²,

Unnikrishnan³

et al. 2006

Methods in Enzymology

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. Novel sulfated lymphocyte homing receptors and their control by a Core1 extension beta 1,3-N-acetylglucosaminyltransferase. Cell 105, 957-969. Yeh, J. C., Ong, E., and Fukuda, M. (1999). Molecular cloning and expression of a novel beta-1, 6-N-acetylglucosaminyltransferase that forms core 2, core 4, and I branches. J. Biol. Chem. 274, 3215-3221. Yousefi, S., Higgins, E., Daoling, Z., Pollex-Kruger, A., Hindsgaul, O., and Dennis, J. W.(1991). Increased UDP-GlcNAc:Gal beta 1-3GaLNAc-R (GlcNAc to GalNAc) beta-1, 6-N-acetylglucosaminyltransferase activity in metastatic murine tumor cell lines. Control of polylactosamine synthesis. J. Biol. Chem. 266, 1772-1782. Zhang, J., Nakayama, J., Ohyama, C., Suzuki, M., Suzuki, A., Fukuda, M., and Fukuda, M. N. (2002). Sialyl Lewis X-dependent lung colonization of B16 melanoma cells through a selectin-like endothelial receptor distinct from E-or P-selectin. Cancer Res. 62, 4194-4198.[23] Analysis of Leukocyte Rolling In Vivo and In VitroBy MARKUS SPERANDIO, JOHN PICKARD, SUNIL UNNIKRISHNAN, SCOTT T. ACTON, and KLAUS LEY Abstract Leukocyte rolling is an important step for the successful recruitment of leukocytes from blood to tissues mediated by a specialized group of glycoproteins termed selectins. Because of the dynamic process of leukocyte rolling, binding of selectins to their respective counter-receptors (selectin ligands) needs to fulfill three major requirements: (1) rapid bond formation, (2) high tensile strength, and (3) fast dissociation rates. These criteria are perfectly met by selectins, which interact with specific carbohydrate determinants on selectin ligands. This chapter describes the theoretical background, technical requirements, and analytical tools needed to quantitatively assess leukocyte rolling in vivo and in vitro. For the in vivo setting, intravital microscopy allows the observation and recording of leukocyte rolling under different physiological and pathological conditions in almost every organ. Real-time and off-line analysis tools help to assess geometric, hemodynamic, and rolling parameters. Under in vitro

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Experimental approaches to lymphocyte migration in dermatology in vitro and in vivo

Cited by 30 publications

References 180 publications

CXCL9 Causes Heterologous Desensitization of CXCL12-Mediated Memory T Lymphocyte Activation

CXCL9 Causes Heterologous Desensitization of CXCL12-Mediated Memory T Lymphocyte Activation

Diminished thrombus formation and alleviation of myocardial infarction and reperfusion injury through antibody- or small-molecule-mediated inhibition of selectin-dependent platelet functions

Analysis of Leukocyte Rolling In Vivo and In Vitro

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