CD4+ T lymphocytes migrating in three-dimensional collagen lattices lack focal adhesions and utilize beta1 integrin-independent strategies for polarization, interaction with collagen fibers and locomotion.
Cell migration may depend on integrin-mediated adhesion to and deadhesion from extracellular matrix ligands. This concept, however, has not yet been confirmed for T lymphocytes migrating in three-dimensional extracellular matrices. We investigated receptor involvement in T cell migration combining a three-dimensional collagen matrix model with time-lapse videomicroscopy, computer-assisted cell tracking and confocal microscopy. In collagen lattices, the migration of CD4+ T cells (1) involved interactions with collagen fibers at the leading edge and uropod likewise, (2) occurred independently of the co-clustering of beta1, beta2, or beta3 integrins with F-actin, focal adhesion kinase, and phosphotyrosine at interactions with collagen fibers, (3) was counteracted by high-affinity beta1 integrin binding induced by antibody TS2/16; however, (4) the migration could not be blocked by a combination of adhesion-perturbing anti-beta1, -beta2, -beta3, and alpha v integrin antibodies. Integrin blocking neither affected cell polarization, interaction with fibers, beta1 integrin distribution, migration velocity, path structure, nor the number of locomoting cells in spontaneously migrating or concanavalin A-activated cells. Hence, T lymphocytes migrating in three-dimensional collagen matrices may utilize highly transient interactions with collagen fibers of low adhesivity, thereby differing from focal adhesion-dependent migration strategies employed by other cells.
Friedl
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P
Institute of Immunology, University of Witten/Herdecke, Witten, Germany. Peter.fr@mail.uni-wuerzburg.de
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Conrad
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Niggemann
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Zänker
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D000911:Antibodies, Monoclonal / Q000494:pharmacology; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology* / Q000276:immunology / Q000502:physiology*; D002448:Cell Adhesion / Q000276:immunology / Q000502:physiology; D002465:Cell Movement / Q000276:immunology / Q000502:physiology*; D016764:Cell Polarity / Q000276:immunology / Q000502:physiology; D003094:Collagen / Q000378:metabolism; D003208:Concanavalin A / Q000494:pharmacology; D005109:Extracellular Matrix / Q000378:metabolism; D006801:Humans; D007091:Image Processing, Computer-Assisted; D066298:In Vitro Techniques; D019012:Integrin beta1 / Q000502:physiology; D016023:Integrins / Q000502:physiology; D008213:Lymphocyte Activation; D018613:Microscopy, Confocal; D018715:Microscopy, Video
D000911:Antibodies, Monoclonal; D019012:Integrin beta1; D016023:Integrins; D003208:Concanavalin A; D003094:Collagen