title: Development and characterization of a physiologically relevant model of lymphocyte migration in chronic lymphocytic leukemia.
abstract: There is growing evidence that lymphocyte trafficking contributes to the clinical course of chronic lymphocytic leukemia (CLL), but to date, only static in vitro cultures have been used to study these phenomena. To address this lack of data, we have developed a dynamic in vitro model in which CLL cells experience shear forces equivalent to those in capillary beds and are made to flow through capillary-like hollow fibers lined with endothelial cells. CLL cells treated in this way increased their expression of CD62L and CXCR4 (both P < .0001) and of CD49d and CD5 (both P = .003) directly as a result of the shear force. Furthermore, CLL cells migrated through the endothelium into the "extravascular" space (mean migration, 1.37% ± 2.14%; n = 21). Migrated CLL cells had significantly higher expression of CD49d (P = .02), matrix metallopeptidase-9 (P = .004), CD38 (P = .009), CD80 (P = .04), and CD69 (P = .04) compared with CLL cells that remained in the circulation. The degree of migration observed strongly correlated with CD49d expression (r(2), 0.47; P = .01), and treatment with the CD49d-blocking antibody natalizumab resulted in significantly decreased migration (P = .01). Taken together, our data provide evidence for a novel, dynamic, and tractable in vitro model of lymphocyte migration and confirm that CD49d is a critical regulator of this process in CLL.
authors.affiliation: Cardiff Chronic Lymphocytic Leukemia Research Group, Institute of Cancer & Genetics, School of Medicine, Cardiff University, Heath Park, Cardiff, United Kingdom;
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Walsby
authors.affiliation: Department of Haematology, King's College London, London, United Kingdom; and.
authors.forename: Andrea
authors.initials: A
authors.lastname: Buggins
authors.affiliation: Department of Haematology, King's College London, London, United Kingdom; and.
authors.forename: Stephen
authors.initials: S
authors.lastname: Devereux
authors.affiliation: Cardiff Chronic Lymphocytic Leukemia Research Group, Institute of Cancer & Genetics, School of Medicine, Cardiff University, Heath Park, Cardiff, United Kingdom;
authors.forename: Ceri
authors.initials: C
authors.lastname: Jones
authors.affiliation: Cancer Research United Kingdom Institute for Cancer Studies, University of Birmingham, Edgbaston, Birmingham, United Kingdom.
authors.forename: Guy
authors.initials: G
authors.lastname: Pratt
authors.affiliation: Cardiff Chronic Lymphocytic Leukemia Research Group, Institute of Cancer & Genetics, School of Medicine, Cardiff University, Heath Park, Cardiff, United Kingdom;
authors.forename: Paul
authors.initials: P
authors.lastname: Brennan
authors.affiliation: Cardiff Chronic Lymphocytic Leukemia Research Group, Institute of Cancer & Genetics, School of Medicine, Cardiff University, Heath Park, Cardiff, United Kingdom;
authors.forename: Chris
authors.initials: C
authors.lastname: Fegan
authors.affiliation: Cardiff Chronic Lymphocytic Leukemia Research Group, Institute of Cancer & Genetics, School of Medicine, Cardiff University, Heath Park, Cardiff, United Kingdom;
authors.forename: Chris
authors.initials: C
authors.lastname: Pepper
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D018929:Cell Culture Techniques / Q000295:instrumentation; D002478:Cells, Cultured; D002634:Chemotaxis, Leukocyte; D018920:Coculture Techniques / Q000295:instrumentation / Q000379:methods; D061307:Human Umbilical Vein Endothelial Cells / Q000502:physiology; D006801:Humans; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000473:pathology*; D008214:Lymphocytes / Q000473:pathology / Q000502:physiology*; D044085:Microfluidics; D008875:Middle Aged; D008962:Models, Theoretical; D033081:Shear Strength