Analysis of CCR7 mediated T cell transfectant migration using a microfluidic gradient generator.
T lymphocyte migration is crucial for adaptive immunity. Manipulation of signaling molecules controlling cell migration combined with in-vitro cell migration analysis provides a powerful research approach. Microfluidic devices, which can precisely configure chemoattractant gradients and allow quantitative single cell analysis, have been increasingly applied to cell migration and chemotaxis studies. However, there are a very limited number of published studies involving microfluidic migration analysis of genetically manipulated immune cells. In this study, we describe a simple microfluidic method for quantitative analysis of T cells expressing transfected chemokine receptors and other cell migration signaling probes. Using this method, we demonstrated chemotaxis of Jurkat transfectants expressing wild-type or C-terminus mutated CCR7 within a gradient of chemokine CCL19, and characterized the difference in transfectant migration mediated by wild-type and mutant CCR7. The EGFP-tagged CCR7 allows identification of CCR7-expressing transfectants in cell migration analysis and microscopy assessment of CCR7 dynamics. Collectively, our study demonstrated the effective use of the microfluidic method for studying CCR7 mediated T cell transfectant migration. We envision this developed method will provide a useful platform to functionally test various signaling mechanisms at the cell migration level.
Department of Physics and Astronomy, University of Manitoba, Winnipeg, MB, R3T 2N2, Canada; Department of Immunology, University of Manitoba, Winnipeg, MB, R3E 0T5, Canada.
Xun
X
Wu
Department of Physics and Astronomy, University of Manitoba, Winnipeg, MB, R3T 2N2, Canada; Department of Biosystems Engineering, University of Manitoba, Winnipeg, MB, R3T 2N2, Canada.
Jiandong
J
Wu
Department of Immunology, University of Manitoba, Winnipeg, MB, R3E 0T5, Canada.
Hongzhao
H
Li
Biotechnology Institute Thurgau (BITg), University of Konstanz, Unterseestrasse 47, CH-8280 Kreuzlingen, Switzerland.
Daniel F
DF
Legler
Department of Immunology, University of Manitoba, Winnipeg, MB, R3E 0T5, Canada.
Aaron J
AJ
Marshall
Department of Physics and Astronomy, University of Manitoba, Winnipeg, MB, R3T 2N2, Canada; Department of Immunology, University of Manitoba, Winnipeg, MB, R3E 0T5, Canada; Department of Biosystems Engineering, University of Manitoba, Winnipeg, MB, R3T 2N2, Canada. Electronic address: flin@physics.umanitoba.ca.
Francis
F
Lin
D002465:Cell Movement / Q000187:drug effects / Q000235:genetics / Q000276:immunology*; D054415:Chemokine CCL19 / Q000276:immunology / Q000494:pharmacology; D002633:Chemotaxis / Q000187:drug effects / Q000235:genetics / Q000276:immunology; D005434:Flow Cytometry; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D019169:Jurkat Cells; D044085:Microfluidics / Q000295:instrumentation / Q000379:methods*; D018613:Microscopy, Confocal; D009154:Mutation / Q000276:immunology; D054400:Receptors, CCR7 / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism; D014162:Transfection
C515370:CCR7 protein, human; D054415:Chemokine CCL19; D054400:Receptors, CCR7; D049452:Green Fluorescent Proteins
CCL19; CCR7; Cell migration; Chemotaxis; Microfluidic device; T cells