Abstract
Transendothelial migration (TEM) of lymphocytes and neutrophils is associated with the ability of their deformable nuclei to displace endothelial cytoskeletal barriers. Lamin A is a key intermediate filament component of the nuclear lamina that is downregulated during granulopoiesis. When elevated, lamin A restricts nuclear squeezing through rigid confinements. To determine if the low lamin A expression by leukocyte nuclei is critical for their exceptional squeezing ability through endothelial barriers, we overexpressed this protein in granulocyte-like differentiated HL-60 cells. A 10-fold higher lamin A expression did not interfere with chemokinetic motility of these granulocytes on immobilized CXCL1. Furthermore, these lamin A high leukocytes exhibited normal chemotaxis toward CXCL1 determined in large pore transwell barriers, but poorly squeezed through 3 μm pores toward identical CXCL1 gradients. Strikingly, however, these leukocytes successfully completed paracellular TEM across inflamed endothelial monolayers under shear flow, albeit with a small delay in nuclear squeezing into their sub-endothelial pseudopodia. In contrast, CXCR2 mediated granulocyte motility through collagen I barriers was dramatically delayed by lamin A overexpression due to a failure of lamin A high nuclei to translocate into the pseudopodia of the granulocytes. Collectively, our data predict that leukocytes maintain a low lamin A content in their nuclear lamina in order to optimize squeezing through extracellular collagen barriers but can tolerate high lamin A content when crossing the highly adaptable barriers presented by the endothelial cytoskeleton.
| Original language | English |
|---|---|
| Pages (from-to) | 239-251 |
| Number of pages | 13 |
| Journal | Journal of Leukocyte Biology |
| Volume | 104 |
| Issue number | 2 |
| DOIs | |
| State | Published - Aug 2018 |
Bibliographical note
Publisher Copyright:©2018 Society for Leukocyte Biology
Funding
We thank Dr. Ann Richmond (Vanderbilt University, Nashville, Tennessee) for providing the CXCR2 expressing HL-60 line. R.A. is the Incumbent of the Linda Jacobs Chair in Immune and Stem Cell Research. R.A. is supported by the Israel Science Foundation, the Flight Attendant Medical Research Institute Foundation (FAMRI), USA, the Minerva Foundation, Germany as well as a research grant from Carol A. Milett. J.L. is supported by awards from the National Institutes of Health [R01 HL082792 and U54 CA210184], the Department of Defense Breast Cancer Research Program [Breakthrough Award BC150580], and the National Science Foundation [CAREER Award CBET-1254846 and MCB-1715606]. We thank Dr. Ann Richmond (Vanderbilt University, Nashville, Tennessee) for providing the CXCR2 expressing HL-60 line. R.A. is the Incumbent of the Linda Jacobs Chair in Immune and Stem Cell Research. R.A. is supported by the Israel Science Foundation, the Flight Attendant Medical Research Institute Foundation (FAMRI), USA, the Minerva Foundation, Germany as well as a research grant from Carol A. Milett. J.L. is supported by awards from the National Institutes of Health [R01 HL082792 and U54 CA210184], the Department of Defense Breast Cancer Research Program [Breakthrough Award BC150580], and the National Science Foundation [CAREER Award CBET-1254846 and MCB-1715606]. S.K.Y. performed most of the experiments, analyzed data, and assisted in manuscript preparation and writing; S.W.F. assisted in FACS analyses and manuscript writing; F.R. performed some of the real time fluorescence microscopy experiments. J.L. provided reagents, expertise, and assisted in the interpretation of results and editing of the manuscript. R.A. designed and supervised experiments and wrote the manuscript. The authors declare no conflict of interest.
| Funders | Funder number |
|---|---|
| Department of Defense Breast Cancer Research Program | BC150580 |
| National Science Foundation | MCB-1715606, CBET-1254846 |
| National Institutes of Health | U54 CA210184 |
| National Heart, Lung, and Blood Institute | R01HL082792 |
| California Breast Cancer Research Program | |
| Flight Attendant Medical Research Institute | |
| Minerva Foundation | |
| Israel Science Foundation |
Keywords
- chemokines
- chemotaxis
- granulocytes
- inflammation
- motility
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