F. William Luscinskas


Harvard Medical School
NRB, Room 752P
77 Avenue Louis Pasteur
Boston, MA 2115
Tel: 617-525-4337
Fax: 617-525-4333
Email: fluscinskas@rics.bwh.harvard.edu




The research projects in my laboratory study the mechanisms underlying circulating blood neutrophils (polymorphonuclear leukocyte) and mononuclear (monocyte and T cell subsets) leukocyte adhesion and transmigration into tissues during inflammatory and immune responses 1-4. Defining the molecular mechanisms of leukocyte egress will allow design of rational therapies for treatment of immune and inflammatory diseases.

Our laboratory consists of 5 Research Fellows and a Junior Faculty member who are physician scientists or research scientists and two senior research technicians. who use a combination of immunological, biochemical and molecular biological strategies to study leukocyte recruitment in various in vitro and in vivo models of inflammation. We have developed a valuable in vitro model that allows direct microscopic examination of live leukocyte Ð endothelial interactions under defined laminar fluid shear stress conditions that mimic blood flow in small venules. Areas of focus using this model are three-fold: first, dissection of the adhesion mechanisms that support blood monocyte and specific T cell subset adhesive interactions with endothelium under flow, or specific recombinant endothelial cell adhesion molecules; second, characterization of endothelial-dependent mechanisms involved in regulation of endothelial cell borders (lateral junctions) during leukocyte transmigration, permeability function and cell-cell adhesion; third, using live cell imaging to monitor location of specific leukocyte adhesion molecules (LFA-1, Mac-1, JAM1, PSGL-1) and their counterreceptors on endothelial cell (ICAM-1, JAM-1) during the process of leukocyte arrest, migration and ultimately migration across the endothelium. Biochemical analysis is then used to determine which cytoskeletal linker proteins, such as ezrin or moesin (ERMs family) control the movement of these adhesion molecules and what signaling pathways are involved. My laboratory has taken advantage of the Adenovirus vector gene transfer system 5-7 to introduce Green Fluorescence Protein-labeled wild and mutant molecules at high levels of expression into cultured human endothelial cells to address these questions 8. Specifically, to study lateral junction function during leukocyte transmigration live time under flow, the effort has focused on VE-cadherin (cadherin-5) and Junction Adhesion Molecule (JAM-A,aka JAM1), and claudin-5, which localize selectively to the lateral borders of endothelial monolayers.

Key Words: Inflammation; endothelium; lymphocytes; monocytes; adhesion.

References:

Allport, J. R., H. Ding, W. A. Muller, and F. W. Luscinskas. 2000. Monocytes induce reversible focal changes in VE-cadherin complex during transendothelial migration under flow. J.Cell Biol.203-216.

Grabie, N., M. W. Delfs, Y. C. Lim, J. R. Westrich, F. W. Luscinskas, and A. H. Lichtman. 2002. Beta-galactoside alpha2,3-sialyltransferase-I gene expression during Th2 but not Th1 differentiation: implications for core2-glycan formation on cell surface proteins. Eur.J.Immunol 32:2766-2772.

Ancuta, P., R. Rao, A. Moses, A. Mehle, S. K. Shaw, F. W. Luscinskas, and D. Gabuzda. 2003. Fractalkine preferentially mediates arrest and migration of CD16+ monocytes. J.Exp.Med. 197:1701-1707.

Gerszten, R. E., E. Garcia-Zepeda, Y.-C. Lim, M. Yoshida, H. Ding, M. A. Gimbrone Jr., A. D. Luster, F. W. Luscinskas, and A. Rosenzweig. 1999. MCP-1 and IL-8 trigger firm adhesion of monocytes to vascular endothelium under flow conditions. Nature 398:718-723.

Shaw, S. K., P. S. Bamba, B. N. Perkins, and F. W. Luscinskas. 2001. Real-time imaging of vascular endothelial-cadherin during leukocyte transmigration across endothelium. J.Immunol. 167:2323-2330.



Last Update: 1/6/2014