The Multi-step Model of Leukocyte -Endothelial Interactions

Blood
Leukocyte in flow (1a) Tethering (1b) Rolling (2) Activation (3) Firm adhesion

FLOW

(4) Transmigration

Inflamed Tissue
Leukocyte in tissue Chemotaxis

Hematopoietic Cell Engraftment in Bone Marrow

Seeding
Endothelial cells

BM Matrix

“Homing” of HSCs

Bone marrow - identifiable by the fat cells, clusters of developing blood cells, and the large megakaryocytes. Also, in the middle of the field is a cross section of a sinusoid filled with rbc's.

Sinusoid of bone marrow seen here in longitudinal section. There is a nucleus of a lining endothelial cell near the lower center of the field. Junctions between endothelial cells are loose so that HSCs can enter and exit the blood vessels.

Hematopoietic Stem Cell (HSC) Homing to Bone Marrow

Blood
HSC in flow

FLOW
(1a) Tethering (1b) Rolling (2) Activation (3) Firm adhesion

Chemokines

(4) Transmigration

Bone Marrow
HSC in tissue Chemotaxis

Multistep Model of Leukocyte Transendothelial Migration

von Andrian et al., NEJM, 2000.

Selectin Expression and Structure
NH 2

Mature Leukocytes and Hematopoietic Progenitors
COOH

*E-selectin3 CD62E
NH 2

*P-selectin CD62P

2,3

*L-selectin CD62L

1

NH

2

Endothelial Cell
COOH COOH

Cytoplasmic Domain C-Type Lectin Domain Transmembrane Domain EGF-Like Domain Complement Regulatory Repeat

* Ca++-dependent binding to glycans 1 Constitutive expression, with 2 shedding Also expressed on platelets 3 Induced, except in BM and skin

Parallel Plate Flow Apparatus
Vacuum Source Syringe Pump
Flo w

Inverted Microscope Video Monitor
Flow

Cells

Flow

CCD Camera Outlet Vacuum Inlet

VCR
Cells fixed/grown on tissue culture plastic. Protein adsorbed/dried onto glass or plastic.

Flow Chamber

Monolayer Cell Assay

Vacuum Flow Inlet

Top View
Outlet Localization of Rolling Interactions

Outlet

Flow Inlet

Side View

Vacuum

Cell monolayer