The Multi-step Model of

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

(4) Transmigration

Inflamed Tissue
Leukocyte Chemotaxis
in tissue
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 FLOW
(1a) Tethering
HSC
(1b) Rolling
in flow (2) Activation
(3) Firm adhesion

(4) Transmigration
Chemokines

Bone Marrow
HSC Chemotaxis
in tissue
Multistep Model of Leukocyte Transendothelial Migration

von Andrian et al., NEJM, 2000.

 Selectin Expression and Structure
NH 2

*E-selectin3
CD62E
Mature Leukocytes 2,3
and NH 2 *P-selectin
Hematopoietic Progenitors CD62P
COOH

1
*L-selectin
CD62L

NH
2
Endothelial Cell
Cytoplasmic Domain COOH COOH
C-Type Lectin Domain
Transmembrane Domain * Ca++-dependent binding to glycans
1 Constitutive expression, with
EGF-Like Domain 2 shedding
Complement Regulatory Repeat Also expressed on platelets
3
Induced, except in BM and skin
 Parallel Plate Flow Apparatus
Inverted
Vacuum Microscope
Source Video Monitor
w
Flo
Syringe Pump Flow

Cells Flow

CCD
Camera VCR
Outlet Inlet
Cells fixed/grown on
Vacuum 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

Flow Inlet
Outlet Cell
monolayer
Side View Vacuum