Biomaterials as Adjuvants

Last Time: drug targeting

Today: Delivering activation signals to dendritic cells in vaccines

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ANNOUNCEMENTS:

Lecture 24 Spring 2006 1

 Targeting vaccines to dendritic cells
Targets on DCs:
DEC-205
CD11c
TLR3
…particles, in general

Images removed due to copyright restrictions.

Please see: Kwon, et al. PNAS 102 (2005): 18264-18268.

Lecture 24 Spring 2006 2

 Figure removed due to copyright restrictions.
Please see: Figure 4 in Kwon, et al. PNAS 102 (2005): 18264-18268.

Lecture 24 Spring 2006 3

‘REVERSE TARGETING’, CONTINUED

Lecture 24 Spring 2006 4

 Targeting dendritic cells to
vaccines: ‘Reverse targeting’ to Infection site
mimic infection site recruitment
2) Antigen
loading and
1) Attraction to sites of infection
activation

Infected cells 3) Trafficking

to lymph
nodes

4) Activation of
naïve T cells in
the lymph nodes
1) Chemotaxis:
Migration ‘up’ concentration
gradients of chemoattractantLecture 24 Spring 2006 5
 O CH3 O
H2
PLGA C CH O
C
C
O
C
C
O
CH O C
x H2 y
CH3 O O

‘Vaccination
site’ source well
~1 mm

1.2 mm
3 mm
collagen gel

Images removed due to copyright restrictions.

Please see: Zhao, X., et al. Biomaterials 26 (2005): 5048.

Lecture 24 Spring 2006 6

 Dendritic cell attraction, antigen loading,
and activation

Alginate
microsphere
Antigen-delivery
nanoparticles

DCs

chemokine

Lecture 24 Spring 2006 7

How to encapsulate multiple
factors under mild conditions
for ‘reverse targeting’? ALGINATE
COOH COOH H H
O O O O
H H H H
H H COOH COOH
O O O O
OH OH OH OH OH OH OH OH
H H H H

H H H H H H H H

Mannuronic acid block Guluronic acid block

Figure by MIT OCW.
Image removed due to copyright restrictions.
alginate Please see: http://www.lsbu.ac.uk/water/hyalg.html
CaCl2 solution

Wash

Iso-octane
Lecture 24 Spring 2006 8
 Fluorescent nanoparticles Fluorescent chemokine

100

80

% MIP-3α Released
60

40

20

0
0 1 2 3 4 5 6 7

Lecture 24 Spring 2006 Time (Days) 9

 200 200

y - d is ta n c e fro m b e a d

y- distance from bead

100 100

0 0
-200 -100 0 100 200 -200 -100 0 100 200

-100 -100

collagen gel
-200 -200
x- distance from bead x- distance from bead

25

Migration Histogram N=100

20

15

# Cells
10

0
25

75

5
12

17

22

27

32

37

42
Distance (μm)

Lecture 24 Spring 2006 10

 Alginate microspheres loaded with:

Antigen nanoparticles Antigen nanoparticles

+ MIP-3α
# cells

3.9% 17.9%

Amount of DQ-ovalbumin internalized

Lecture 24 Spring 2006 11
 Issues in targeted delivery

• Where is the target molecule expressed?

• Is it expressed by normal tissues?
• Is it stably expressed?
• Can select out evasive tumor cells/viruses

• What is the affinity of binding?

• immune response to targeting agent

Lecture 24 Spring 2006 12

 ADJUVANTING VACCINES WITH
SYNTHETIC MATERIALS
MIMICKING PATHOGEN-HOST INTERACTIONS TO STIMULATE
IMMUNITY

Lecture 24 Spring 2006 13

 ADJUVANTING FUNCTIONS OF BIOMATERIALS

• Sustain delivery of antigen

– Extracellular or intracellular

• Mimic pathogen delivery of activation

signals to dendritic cells and B cells
– Mimic multivalent surface structure of
pathogens
• Limit dose, but enhance response

Lecture 24 Spring 2006 14

 SUSTAINING ANTIGEN DELIVERY TO DENDRITIC CELLS

EXTRACELLULAR DEPOTS INTRACELLULAR DEPOTS

Images removed due to copyright restrictions.

Please see: Shen, et al. Immunology 117 (2005): 78-88.

Lecture 24 Spring 2006 15

 pathogens as biomaterials: how the structure of pathogens relates
to immune responses

Image removed due to copyright restrictions.

Please see: http://www.kripafoundation.org/ Rehabilitation%20Programs/faqhiv.html

Image removed due to copyright restrictions.

Please see: http://textbookofbacteriology.net/BSRP.html

Lecture 24 Spring 2006 16

pathogens as biomaterials: how the structure of pathogens relates
to immune responses

Lecture 24 Spring 2006 17

 Dose sparing of adjuvants by co-delivery in particles
Many immunostimulatory factors that activate
DCs are also potent inflammatory stimuli:

T cell proliferation measured ex vivo 10d after 2 injections:

Image removed due to copyright restrictions.

Please see: Heikenwalder, et al. Nat Med 10 (2004): 187-192.

Image removed due to copyright restrictions.

Please see: Diwan, et al. Current Drug Deliv 1, no. 4 (2004): 405-412.

Lecture 24 Spring 2006 18

 Nanoparticles that mimic pathogen
structural features

(1) Particulate
antigen delivery (2) surface display
- - -+
- - of repeated native

-
+
-

-+
+

-
+

Ag epitopes
+
+

-
+
-+

- -
-
+ +
+ +
+
- (3) surface display
of ‘pathogen
+

-
- -

+
+

-
+ associated
+ -
-

-
-
+

500 nm CpG oligos molecular patterns’

Poly(L-arginine)

Lecture 24 Spring 2006 19

 Image removed due to copyright restrictions.
Please see: Jain, S., et al. Biomacromolecules 6 (2005): 2590.

Lecture 24 Spring 2006 20

 Images removed due to copyright restrictions.
Please see: Jain, S., et al. Biomacromolecules 6 (2005): 2590.

Lecture 24 Spring 2006 21

Proposed mechanism for antigen delivery to dendritic cells

Soluble Ova + CatD

1.

3.

2.

Graphs removed due to copyright restrictions.

Please see: Jain, S. et al. Biomacromolecules 6 (2005): 2590.

Lecture 24 Spring 2006 22

 Cytokine Secretion by Activated DCs

18000
160 soluble CpG
CpG
15000 particles
particles
particle-CpG particle-CpG
120
IL-12p70 (pg/ml)

12000

80 IL-6 (pg/ml) 9000

6000
40
3000

0 0
0.001 0.01 0.1 1
0.001 0.01 0.1 1
CpG concentration (PM) CpG concentration (PM)
Lecture 24 Spring 2006 23
 pathogens as biomaterials: how the structure of pathogens relates
to immune responses

Image removed due to copyright restrictions.

Please see: http://www.kripafoundation.org/ Rehabilitation%20Programs/faqhiv.html

Image removed due to copyright restrictions.

Please see: http://textbookofbacteriology.net/BSRP.html

Lecture 24 Spring 2006 24

 B Cell Activation In Vivo

week 1 week 3 week 5 week 7

70000

60000

50000
Anti-ova IgG titers

40000

30000

20000

10000

0
ova ova + CpG particles particles + PpR60-CpG ova + CFA
CpG
Samples

Lecture 24 Spring 2006 25

 Synthetic pathogens?

Image removed due to copyright restrictions.

Please see: Troutier, et al. Langmuir 21 (2005): 1305-1313.

I II III

SAv biotin-ova
biotinylated
phagocytosis
ligand mobile ligand particles

Lecture 24 Spring 2006 26

 Synthetic pathogens?

Figures removed due to copyright restrictions.

Please see: Figures 3 and 5 in Yu, et al. Adv Mater 17 (2005): 1477-1480.

Lecture 24 Spring 2006 27