Download Bryan 2012 Research Day Poster

Characterization of Pulmonary Microvesicles
Bryan Prahl and Dr. James Burritt
University of Wisconsin-Stout Menomonie, Wisconsin
Methods:
Fig. 1: Microscopy of
pulmonary microvesicles using
a Zeiss Axioscope 2-Plus
microscope.
Fig. 2: Measure of superoxide
production using a Veritas
luminometer .
Fig. 3: Protein analysis of
pulmonary microvesicle
samples at different time points
from BALF of C57 mice, using
a 4-15% Tris-HCl gel
electrophoresis.
X, XO
A
1500000
X, XO
X
X, XO, SOD
MCLA Rgt
1000000
RLU
•Pulmonary exosomes have
reported to regulate the
immune system through
several mechanisms.
•Pulmonary microvesicles are
similar to exosomes, but twenty
times larger on average.
•Microvesicles are unreported
to the best of our knowledge.
•Our data suggest pulmonary
microvesicles do not produce
antimicrobial reactive oxygen
species, but does upregulate a
specific protein when exposed
to a pathogen such as
Aspergillus fumigatus.
•We are collecting further data
to better describe these
unreported pulmonary vesicles.
Proteins Associated
with Microvesicles
Superoxide
Production
Morphology of
Microvesicles
500000
0
0
10
20
30
Time (min)
B
BALF - MCLA
75000
MV 2 Hours
MV 7 Hours
MCLA
50000
RLU
Overview:
25000
A A
A
250
150
100
75
B C D E F
50
37
25
20
15
10
0
0
25
50
75
100
Summary
•Pulmonary
microvesicles have not
been described before
in BALF to the best of
our knowledge.
•Microvesicles appear
to lack the ability to
produce superoxide for
immune defense.
Time
Figure 1 key points:
Figure 2 key points:
Figure 3 key points:
• Pulmonary
microvesicles are
typically 2 microns in
size.
• They are roughly 20X
larger than previously
described pulmonary
exosomes, suggesting
they are an entirely
different particle.
• Xanthine and Xanthine
Oxidase are positive
controls for superoxide
production.
• Microvesicles at both
the 2 and 7 hour time
points do not show the
ability to produce
superoxide.
• A: Protein Standards
• B: 7 hour time point
• C: 4 hour time point
• D: 2 hour time point
• E: 0 hour time point
• F: Naïve
• Upregulation of a 67kD
protein is seen as time
increases in mice
inoculated with
Aspergillus fumigatus
•Proteins associated
with microvesicles are
upregulated as time
increases when the host
has been inoculated
with Aspergillus
fumigatus.
•Future analysis will
involve studying
recruitment of
microvesicles and
further characterization.