Biotechnology Applications
The 1950’s
GM Firebird III
Public Health - Sanitation
Public Health - Vaccination
Polio
Allergy and Asthma Epidemic in Developing
Countries
Devereux G. 2006. Nature Rev Immunol 6;869-874.
Hygiene Hypothesis
early childhood infections inhibit the tendency to develop allgergic disease
Helminth Infections & Allergies: Inverse Correlation
Helminth infections are associated with a decreased
incidence of allergic disease in industrialized areas
% Population* Helminth Allergic Disease***
Infection**
Globally ~ 25% ~ 2%
United States 5,500,000
features per chip
Image Courtesy of Bioteach
Innate Source of Th2 Cytokines in the Lung
lung intestine lung intestine
Nb Infection Innately Induced AAM Genes
lung intestine lung intestine
Polymerase Chain Reaction - PCR
Thermal
Vents
Thermus aquaticus
T. aquaticus
DNA Polymerase
30 rounds of amplification = >1,000,000,000 molecules
Real Time RT PCR results
Alternatively Activated Macrophage
Alternatively
Activated
Macrophages
T Cell Help is Required for AAAM Maintenance
Observations:
Nb induces cellular/molecular changes through
day 12 PI
Helminth-induced suppression of reactivity to
secondary challenge (Wohlleben et al. 2004; Wilson
et al. 2005; Yang et al. 2006)
Hypotheses:
• Helminth infection results in permanent
alterations to the lung’s immunological
environment
Nb Induces an Altered Lung Environment
Experimental Design
2-4 13-15 36
Granular Macrophages Contain Surfactant
day 36 PI
Flow cytometry
antibody-based
detection of
surface molecules
Day 36 PI AMs are CD11c+, F4/80+, Mrc1mid & MHC IIhi
day 36 PI
Surface Phenotype of CD11c+ AAAM
Day 0 2 4 8 13
MHC I
CD11c+ AAAM up-regulate: MHC II
MHC I & II PDL-I
PDL-1 PDL-2
CD80/CD86 CD80
CD40 CD86
CD1d
CD40
ICAM-1
CD1d
AAAM are Highly Phagocytic
Nb+ 0.02 Nb- 0.07
Ova-
CD11c APC CD11c APC
22.5 9.2
Ova+*
CD11c APC CD11c APC
5.2 3.5
Ova+*
CytoB Ova-Alexa 448 at 2
μg/ml
for 30 min
CD11c APC CD11c APC CytoB - cytochalasin B
at 800 μg/ml
CD11c+ AM Expression Profile
AAM (arg1, fizz1,
Ym1/2, trem2b)
Up-regulated >2-fold
Scavenger
Receptors
(clecsf5/6/8/10, marco, 194 797
mgl1/2, msr1/2) day 2 PI 76 day 4 PI
Chemokines
(606) (1,674)
(ccl4/8/9/11/12/17/24)
277
59 524
Cell Cycle
Lipid Metabolism
(ppar gamma, cd68,
apobec1, ch25h)
392
day 8 PI
(1,252)
Altered Lung Environment:
Hypotheses:
• The Nb-altered lung environment
results in a modulation in responses
to secondary challenges - HDM
allergen challenge
• Gene array, PCR, histology,
lung function
Similar Numbers, Different Genes are Up-regulated
Nb-Altered Lung Down-Regulates Many More Genes
Nb-Altered Lung Decreases Airway Hyperresponsiveness
Altered Lung Environment:
Dampened Responses to Allergen
Key Findings:
• Persistent changes resulted in a reduction in
the overall inflammatory response to allergen
challenge in the lungs of Nb-infected animals
• Distinct gene expression profiles in the lungs
of NbHDM vs HDM mice
• Lower fold-increase of transcription of Th1 &
Th2 cytokines in NbHDM mice
• Resistance in the airways is significantly
depressed in the lungs of NbHDM mice
Hygiene Hypothesis
early childhood infections inhibit the tendency to develop allgergic disease
Alternatively Activated Alveolar Macrophages
(PDL 1/2 ?)
CD1d
?
NKT IL-6
IL-21
?
Acknowledgments
Scott Lab
Josh Reece JHU Gene Array Core Facility
Mark Siracusa Anne Jedlicka
Kiwon Park Meg Mintz
Sarah Coffman Jason Bailey
Ellen Tarr Amanda Dziedzic
JHU
Sabra Klein
Mike Overstreet Collaborators
Fidel Zavala Joe Urban (USDA)
Mark Soloski Andrea Keane-Myers (NIH)
Bill Spannhake Jamie Lee (Mayo Clinic)
Wayne Mitzner
Jon Fallica
Doug Norris
Vern Carruthers
Funding / Support
Brian Schofield NIH NHLBI (U01 HL66623)
Paul Fallon NIH Training Grant (T32AI007417)
Theresa Southard Hegner, Cort Root Memorial Scholarship
Martin Frobisher Fellowship
Cory Brayton