Exposure to agriculture dust particles during allergen sensitization exacerbates allergic
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Exposure to agriculture dust particles during allergen sensitization exacerbates allergic airway inflammation. Alex Castañeda, M.S. Graduate Group in Immunology University of California Davis Dr. Kent Pinkerton Director, Center for Health & the Environment

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Alex Castañeda, M.S. Graduate Group in Immunology University of California Davis

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Alex casta eda m s graduate group in immunology university of california davis

Exposure to agriculture dust particles during allergen sensitization exacerbates allergic airway inflammation

Alex Castañeda, M.S.

Graduate Group in Immunology

University of California Davis

Dr. Kent Pinkerton

Director, Center for Health & the Environment

Department of Pediatrics, School of Medicine

Professor of Anatomy, Physiology and Cell Biology

UC Davis School of Veterinary Medicine


Asthma

Asthma

Environmental/Occupational

Factors

Allergen

Ovalbumin

Ambient Particles

Genetic Factors

Kim et al. 2010. The many pathways to asthma. Nature Immunology 11: 577-584


Allergic asthma

Allergic Asthma

Symptoms: Airway inflammation, bronchial constriction, coughing, wheezing, and shortness of breath

Prevalence: 235 million people worldwide (11 million in the US)

In California (San Joaquin Valley) 25% of all children are diagnosed with allergic asthma


Particle pollution

Particle Pollution

Studies have shown a correlation between asthma and high particulate matter exposure

-Particular matter exacerbate asthma

-The use of source specific ambient particles has not been studied

Particulate Matter:

Vehicle emissions (gasoline)

Agricultural Dusts

Diesel exhaust gases

Mineral dusts

Hydrocarbons

Particles Size:

Coarse particles

Fine Particles

Ultrafine particles (0.1µm in diameter)

Location:

Fresno California (Leading agriculture-producing county in the nation)

Particles collected during summer nights (2010)


Agricultural dust

Agricultural Dust

Generation

Field preparation for crops, fertilization, crop harvest (ex- almonds)

Composition

Inorganic (Mineral Dusts)

Silcates (aluminum silicates), quartz, transition metals

Organic (Carbonates)

Toxicity of Inorganic Agricultural Dusts

Cellular toxicity

Generate free radicals

Oxidative cellular stress

Activation of the immune system towards an inflammatory state

-Fibrogenesis


Hypothesis

Hypothesis

Exposure to agricultural dust particles during sensitization with an allergen (house dust mites or ovalbumin) will exacerbate airway inflammation in BALB/c mice


Methods

Methods

Groups

1 -Control (No Allergen + Filtered Air)

2- OVA + Filtered Air

3- OVA + Particulate Matter

4- pOVA + Filtered Air

5- pOVA + Particulate Matter

6- pHDM + Filtered Air

7- pHDM + Particulate Matter

Tissue Collection

Blood Plasma (IgE,IgG1)

BALF

Lung (Homogenize & Fixed)

Murine Model- BALB/c mice

Treatments

Allergen- OVA, Purified OVA or

Purified House Dust Mites (HDM)

Agricultural Dust- Ultrafine Particles (Fresno)

Sensitization (days 1, 3, & 5)

OVA & pOVA- 10ug intranasal

HDM- 25ug intranasal

Challenge (days 12, 13, & 14)

OVA & pOVA- 10ug aerosolized

HDM- 10 ug OVA

Allergen Challenge

Allergen Sensitization+

Ambient Particles

0 5 10 15

Necropsy


Balf cellular proflies

BALF Cellular Proflies

Macrophages

Total Cells

Eosinophils

Neutrophils


Results tissue histology

Results- Tissue Histology

Control Allergen and particulate matter

Alcian Blue Periodic Acid Schiff’s Stain

Mucosubstance


Results tissue histology1

Results- Tissue Histology

P

M

D

Volume of Intraepithelial Mucosubstances

(

µm3

µm2

)

Airway Inflammation:

Area of Basal Lamina


Results tissue histology2

Results- Tissue Histology

P

M

D


Results total protein

Results- Total Protein


Results antibody production

Results- Antibody Production


Alex casta eda m s graduate group in immunology university of california davis

Results- IL-17-A Production

Barnes et al. 2008. Immunology of asthma and COPD.Nature Reviews Immunology 8, 183-192


Alex casta eda m s graduate group in immunology university of california davis

Results- IL-17-E Production


Summary

Summary

Airway inflammation was exacerbated when particulate matter was introduced during allergen sensitization (HDM only, not OVA)

-Significantly elevated levels of total cells, macrophages, and neutrophils

Based onmuco-substance production HDMA causes significantly more airway inflammation over Ova

IL-17A production was elevated in the HDMA model, correlating with increased neutrophils, however there is no correlation to particles


Alex casta eda m s graduate group in immunology university of california davis

Future Work

Investigate the mechanisms through which agricultural dust enhances airway inflammation

-How particles activate the immune system?

Investigate neutrophil infiltration-Do particles increase neutrophilic inflammation?


Acknowledgements

Acknowledgements

Pinkerton Laboratory UC Davis Western Center for Agricultural Health and Safety

Kent Pinkerton, Ph.D.Marc Schenker, M.D.

Chris CarosinoSandra Freeland

Dale UyeminamiAll other members

Janice Peake

Katherine JohnsonGraduate Group in Immunology

Imelda Espiritu

Alexa Pham

Esther Shin

Jocelyn Claude

All other members


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