Systemic Inflammatory Response to CCRE

This is a single center, screening study involving recruitment of 18 normal volunteers (NV).
The protocol is powered to compare absolute neutrophil count (ANC) in the blood 6 hours after
inhalational challenge with CCRE compared baseline. In addition, as secondary endpoints, the
investigators will evaluate inflammatory cytokine levels in systemic circulation and access
for the presence of genes thought to be related to endotoxin response. Visits will be
conducted by a study coordinator or other study staff, physical exams will be performed by
the PI or other study physician.

Eighteen subjects will be recruited. There will be no gender or ethnic restrictions, and
subjects will be healthy volunteers. Prior to enrollment in this study, subjects will have
participated in our protocol 98-0799 (screening and database study for the CEMALB). Data such
as medical history and allergy skin testing collected during the screening protocol will be
included with data in this study. Subjects with positive allergy skin testing will not be
excluded. Spirometry will be performed to determine the current level of lung function.

In order to identify those individuals who appear more susceptible to the systemic
inflammatory effects of endotoxin exposure, subjects will undergo inhaled endotoxin challenge
at the baseline visit followed 6 hours later by blood draw for complete blood count with
differential to allow for calculation of absolute neutrophil count (ANC). Subjects will be
considered "responders" if their peripheral ANC increases by at least 20% following inhaled
endotoxin challenge.

Inclusion Criteria:

1. Subjects must be 18-50 years of age to be eligible for study participation

2. Subjects must be willing to and able to provide informed consent and participate in
all study procedures

3. Normal lung function, defined as (Knudson 1976/1984 predicted set):

- 1. Forced Vital Capacity (FVC) of > 80% of that predicted for gender, ethnicity,
age and height

- 2. Forced Expiratory Volume in the first second (FEV1) of > 80% of that predicted
for gender, ethnicity, age and height

- 3. FEV1/FVC ratio of > 0.75 of that predicted for gender, ethnicity, age and
height

4. Oxygen saturation of > 94% Normal blood pressure (Systolic between 150 - 90, Diastolic
between 90-60 mm Hg)

5. Symptom Score (defined in section "f") no greater than 6 (out of a possible 24) for
total symptom score with a value no greater than 2 for any one score.

Exclusion Criteria:

1. Any chronic medical condition considered by the PI as a contraindication to the
exposure study including significant cardiovascular disease, diabetes requiring
medication, chronic renal disease, or chronic thyroid disease.

2. Physician directed emergency treatment for asthma exacerbation within the preceding 3
months.

3. Moderate or Severe asthma

4. Exacerbation of asthma more than 2x/week that would be characteristic of a person of
moderate or severe persistent asthma as outlined in the current NHLBI guidelines for
diagnosis and management of asthma.

5. Nighttime symptoms of cough or wheeze greater than 1x/week at baseline (not during a
clearly recognized viral induced asthma exacerbation) which would be characteristic of
a person of moderate or severe persistent asthma as outlined in the current NHLBI
guidelines for diagnosis and management of asthma

6. Daily requirement for albuterol due to asthma symptoms (cough, wheeze, chest
tightness) which would be characteristic of a person of moderate or severe persistent
asthma as outlined in the current NHLBI guidelines for diagnosis and management of
asthma. (Not to include prophylactic use of albuterol prior to exercise).

7. History of intubation for asthma

8. Daily use of NSAIDs, or inability to withhold NSAIDs for 4 days prior to dosing.

9. Use of medications that may impact the results of the study to include, but not
limited to, systemic corticosteroids, beta blockers.

10. Cigarette smoking > 1 pack per month.

11. BMI>35.

12. Pregnant or breast feeding women will not be included.

13. Subjects who are employed within the past 6 months in an occupation with high risk for
endotoxin exposure, such as grain storage sites or swine containment.

14. Subjects will be deferred after any acute, non-chronic medical condition requiring
treatment, such as bronchitis, pneumonia or febrile illness for a minimum of 4 weeks
after complete resolution of symptoms.