Effects of Electronic Cigarettes on Lung Biology

The specific aim of this study is to assess the hypothesis that electronic smoking disorders
the biology of the airway epithelium and alveolar macrophages, to evaluate these changes in
the lungs of normal nonsmokers exposed to electronic cigarettes, and to directly determine if
the inhaled smoke from electronic cigarettes disturbs the biology of the naïve lung that has
not been exposed to tobacco products. Because nicotine can be addictive with continual use,
we will limit the exposure to a maximum of a nicotine equivalent of 1/2 pack of cigarettes
over a 5 day period, a level that has been used safely in studies of healthy nonsmokers'
naïve to nicotine.

To obtain the most unbiased data about the effect of electronic cigarette use on the lung,
normal nonsmokers will be briefly exposed to electronic cigarettes and lung cells, airway
epithelium and alveolar macrophages, lung epithelial lining fluid (ELF) and serum will be
collected before and after e-cigarette exposure. There will be 5 study groups consisting of
10 non-smoker subjects (A-E), with each group undergoing an increased exposure to the amount
of e-cigarettes (Group a being the lowest exposure, group E being the highest). Six of the
subjects in each group will be exposed to e-cigarettes with nicotine while the remaining four
in each group being exposed to nicotine-free e-cigarettes. Subjects will be assigned to use
electronic cigarettes that either do or do not contain nicotine in a randomized, blinded
manner.

The duration of which the subjects will be observed for e-cigarette exposure will be 8 days.
Baselines for each group will be established on Day 1 in which all subjects will undergo a
bronchoscopy and an assessment of vital signs.

For subjects in group A, at the day 8 visit after the vital signs are assessed, 3 hours prior
to bronchoscopy, the subject will be asked to inhale 10 puffs of a "Blu" brand e-cigarette
(for the nicotine-containing e-cigarette, this is approximately equivalent to < 1 cigarette).
At 2 hours prior to the bronchoscopy procedure, if the subject has no adverse effect to the
e-cigarette he/she will again be asked to inhale 10 puffs of the "Blu" e-cigarette. The vital
signs will again be confirmed and any symptoms assessed prior to proceeding with
bronchoscopy. Subjects in group B will undergo an identical exposure, but on both days 7 and
8. Subjects in group C will undergo similar exposure on days 6-8, group D on days 5-8, and
group E days 4-8 for a maximum of 5 exposure days prior to bronchoscopy. The research team
will contact subjects at 1 month and 6 months after their Day 8 bronchoscopy in order to
determine whether or not they have become dependent on nicotine.

Inclusion Criteria:

- Males and females, age 21 or older

- Must be capable of providing informed consent

- Self-reported never-smokers, with current smoking status validated by the absence of
nicotine metabolites in urine (nicotine <2 ng/ml and cotinine <5 ng/ml)

- Normal physical examination

- Good overall health without history of chronic lung disease, including asthma, and
without recurrent or recent (within 3 months) acute pulmonary disease

- Normal routine laboratory evaluation, including general hematologic studies, general
serologic/immunologic studies, general biochemical analyses, and urine analysis

- Not taking any medications relevant to lung disease or having an effect on the airway
epithelium

- Negative HIV serology

- Normal chest X-ray (PA and lateral)

- Normal electrocardiogram (sinus bradycardia, premature atrial contractions are
permissible)

- Females - not pregnant

- No history of allergies to medications to be used in the bronchoscopy procedure

- Willingness to participate in the study

Exclusion Criteria:

- Unable to meet the inclusion criteria

- Current active infection or acute illness of any kind

- Evidence of malignancy within the past 5 years

- Current alcohol or drug abuse

- Pregnancy