HIV+ Alveolar Macrophage Oxidant-mediated Apoptosis of Pulmonary Endothelium

The purpose of this study is to analyze biologic samples from the blood, airways and/or urine
of HIV- and HIV+ smokers and non-smokers to better understand the etiology and pathogenesis
of emphysema in association with HIV infection. The underlying hypothesis is that the
combination of HIV infection and smoking creates chronic oxidant stress in the lower
respiratory tract that promotes cellular loss and contributes to the progressive development
of emphysema. Preliminary data strongly supports our hypothesis that the accelerated
development of emphysema among HIV+ smokers is due in part to the interaction of HIV directly
on the macrophage found in the pulmonary alveolus. The interaction causes a release of
exaggerated amounts of oxidants in the lower respiratory tract and leads to increased levels
of oxidized metabolites. HIV+ individuals with emphysema have high plasma levels of apoptotic
pulmonary capillary-derived micro particles that contain oxidized metabolites. The increased
release of micro particles is characteristic of an apoptotic process.

We will study this hypothesis by sampling HIV+ and HIV‾ subjects alveolar macrophages (AM),
which are found on the epithelial surface of the lung, and epithelial lining fluid (ELF)
found in the lower respiratory tract. We will also assess plasma pulmonary capillary-derived
endothelial microparticles (EMPs) as a biomarker for pulmonary apoptosis. Using newly
developed mass spectrometry methodologies, we will quantify the oxidant stress of AM, ELF and
plasma EMPs, and identify specific oxidized metabolites within each of these compartments.
Finally, we will examine the interaction in vitro to tease apart the contribution of each
component (AM, ELF, and plasma EMPs) of the interaction.

To assess this concept, the following aims will be addressed:

Specific Aim 1 (n=160). To explore the extent of the oxidant stress in the lower respiratory
tract in association with HIV infection and smoking.

Specific Aim 2 (n=160). To evaluate plasma levels of capillary apoptosis and oxidation state
of HIV+ nonsmokers and smokers.

Specific Aim 3 (n=160). To examine the interaction of pulmonary capillary endothelium and
various oxidant effector components to identify oxidant-vulnerable pathways relevant to the
development of new treatment therapies.

Inclusion Criteria:

HEALTHY VOLUNTEER RESEARCH SUBJECTS

- All study subjects should be able to provide informed consent

- Males or females ages 18 years and older

- Must provide HIV informed consent

VOLUNTEER RESEARCH SUBJECTS WITH LUNG DISEASE

- Must provide informed consent

- Males and females age 18 years and older

- Lung disease proven by at least one of the following: symptoms consistent with
pulmonary disease; (2) chest X-rays consistent with lung disease; (3) pulmonary
function tests consistent with lung disease; (4) lung biopsy consistent with lung
disease; (5) family history of lung disease; and/or (6) diseases of organs with known
association with lung disease

- Must provide HIV informed consent

Exclusion Criteria:

HEALTHY VOLUNTEER RESEARCH SUBJECTS

- Individuals not deemed in good overall health by the investigator will not be accepted
into the study.

- Habitual use of drugs and/or alcohol within the past six months (Acceptable: -
Marijuana one time in three months; average of two alcoholic beverages per day; drug
and/or alcohol abuse is defined as per the DSM-IV Substance Abuse Criteria).

- Individuals with history of chronic lung disease, including asthma or with recurrent
or recent (within three months) acute pulmonary disease will not be accepted into the
study.

- Individuals with allergies to atropine or any local anesthetic will not be accepted
into the study.

- Individuals with allergies to pilocarpine, isoproterenol, terbutaline, atropine or
aminophylline will not be accepted into the study.

- Females who are pregnant or nursing will not be accepted into the study

VOLUNTEER RESEARCH SUBJECTS WITH LUNG DISEASE

- Any history of allergies to xylocaine, lidocaine, versed, valium, atropine,
pilocarpine, isoproterenol, terbutaline, aminophylline, or any local anesthetic will
not be included in the study.

- Habitual use of drugs and/or alcohol within the past six months (Acceptable: Marijuana
one time in three months; average of two alcoholic beverages per day; drug and/or
alcohol abuse is defined as per the DSM-IV Substance Abuse Criteria)

- Females who are pregnant or nursing