Response of Airway Cells to 20,000 EU of Endotoxin in Normal Adults
Response of Airway Cells to 20,000 EU of Endotoxin in Normal Adults
The purpose of this study is to determine the range of neutrophil response (influx of PMNs to
 the airways as determined in sputum) after inhalation of CCRE (20,000 EU).
Twenty-four or forty-eight hours (study day 1) prior to the inhalation challenge, subjects
 will undergo a physical examination of the ears, nose, throat and chest and will have an
 assessment of vital signs (temperature, pulse, respiratory rate, blood pressure), oxygen
 saturation, symptom score assessment and undergo spirometry to rule out acute illness prior
 to challenge. All female volunteers will undergo a urine pregnancy test. A similar
 examination will take place immediately prior to inhalation challenge with 20,000 EU of CCRE,
 as well as 30, 60, 120, 180, 240, 300 and 360 minutes and 24 hours after challenge (with the
 exception of a pregnancy test which will only be administered prior to sputum induction on
 study day 1). Induced sputum will be 24 to 48 hours prior to challenge, and again six hours
 after challenge on Study day 2. Sputum will be analyzed for PMN content, CD14 expression on
 airway macrophages and monocytes, soluble CD14 levels, cytokine levels in sputum and products
 of inflammatory cells (eosinophil, cationic protein, myeloperoxidase). Blood will be
 collected for a CBC and differential when the pre- challenge induced sputum is obtained and
 again 6 hours after challenge and 24 hours after challenge. Blood lymphocytes from a 10 ml
 aliquot will be assessed for lymphocyte subset and activation markers, as well as
 proliferative and cytokine responses. Blood will also be collected and will be assessed by
 whole blood assays for markers of inflammatory and immune activation (CD11b, CD14, CD64,
 CD16, HLA-DR, CD45, CD3, CD80, CD86, CD83, CD40) and function (phagocytosis and oxidative
 burst) by three color flow cytometry. Plasma will be separated from 4 ml and frozen at 70° C
 for future analysis of mediators of interest, such as cytokines, as well as other systemic
 effects of endotoxin.

 Subjects will then be discharged to home with contact information for study staff. If a
 volunteer has an unexpected complication such as shortness of breath, decrease in pulmonary
 function, unstable vital signs or other he or she will be admitted to the GCRC for overnight
 observation. The next morning (study day 3) subjects will return to the CEMALB for a final
 set of vital signs, spirometry, symptom scoring, a venipuncture and an examination by a study
 physician.

 Each volunteer will be given a symptom scoring sheet for each day up to 96 hours (4 days)
 after challenge.

 Each sheet will include the name and phone number of the study coordinator and study MD.
 Sample home symptom scoring sheets and instructions are included with this protocol.

 Between 48 and 96 hours after challenge, each volunteer will be called to determine their
 status, inquiring about symptoms, symptom scoring, and need for medication and/or physician
 visits.

 Between 7 and 10 days of the challenge dose, each subject will be asked to return for a study
 discontinuation visit. At that time temperature, pulse, systolic and diastolic BP,
 respiratory rate, FVC and FEV1 and SpO2 (oxygen saturation), and symptoms scores will be
 assessed and, if abnormal, medical evaluation as directed by the study physician will be
 undertaken.
Inclusion Criteria:

 -  Inclusion: Healthy, non-allergic, non-asthmatic 18-50 year old males and females with
 a < 0.5 pack year history of tobacco use who have a negative allergy skin test, normal
 lung function, defined as (Knudsen 1976/1984 predicted set):

 1. FVC of > 80 % of that predicted for gender, ethnicity, age and height

 2. FEV1 of > 80 % of that predicted for gender, ethnicity, age and height

 3. FEV1/FVC ratio of > 75% of that predicted for gender, ethnicity , age and height

 -  Oxygen saturation of > 94 %,

 -  Normal blood pressure (Systolic between 150 - 90, Diastolic between 90-60 mm Hg),

 -  Symptom Score no greater than 6 (out of a possible 24) for total symptom score with a
 value no greater than 2 for any one score. No score may be greater than 2,

 -  Negative methacholine inhalation challenge.

 -  On the day of a challenge, body temperature must be no greater than 37.8 degrees,
 measured orally.

 Exclusion Criteria:

 -  A history of significant chronic illnesses (to include diabetes, autoimmune diseases,
 immunodeficiency state, known ischemic heart disease, chronic respiratory diseases
 such as chronic obstructive pulmonary disease or asthma, hypertension).

 -  Allergy to any medications which may be used in the course of this study (albuterol,
 acetaminophen, aspirin or non-steroidal anti-inflammatory agents, corticosteroids,
 lactose).

 -  Positive pregnancy test within 48 hours of the time of challenge.

 -  Medications which may impact the results of the endotoxin challenge, interfere with
 any other medications potentially used in the study (to include steroids, beta
 antagonists, non-steroidal anti-inflammatory agents) or suggest an ongoing illness
 (such as antibiotics).

 -  Acute, non-chronic, medical conditions, including (but not limited to) pneumonia or
 bronchitis requiring antibiotics, febrile illnesses, flu-like symptoms must be totally
 resolved symptomatically for 2 weeks.

 -  Unspecified illnesses, which in the judgment of the investigator increase the risk
 associated with endotoxin inhalation challenge, will be a basis for exclusion.

 -  Persons employed within the past 6 months in an occupation with high risk for
 endotoxin exposure (specifically persons working in a swine confinement facility,
 cotton storage or grain storage site)