In Vitro Basophil Responsiveness to Allergen Challenge After Gamma-tocopherol Supplementation in Allergic Asthmatics
In Vitro Basophil Responsiveness to Allergen Challenge After Gamma-tocopherol Supplementation in Allergic Asthmatics
Purpose: This is a non-masked study with a primary endpoint of in vitro basophil activation
 by the allergen D. farinae, comparing basophil activation before and after seven days of
 supplementation. Secondary endpoints will include circulating antioxidant levels (tocopherols
 and metabolites), in vitro basophil activation to IgG anti-IgE and to
 N-formyl-methionyl-leucyl-phenylalanine (f-met-leu-phe), and monocyte and basophil
 responsiveness to in vitro endotoxin challenge.

 Participants: Twenty allergic asthmatic volunteers Procedures (methods): Volunteers will be
 given 1200 mg of a gamma tocopherol enriched supplement, a commercially available supplement
 form of vitamin E. Study participants will undergo assessment of general health, lung
 function assessment, symptom scoring, and epicutaneous skin test to allergens at baseline and
 after supplementation. Blood samples will be collected at baseline and after 7 days of
 gamma-T treatment.
Oxidant stress plays an important role in mucosal inflammation such as seen with asthma.
 Previous studies have also shown that asthmatic individuals tend to have lower endogenous
 levels of antioxidant such as vitamins E and C, and that supplementing antioxidants can
 decrease exacerbations associated with ozone exposure in children. We are interested in
 future studies to examine the benefits of gamma-tocopherol supplementation for people with
 allergic asthma. This current study will determine if gamma tocopherol supplementation
 reduces allergic responses in vitro, providing us information that will allow us to design
 future masked placebo-controlled studies of this potentially important antioxidant on in vivo
 allergic responses.

 The purpose of this study is to address the question if in vivo gamma-tocopherol
 supplementation at 1200 mg daily blunts allergen-specific inflammatory responses in vitro.
 Members of our group have previously shown that administration of 100 mg/kg of gamma
 tocopherol daily for four days prior to Ova challenge in sensitized allergic brown Norway
 rats prevented eosinophil infiltration into the airways1. In addition, this dose of gamma
 tocopherol blunted production of IL-4, IL-5, IL-13, IFN-gamma in the nasal airway; and PGE2,
 LTB4 and cysteinyl leukotrienes by the pulmonary airway. Mucous cell metaplasia was decreased
 as well in the gamma Tocopherol treatment group 1. In an in vivo study of gamma tocopherol
 performed at our center (IRB# 05-CEMALB-1407), we found that daily administration of 2
 capsules of a gamma tocopherol rich preparation (each capsule containing 623 mg of gamma
 tocopherol, 61.1 mg of d-alpha-tocopherol, and 11.1 mg of d-beta tocopherol), was able to
 increase serum levels of gamma tocopherol to 18.6 + 2.6 uM after 8 days of daily
 administration; serum levels of alpha tocopherol were 25.2 + 2.4 uM, and delta-tocopherol
 were 5.1 + 1.1 uM2 . Using the data from the in vivo study, we performed basophil activation
 tests on dust mite allergic subjects, pretreating blood obtained from venipuncture with
 pharmacologic doses attained in the vivo study with gT, aT, gCEHC, and aCEHC. We found that
 gT, gCEHC, and aCEHC blunted basophil activation induced by the allergen D. farinae, as
 measured by upregulation of CD63 on the cell surface of basophils. As secondary aims, we will
 also examine the effect of gamma tocopherol supplementation on non-allergic stimuli that have
 been shown to activate basophils, such as IgG anti-IgE and
 N-formyl-methionyl-leucyl-phenylalanine (f-met-leu-phe), as well as in vivo allergen-specific
 responsiveness through epicutaneous skin testing before and after supplementation.
 Exploratory analyses will include assessing the effect of gamma tocopherol supplementation on
 monocyte and basophil responsiveness to in vitro endotoxin and lipoteichoic acid challenge.
Inclusion Criteria:

 -  1. Specific allergy to House Dust Mite farinae confirmed by positive immediate skin
 test response.

 2. Oxygen saturation of > 94 % at baseline 3. Blood pressure within the following
 parameters (Systolic between 140 - 90, Diastolic between 90-60 mm Hg) 4. Moderate or
 severe persistent asthma according to NHLBI definitions including history of one of
 the following:

 1. Episodic wheezing, chest tightness or shortness of breath consistent with asthma
 occurring at least once a week that may affect activity

 2. Asthma symptoms occurring at night or during sleep at least 1 time per week

 3. measured FEV1 or FVC is <80% of predicted OR

 4. Physician diagnosed moderate or severe persistent asthma which is currently
 treated or controlled with maintenance medication including moderate or high dose
 inhaled corticosteroid, or any dose of inhaled corticosteroid and a long-acting
 inhaled B2-agonist

 Exclusion Criteria:

 -  1. Any chronic medical condition considered by the PI as a contraindication to
 receiving gamma-T, including significant cardiovascular disease, diabetes requiring
 medication, chronic renal disease, chronic thyroid disease, kidney disease or
 coagulation defects.

 2. Use of inhaled steroids, cromolyn or leukotriene inhibitors (Montelukast or
 Zafirlukast) which have been used for at least one month are allowed. Patients must be
 on a stable regimen of maintenance asthma therapy which has not changed in the past
 month prior to entrance into the study.

 3. Non-steroidal Anti-Inflammatory Drugs (NSAIDS) or aspirin (ASA) use within 48 hours
 of beginning the study, and inability to suspended use of these medications during the
 length of the study.

 4. Use of anticoagulants including warfarin, heparin, or clopidogrel. 5. Diagnosis of
 anemia or abnormal blood counts at screening. Abnormal PT or PTT values at screening
 (PT that is prolonged more than 1-2 sec and PTT prolonged more than 3-5 sec of normal.
 The normal values used will be those defined by McLendon lab.) 6. Pregnancy or nursing
 a baby. As this is a phase I study, the potential risk to a fetus cannot be justified.

 7. Children will not be included in this study as the potential risk to a growing
 child cannot be justified.

 8. Adults age 51 and older are excluded as the potential for concomitant illness in
 this population increases the risk for confounding the data.

 9. Known vagal response to venipuncture 10. Use of any tobacco product within the past
 6 months 11. Hypertension, classified as a systolic blood pressure of equal to or
 greater than 140, and a diastolic blood pressure equal to or greater than 90.