AB198 Abstracts
FEBRUARY 2010
774 In Vitro Immunomodulatory Effect Of ASHMI (Anti-asthma
Herbal Medicine Intervention) On PBMCs From Asthmatics
S. P. Patil, W. J. Ji, A. Singha, D. E. Hendricks, P. J. Busse, H. A. Samp-
son, J. P. Wisnivesky, X. Li; Mount Sinai School of Medicine, New York,
NY.
RATIONALE: ASHMI has been shown to be safe and have anti-asthma
effects in clinical studies. However, its immunological effects on peripheral
blood mononuclear cells (PBMCs) of patients has not been evaluated. The
goal of this study was to investigate the immunological effects of ASHMI
on asthma patient’s PBMCs enrolled in an on going phase II clinical trial.
METHODS: PBMCs (4 x 105) from patients (Allergen, n515; PHA,
n510) enrolled in an ASHMI Phase-II clinical trial were obtained at the
baseline visit and cultured in AIM-V media alone, with allergen (200mg/
mL), allergen+ASHMI (250mg/mL), PHA (2mg/mL) or PHA+ASHMI
(250mg/mL). The allergens used were house dust mite, cat pelt, dog dander,
tree mix, mold mix, ragweed and cockroach. After 7 days supernatants
were analyzed for IL-5, IFN-g and IL-10 levels by ELISA. In additional
experiments, anti- IL-10 neutralizing antibody (50ng/mL) or isotype con-
trol Ab. were added to the cultures.
RESULTS: ASHMI at 250mg/mL was optimal (cell viability > 90%). IL-5
levels in allergen+ASHMI [125.03pg/mL(6-1000.08)] and PHA+ASHMI
[263.35pg/mL(98.94-347.99)] supernatants were significantly reduced
(p< 0.05) as compared to allergen [504.42pg/mL(18-1086.26)] or PHA
[578.30pg/mL(243.27-760.84)] only. IFN-g production was not affected
in allergen+ASHMI [347.96pg/mL(12-5239.61)] or PHA+ASHMI
[8770.86pg/mL(2951.94-21845.62)] cultures as compared to allergen
[765.89pg/mL(12.49-6301.76)] or PHA [8390.82pg/mL(2991.59-
16949.42)] alone. Production of IL-10 was increased in
allergen+ASHMI [23.1pg/mL(6.66-74.11)] or PHA+ASHMI [379.95pg/
mL(173.25-1274.64)] supernatants (p< 0.05) as compared to allergen
[11.54pg/mL(4.7-26.71)] or PHA [185.5pg/mL(113.69-486.51)] only.
Anti-IL-10 did not affect ASHMI mediated suppression of IL-5 levels.
CONCLUSIONS: ASHMI reduces IL-5 production by PBMCs from
asthma patient’s in vitro, which may not be associated with IL-10 production.
775 Perception of Traditional Medication and Alternative
Therapies for Asthma in Relationship to Race and Ethnicity
in Inner-City School Children
A. Landrum1, J. M. Gaffin1,2, J. Oviedo3, A. Bailey3, W. Phipatanakul1,2;
1
Harvard Medical School, Boston, MA, 2Childrens Hospital Boston, Bos-
ton, MA, 3Channing Laboratory, Boston, MA.
RATIONALE: Many patients with asthma try alternative treatments in ad-
dition to or in lieu of traditional therapies. Our goal was to determine if dis-
parate perceptions of traditional and alternative asthma therapies exist
between racial/ethnic groups.
METHODS: 83 elementary inner-city students with asthma were sur-
veyed by parental questionnaire. Questions were asked regarding tradi-
tional medication use, perception of effectiveness of traditional
therapies, and use of alternative therapies. Prevalence ratios were calcu-
lated to examine the effect of race/ethnicity on the outcomes of interest.
Fishers exact test determined significance.
RESULTS: There were 46 (55%) Hispanic, 20 (24%) Black and 17 (21%)
Mixed/Other students surveyed. All of the Black students believed tradi-
tional medications controlled their asthma, while only 70% of Hispanic
TUESDAY
students did. The prevalence ratio comparing BlacksÕ and HispanicsÕ per-
ception of asthma control by traditional medications significantly favored
Blacks by a factor of 1.4 (95% confidence interval 1.2 - 1.7; P value <0.01).
None of the parents reported stopping traditional asthma medications. 20%
of Hispanics reported using alternative remedies such as teas, rubs, and
herbs for asthma, compared to 5% of Black students. 4% of Hispanics re-
ported using alternative therapies instead of traditional medications, while
none of the Black students did.
CONCLUSIONS: A significantly smaller proportion of Hispanic students
than Black students perceived traditional asthma medication to control
their asthma. More Hispanics reported using alternative therapies in addi-
tion to and in lieu of traditional therapies, though these did not reach
J ALLERGY CLIN IMMUNOL
statistical significance. Further studies are needed to fully understand
racial/ethnic differences in attitudes towards asthma therapy.
776 Flavonoids from Gan-Cao (Radix Glycerrhizae) Inhibit Th2
Memory Cell Cytokine Production
N. Yang, J. Zhu-ge, W. Zhang, B. Jayaprakasam, J. Goldfarb, H. Sampson,
X. Li; MountSinai School of Medicine, New York City, NY.
RATIONALE: Flavonoids, naturally occurring compounds in fruit, vegeta-
ble and other plants, are known to have anti-inflammatory properties. Several
flavonoids were isolated from Gan-Cao (Radix Glycerrhizae), one of the
herbs in the ASHMI formula. Previous data showed that ASHMI modulated
the Th2 response in asthma patients and a murine asthma model. In this study,
we investigated the effect of flavonoids isolated from Gan-Cao on memory
Th2 cell IL-4 secretion and their underlying mechanisms of action.
METHODS: Each flavonoid was characterized by a Liquid
Chromatography Mass Spectrometry system. Conalbumin-stimulated D 10
cells (a classical Th2 clone) were cultured in the presence of irradiated
AKR mouse spleen cells, and treated with several concentrations of each fla-
vonoid. After 72 hr, the concentrations of IL-4 in the supernatants were mea-
sured by ELISA. Cell proliferation was measured by [3H]-Thymidine
incorporation. GATA-3 protein levels were determined by Western Blot.
RESULTS: The flavonoids isoliquiritigenin, liquiritigenin and 7,4Õ dihy-
droxyflavone significantly suppressed secretion of IL-4 in a dose dependent
manner and the IC50 values were 10.91mg/mL, 18.85mg/mL, and 1.80 mg/
mL respectively. No toxicity was detected at any doses using trypan blue ex-
clusion and MTT assays. Real-time PCR results showed that IL-4 mRNA
levels were significantly reduced by 7,4Õdihydroxyflavone. This compound
also suppressed synthesis of the Th2-specific transcription factor, GATA-3.
CONCLUSIONS: 7,4Õ dihydroxyflavone exhibited the greatest suppres-
sion of the Th2 cytokines generated by effector memory Th2 cells. This
study demonstrated that Gan-Cao flavonoids may provide a potential
therapeutic approach for asthma and allergy.
777 A Novel Submicron Formulation of Nebulized Budesonide
Delivered by a Vibrating Mesh Nebulizer Significantly
Improves Delivery Efficiency at One-Fifth of the Nominal
Dose of both Pulmicort RespulesÒ and a Generic Budesonide
Suspensions Delivered via a Jet Nebulizer - A Potentially
Important Advancement in Pediatric Asthma
A. Bosco, M. Castillo, M. Conde, M. Luangvala; MAP Pharmaceuticals
Inc, Mountain View, CA.
RATIONALE: Innovative aerosol generators can improve nebulization
efficiency of aqueous suspensions by eliminating large residual volumes
and concentrating effects associated with jet nebulizers. A novel submi-
cron budesonide dispersion (sBUD) has been developed for use with these
innovative aerosol generators, which are not approved for use with mar-
keted aqueous suspensions used to treat of pediatric asthma.
METHODS: Using the AeroNeb GO (AN-GO) vibrating membrane nebu-
lizer (Aerogen Ltd., Ireland), the emitted mass (EM) of budesonide delivered
in 1 min from a low-volume dose of sBUD (MAP Pharmaceuticals, USA; 45
mcg/0.5mL, 90 mcg/0.5mL) was compared to Pulmicort RespulesÒ (PR;
AstraZeneca, USA) and a generic budesonide inhalation suspension (gBIS;
TEVA, USA) delivered by a Pari LC Plus jet nebulizer (250 mcg/2mL,
500mcg/2mL each) using a 60s pediatric tidal breathing pattern.
RESULTS: At one-fifth the nominal dose, 45 mcg of sBUD delivered in
0.5 mL with the AN-GO generated a significantly greater EM to the mouth
in 1 min compared to 250 mcg of PR and gBIS delivered in 2 mL by a jet
nebulizer (1262 vs. 762 vs. 661 mcg respectively). The same significant
difference was observed when 90 mcg of sBUD was compared to 500 mcg
of PR and gBIS (2162 vs 1362 vs 1162 mcg respectively).
CONCLUSIONS: sBUD delivered with the AN-GO provides a more por-
table and efficient delivery system for nebulized corticosteroid therapy.
These features might provide a significant improvement in the therapeutic
response to nebulized corticosteroid therapy in uncooperative children who
demonstrate poor compliance to lengthy treatment times with traditional
jet nebulizer systems.