J ALLERGY CLIN IMMUNOL

VOLUME 127, NUMBER 2

Abstracts AB87

SUNDAY

323 Effect of Budesonide/Formoterol Pressurized Metered-DoseInhaler (BUD/FM pMDI) on Proportion of the Mean PeakForced Expiratory Volume in 1 Second (FEV1) Over Time inPatients With Chronic Obstructive Pulmonary Disease orPersistent Asthma

F. Ahmad, J. G. Zangrilli, U. J. Martin; AstraZeneca LP, Wilmington, DE.

RATIONALE: The spirometric response to an inhaled medication over

time provides insight into the peak effects characteristic of bronchodila-

tors. We assessed lung function as a proportion of maximal FEV1 over

time (ie, greatest change from baseline or peak FEV1) to evaluate BUD/

FM pMDI response at the beginning of the dosing interval.

METHODS: Analysis of 12-hour serial spirometry data (day of random-

ization and 2 subsequent visits) from 2 double-blind studies of patients >_40

years with moderate to severe chronic obstructive pulmonary disease

(COPD; I [12-month]: NCT00206167 [Drugs. 2008;69:549]) or >_12 years

with moderate to severe persistent asthma (II [12-week]: NCT00652002

[Drugs. 2006;66:2235]) who were randomized to twice-daily BUD/FM

pMDI 320/9 mg was conducted. The mean percentage change from base-

line FEV1 at each spirometry time point as a proportion of peak FEV1

(eg, change from baseline FEV1 peak) was assessed.

RESULTS: Study I (COPD, n5121): The peak percentage change from

baseline FEV1 occurred at 2 hours postdose for all 3 visits and ranged

from 27-29% across visits. Clinically significant bronchodilation (>_15%

change from baseline in predose FEV1) occurred within 5 minutes, repre-

senting 64-81% of mean peak FEV1. Study II (asthma, n5124): The peak

percentage change from baseline FEV1 occurred 1-3 hours postdose and

ranged from 19-21% across visits. Clinically significant bronchodilation

was achieved within 15minutes, representing 78-87% of mean peak FEV1.

CONCLUSIONS: Patients with COPD and asthma achieved a high pro-

portion of peak FEV1 rapidly (within 5 and 15 minutes, respectively), as

measured after the morning dose of BUD/FM pMDI 320/9 mg.

324 Inhibitory Effect of Constituents Isolated from Ganodermalucidum on LPS-stimulated Macrophages TNF-a Production

Y. Chuang, C. Liu, N. Yang, Y. Song, B. Liang, X. Li; Pediatric Allergy &

Immunology, The Mount Sinai School of Medicine, New York, NY.

RATIONALE: TNF-a is a pro-inflammatory cytokine that had been im-

plicated in many aspects of asthmatic airway pathophysiology. ASHMI

(Antiasthma Simplified Herbal Medicine Intervention) was shown to

have therapeutic effects in an animal model of asthma and in a controlled

clinical trial. Ganoderma lucidum (Ling-Zhi) one of the three herbs in

ASHMI, has an inhibitory effect on TNF-a production similar that of

ASHMI. We performed activity-guide isolation to identify the bio-active

compounds in Ling-Zhi that inhibit TNF-a production.

METHODS: Liquid-liquid extraction, silica gel column chromatography,

sephadex LH-20 column chromatography and preparative HPLC were

used to obtain pure compounds from Ling-Zhi. The effect of these isolated

compounds on TNF-a production by RAW 264.7 macrophages was exam-

ined. Cell viability was measured by MTT assay.

RESULTS: 16 pure compounds (1-16) were isolated from Ling-Zhi.

Inhibitory activity of these compounds on macrophages TNF-a production

was tested at different doses. Comp.8 and Comp.11 significantly inhibit

TNF-a production at 40mg/mL (48.4% and 47.7%, respectively, P

<0.0001 vs untreated cells). A synergistic effect was obtained when

Comp.8 (20mg/mL) and Comp.11 (20mg/mL) were combined (55.7% inhi-

bition). No compounds were cytotoxic at concentrations up to 40mg/mL.

CONCLUSIONS: Individual constituents (Comp.8, Comp.11) and the

combination (Comp.8 and Comp.11) isolated from Ling-Zhi inhibit

TNF-a production in vitro and may have potential for treating allergic in-

flammatory conditions.

325 Successful Treatment of Acute HemorrhagicLeukoencephalitis, a Novel Phenotype for ComplementFactor I Deficiency

L. Broderick, C. Gandhi, J. L. Mueller, C. D. Putnam, K. Shayan, K. S.

Peterson, S. S. Aceves, R. M. Sheets, B. M. Peterson, R. O. Newbury,

H. M. Hoffman, J. F. Bastian; University of California, San Diego, San

Diego, CA.

RATIONALE: Acute Hemorrhagic Leukoencephalitis (AHLE) is a rare,

acute onset demyelination disorder, causing rapid neurologic deterioration

and mortality. It is most often described as a post-infectious complication

of an upper respiratory illness, although its precise pathophysiology re-

mains unclear. We describe two pediatric patients with AHLE with com-

plement factor I (CFI) deficiency whose successful treatment included

the interleukin-1 (IL-1) receptor antagonist, anakinra, implicating a role

for CFI and IL-1 in demyelinating neurodegenerative disorders.

METHODS: Extensive clinical evaluation of two patients presenting with

AHLE revealed complement abnormalities, specifically related to the alter-

native pathway and its regulator, CFI. Gene sequencing of the CFI coding

regions of the patients and their immediate families was performed on

peripheral blood samples. Immunohistochemical analysis of fixed brain

biopsy tissue was used to investigate the complement pathway and the

mechanism behind the successful use of anakinra.

RESULTS: Both patients received aggressive management with steroids

and immunoglobulin. Treatment with anakinra ultimately led to improve-

ment of clinical status and return to neurologic baseline in both patients.

Two novel mutations in CFI were identified in our patients, which were

not identified in controls. Additionally, immunohistochemical evaluation

of brain biopsy tissue demonstrates increased C3, membrane attack com-

plex (MAC) and IL-1 positivity compared to normal control tissue.

CONCLUSIONS:AHLE is a novel phenotype for CFI deficiency. The up-

regulation of C3 andMACwith subsequent demyelination observed in our

patients supports a role for uncontrolled complement in AHLE, and sug-

gest modulation of innate immunity by anakinra as an important therapy

for neurodegenerative disorders on the demyelination disease spectrum.

326 Complement C6 Null Mutations in Extreme PrematurityT. P. Atkinson1, N. Ambalavanan1, R. A. Kaslow1, M. Peralta-

Carcelen1, S. S. Cosby1, V. A. Phillips1, C. Li2, Y. Dai1; 1University of

Alabama at Birmingham, Birmingham, AL, 2State Perinatal Program,

Alabama Dept of Public Health, Montgomery, AL.

RATIONALE: Since the prevalence of Complement C6 null alleles is in-

creased among African-Americans and the incidence of premature birth is

likewise higher among African-American mothers, we conducted a study

to assess a possible association of complement C6 null alleles in

African-American mothers of extremely low birth weight infants

(ELBW) and ELBWAfrican-American infants.

METHODS:We screened 78 ELBW infants and 55 mothers of ELBW in-

fants for three loss-of-function C6 mutations using PCR and heteroduplex

analysis.

RESULTS: Three infants (3.9%) were found to be heterozygous for a C6

null mutation, a prevalence similar to that in our historical control group

(4.9%). Five mothers were heterozygous for a C6 null allele, and one

was a compound heterozygote indicating complete deficiency; this preva-

lence was significantly higher than in our historical control group (p 50.029). Interestingly, six sets of twins and one set of triplets were born to

cohort mothers, and three of the mothers who had given birth to twins

were heterozygous for C6 null alleles (p5 0.022). Five of the six mothers

bearing C6 null alleles compared with 11 of the other 29 evaluable mothers

had histological chorioamnionitis (p 5 0.073).

CONCLUSIONS: These data suggest that partial C6 deficiency and, by

implication, partial deficiency of other terminal complement components

represent a risk factor for extreme prematurity, and part of this risk may

be related to an increased prevalence in multiple births in mothers bearing

C6 null alleles.