( %FEV 1 ) ( %FEV 1 15%) (P =0.019) (P = 0.005) (airway obstruction);

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1 CHEST Lisa G. Wood, PhD; Heather Powell, MMedSci; Terry Grissell, BSc; Thuy T.D. Nguyen, MD; Darren Shafren, PhD; Michael Hensley, MBBS; and Peter G. Gibson, MBBS 40 ( ) 4 6 wk (n = 26) 15% FEV 1 ( %FEV 1 ) 10 (25%) 30 (75%) ( %FEV 1 15%) (P =0.019) (P = 0.005) (P = ) (airway inflammation); (virus) (airway obstruction); (asthma exacerbation); ACQ = asthma control questionnaire; CCQ = common cold questionnaire; ECP = eosinophilic cationic protein; EV = enterovirus; %FEV 1 = change in FEV 1 percent predicted; ICS = inhaled corticosteroid; IL = interleukin; IQR = interquartile range; MPV = metapneumovirus; PCR = polymerase chain reaction; PEF = peak expiratory flow; RSV = respiratory syncytial virus; RV = rhinovirus 696

2 COPD [1] 81% [2] Pizzichini [10] (IL) -5 (n =6) (n =2) [3 5] FEV 1 ( [2 6 10] 73% 93%) RANTES ( T ) ( 53% 20%) El-Radhi [18] IL-10 IL-5 CD25 [11] (ECP) Corne [12] (RV) 35 d 7d Chang [13] 89% [14] (PEF) [15] RV PEF COPD John Hunter [11] [ ] 25% 5wk PEF [16,17] 7 FEV 1 40% 4 6 wk From the School of Medicine and Public Health (Drs. Wood, Nguyen, Hensley, and Gibson, Ms. Powell, and Mr. Grissell), University of Newcastle, Newcastle, NSW, Australia; and Picornaviral Research Unit (Dr. Shafren), Royal Newcastle Hospital, Newcastle, NSW, Australia. This study was funded by a National Health and Medical Research Council of Australia Project Grant. Correspondence to: Lisa Wood, PhD, Hunter Medical Research Institute, Respiratory and Sleep Medicine, Level 3, HMRI, John Hunter Hospital, Newcastle, NSW 2305, Australia; lisa. wood@newcastle.edu.au / (CCQ) [21] (ACQ) [22] ACQ ( β 2 - ) [22] CHEST

3 [30] FEV 1 ) PCR ( %FEV 1 ) 15% RV EV RSV MPV PCR %FEV 1 15% PCR PCR (TaqMan Applied Biosystems) 12.5% [23] 4wk DNA (RealMasterMix Probe ROX John Hunter kit Eppendorf AG Hamburg Germany) 95 2 min PCR Hunter (HotMaster Taq DNA PCR Eppendorf AG) s 60 1 min (ABI 7500 cycler Applied Biosystems Foster City CA) ( PCR PCR ) (1) ( ) (Buffer RLT Qiagen Hilden Germany) ( PCR PCR ) RNA RNA (PCR) (2) (HEL HEp-2) (3) 1 2 FEV 1 (L) ( 15% 15%) (May- Grnwald ) 400 (Stata version 7 Stata Corporation College [24 25] Station TX) (IQR) (Buffer RLT Qiagen) (%) Kruskal-Wallis (RNeasy kit Qiagen) Fisher χ 2 (P 0.05) RNA Kruskal-Wallis (Kruskal-Wallis 2 (Superscript II RT kit test Stata Corporation) Invitrogen Carlsbad CA) RNA 1 2 DNA Wilcoxon RV (EV) A B (RSV) 40 A B RV 30 (75%) EV 17 (42.5%) [11] (MPV) RNA PCR A 4 (10%) B 1 (2.5%) PCR (Taq-Man RSV 1 (2.5%) probe Applied Biosystems Foster City CA) 3 10 (RV [26] EV [27] RSV A B [28] MPV [29] (25%)

4 1 (1) P (n = 26) (n = 30) (n =10) / 13.0 ( ) 10.9 ( ) 15.6 ( ) (2) (76.9) 24 (88.9) 9 (90) (2) / 7.2 ( ) 7.06 ( ) 14.1 ( ) (3) ICS / µg d -1(3) 650 ( ) 650 ( ) ( ) (4) (3) (1) (2) (3) (4) (IQR) (%) Fisher Kruskal-Wallis P (75%) 1 ACQ (ICS) 1 ( 2A) 2 ( 2B) ( 4) ( 3 1) %FEV 1 FEV 1 FEV 1 CCQ ACQ 2 CCQ 1/4 2 (1) P 10 (33.3) 30 2 (20.0) (2) (2) 4(20.0) 0 4 (20.0) 3 (42.9) 12 (60.0) 4 (57.1) /d 2(1 3) 30 2 (1 4) (3) (1) (2) (3) (IQR) (%) Fisher Wilcoxon CHEST

5 4 6 wk 3 (1) P 1 2 P FEV 1 L 1.34 (2) 2.03 (2) (2) ( ) ( ) ( ) ( ) ( ) % (2) (2) (2) ( ) ( ) ( ) ( ) ( ) CCQ (3) (2) ( ) ( ) ( ) ( ) ( ) (0 2.5) (0 1) (0 6) (0 0) (0 0) (1.5 4) (0 3) (0 7) (0 4.5) (0 2) (0 2) (0 1) (0 3) (0 0) (0 1) (1 4) (0 1) (2 6) (1 2) (0 2) ACQ 2.29 (2) (2) ( ) ( ) ( ) ( ) ( ) (0.5 4) (0 1) (2 5) (0 2) (0 0) (0 3) (0 1) (0 3) (0 2) (0 2) (0 3) (0 2) (1 5) (0 3) (0 0) (1 4) (0 1) (0 4) (0 3) (0 1) 1wk (1 3.5) (0 2) (1 4) (0 4) (0 1) (1 4) (0 2) (3 5) (0 3) (0 2) FEV 1 /% (4 6) (1 2) (2 4) (2 4) (0 2) (1) (2) (IQR) P 0.05 (3) 5 700

6 P < P = NS P < P = P = FEV 1 /L ACQ CCQ 1 P 1 2 NS 25% 5wk Seemungal [16] 2 COPD [17] (FEV 1 ) / 10 4 ml -1 / 10 6 ml -1 0 ICS 2 ( 1) (A) (B) [16] COPD Kruskal-Wallis 2 0 [17] FEV 1 ( 2) 3 CCQ 6 A 4 6 wk P = (CCQ )( 1) [31] ACQ 2 2 (ACQ 0.75) (ACQ 1.5) [31] 300 B P = (FEV 1 ) 200 %FEV 1 ( 1 3) CHEST

7 4 (1) P 1 2 P / 10 6 ml (2) ( ) ( ) ( ) ( ) ( ) / 10 4 ml (2) ( ) ( ) ( ) ( ) ( ) / 10 4 ml ( ) ( ) ( ) ( ) ( ) / 10 4 ml ( ) ( ) ( ) ( ) ( ) / 10 4 ml ( ) ( ) ( ) ( ) ( ) / 10 4 ml ( ) ( ) ( ) ( ) ( ) (1) (2) (IQR) P 0.05 [37] ( 2) ( 4) IL-8 ECP [38] [11] [39] [40] ( 2) [41 42] ( 4) ICS ( 1) ICS 3 2 [32] -α IL-1 [33] IL wk 1/4 ECP [34] C4 [35 36] M 2 (25%) 702

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