Online Supplemental material Maternal vitamin D status in pregnancy and risk of lower respiratory tract infections, wheezing and asthma in offspring Eva Morales, Isabelle Romieu, Stefano Guerra, Ferrán Ballester, Marisa Rebagliato, Jesús Vioque, Adonina Tardón, Cristina Rodriguez Delhi, Leonor Arranz, Maties Torrent, Mercedes Espada, Mikel Basterrechea, Jordi Sunyer, on behalf of the INMA Project eTable 1. Lower respiratory tract infections, wheeze and asthma prevalences, overall and by area of study, the INMA Project. eTable 2. Association between offspring’s outcomes and selected characteristics, the INMA Project. eTable 3. Association between circulating vitamin D level in pregnancy with selected maternal characteristics, the INMA Project (n=1,724). eTable 4. Meta-analysis of the association* (Odds ratios and 95% confidence intervals) between maternal circulating 25-hydroxyvitamin D (25(OH)D) concentrations in pregnancy and offspring’s respiratory outcomes risk of being diagnosed of LRTI within the INMA Project areas of study. eTable 5. Comparison between included and non-excluded subjects, the INMA Project (N=2,502 children followed until birth). eTable 1. Lower respiratory tract infections, wheeze, and asthma prevalences (n cases/n controls (%)), overall and by area of study, the INMA Project. LRTI at 1 y (n=1,693) Area of study Gipuzkoa Sabadell Valencia Menorca Total 159/260 (38) 163/233 (41) 187/446 (30) 108/137 (44) 617/1076 (37) Wheeze at 1 y (n=1,724) 149/270 (36) 151/242 (38) 167/466 (26) 57/222 (20) 524/1200 (30) Wheezy at 4 y (n=1,233) 83/325 (20) 102/445 (19) 35/243 (13) 220/1013 (18) Asthma at 4-6 y (n=1,233) 91/317 (22) 112/435 (20) 50/228 (18) 253/980 (20) eTable 2. Association between offspring’s outcomes and selected characteristics, the INMA Project. LRTI at 1 year (n=1,693) Sex male; % Firstborn; % Preterm; % Maternal pre-pregnancy body mass index (kg/m2); % <18.5 18.5-25 25-30 >30 Maternal social class; % I-II Managers/Technicians III-IV Skilled manual & nonmanual V Semi-skilled/Unskilled Maternal educational level; % Primary or less Secondary High Maternal asthma; % Maternal smoking pregnancy; % Day care attendance; % Breastfeeding duration (weeks); % 0 >0-16 16-24 >24 Season of birtha; % Winter Spring Summer Yes (n=617) 57 47 5 No p value (n=1,076) 48 <0.001 60 <0.001 4 0.058 Wheeze at 1 year (n=1,724) Yes (n=524) 58 44 5 No p value (n=1,200) 48 <0.001 60 <0.001 4 0.329 Current asthma at 4-6 y (n=1,233) Yes No p value (n=253) (n=980) 56 50 0.077 55 55 0.940 7 4 0.044 4 67 19 9 5 0.114 72 17 7 5 67 19 9 5 0.237 71 17 7 4 64 23 9 4 0.117 71 17 8 36 41 39 0.384 41 35 41 38 0.345 42 30 47 33 0.580 46 23 20 23 20 23 21 32 39 29 9 20 28 0.185 39 32 6 0.011 16 0.034 30 39 31 10 23 31 0.971 39 30 6 0.002 16 <0.001 38 39 24 9 23 35 0.650 40 25 8 0.507 18 0.058 38 28 <0.001 39 28 <0.001 23 28 0.089 14 28 15 43 12 0.250 26 16 46 15 28 17 41 13 0.191 26 16 46 16 29 12 43 13 0.138 27 17 44 23 22 26 30 0.002 23 19 26 21 26 27 0.028 23 20 26 23 22 27 0.943 24 21 Fall a 29 28 27 30 30 Winter: December to February; Spring: March to May; Summer: June to August; Fall: September to November. 29 eTable 3. Association between circulating vitamin D level sin pregnancy with selected maternal characteristics, the INMA Project (n=1,724). Siblings at birth First born No first born Maternal age 20-29 >=30 <20 Maternal ethnic group Caucasian Non Caucasian Maternal BMI pre-pregnancy (kg/m2) <18.5 18.5-25 25-30 >30 p-value* N Mean (SE) 954 770 29.8 11.1 30.8 11.2 0.045 443 1268 13 28.9 10.5 30.8 11.3 25.1 11.9 0.001 1661 55 30.3 11.0 31.0 15.2 0.847 80 1207 310 127 28.1 30.5 29.8 30.6 12.0 11.0 11.5 11.2 0.086 1602 122 30.3 11.2 30.5 10.4 0.801 529 668 527 29.4 10.6 30.0 11.1 31.4 11.7 0.033 610 686 349 31.2 11.5 29.8 10.5 29.7 11.9 0.052 1418 306 30.6 11.2 28.8 10.8 0.013 Maternal asthma No Yes Maternal education level Primary or less Secondary High Maternal social class I-II Managers/Technicians III-IV Skilled manual & non-manual V Semi-skilled/Unskilled Maternal smoking in pregnancy No Yes eTable 4. Meta-analysis of the association* (Odds ratios and 95% confidence intervals) between maternal circulating 25-hydroxyvitamin D (25(OH)D) concentrations in pregnancy and offspring’s respiratory outcomes within the INMA Project areas of study. Area of study Gipuzkoa Sabadell Valencia Menorca P of heterogeneity Combined Fixed effects Random effects * LRTI at 1 y (n=1,693) Wheeze at 1 y (n=1,724) Wheezy at 4 y (n=1,233) Asthma at 4-6 y (n=1,233) 0.56 (0.31-1.03) 0.81 (0.43-1.52) 0.81 (0.48-1.38) 0.48 (0.22-1.06) 0.614 0.71 (0.39-1.30) 1.35 (0.71-2.59) 1.04 (0.61-1.78) 0.64 (0.25-1.63) 0.421 0.99 (0.50-1.97) 0.89 (0.46-1.71) 1.37 (0.49-3.85) 0.786 0.77 (0.39-1.50) 0.95 (0.51-1.76) 1.19 (0.46-3.06) 0.751 0.68 (0.50-0.92) 0.68 (0.50-0.92) 0.94 (0.68-1.30) 0.94 (0.68-1.30) 1.00 (0.65-1.54) 1.00 (0.65-1.54) 0.91 (0.61-1.38) 0.91 (0.61-1.38) Estimates for subjects with circulating 25(OH)D concentrations in the upper cohort- and season-specific quartile in comparison with the lower quartile. All models adjusted for sex, maternal education, siblings at birth, breastfeeding duration, maternal smoking in pregnancy, day-care attendance, maternal asthma, and maternal pre-pregnancy body mass index. eTable 5. Comparison between included and non-excluded subjects, the INMA Project (N=2,502 children followed until birth). Included Not included p value N=1,724 N=878 n=1724 n=691 Child sex; n (%) Male 883 (51) 358 (52) 0.793 n=1724 954 (55) n=1717 70 (4) First born; n (%) Yes Preterm; n (%) Yes Birth weight; grs Mean (sd) n=776 395 (51) 0.040 n=766 41 (5) 0.155 n=1720 n=765 3250.3 (465.1) 3216.9 (526.2) 0.412 n=1724 31.5 (4.3) Maternal age; years Mean (sd) n=1724 Maternal pre-pregnancy BMI (kg/m2); n (%) <18.5 18.5-24.99 25-29.99 ≥ 30 n=686 31.1 (4.5) 0.125 n=759 80 (5) 1207 (70) 310 (18) 127 (7) n=1724 529 (31) 668 (39) 527 (31) 38 (5) 0.195 559 (74) 113 (15) 49 (7) n=757 266 (35) 0.081 281 (37) 210 (28) Maternal social class; n (%) I-II Managers/Technicians III-IV Skilled manual & non-manual V Semi-skilled/Unskilled n=1645 610 (37) 686 (42) 349 (21) n=730 235 (32) 0.059 336 (46) 159 (22) Maternal smoking in pregnancy; n (%) Yes n=1724 306 (18) n=737 0.924 132 (18) Breastfeeding duration (weeks); n (%) 0 >0-16 16-24 >24 n=1724 228 (13) 454 (26) 273 (16) 769 (45) n=589 72 (12) 0.540 158 (27) 107 (18) 252 (43) Day care attendance at age 1y; n (%) Yes n=1724 542 (31) n=536 181 (34) 0.312 Maternal circulating vitamin D levels in pregnancy (ng/mL) Mean (sd) n=1724 30.3 (11) n=534 31.0 (12) 0.315 Maternal education level; n (%) Primary or less Secondary High n=1688 Lower respiratory tract infection at 1 year; n=529 n (%) Yes Wheeze at 1 year; n (%) Yes Wheeze at 4 year; n (%) Yes Current asthma at 4-6 years; n (%) 612 (36) n=1724 524 (30) n=1112 191 (17) n=1111 7 224 (42) 0.012 n=549 160 (29) 0.578 n=391 72 (18) 0.579 n=381 Yes 219 (20) 8 81 (21) 0.515