BACKGROUND: Persistent headward fluid shift and mechanical unloading cause neuro-ocular, cardiovascular, and musculoskeletal deconditioning during long-term spaceflight. Lower body negative pressure (LBNP) reintroduces footward fluid shift and mechanical loading.METHODS: We designed, built, and tested a wearable, mobile, and flexible LBNP device (GravitySuit) consisting of pressurized trousers with built-in shoes to support ground reaction forces (GRF) and a thoracic vest to distribute load to the entire axial length of the body. In eight healthy subjects we recorded GRF under the feet and over the shoulders (Tekscan) while assessing cardiovascular response (Nexfin) and footward fluid shift from internal jugular venous cross-sectional area (IJVa) using ultrasound (Terason).RESULTS: Relative to normal bodyweight (BW) when standing upright, increments of 10 mmHg LBNP from 0 to 40 mmHg while supine induced axial loading corresponding to 0%, 13 ± 3%, 41 ± 5%, 75 ± 11%, and 125 ± 22% BW, respectively. Furthermore, LBNP reduced IJVa from 1.12 ± 0.3 cm² to 0.67 ± 0.2, 0.50 ± 0.1, 0.35 ± 0.1, and 0.31 ± 0.1 cm², respectively. LBNP of 30 and 40 mmHg reduced cardiac stroke volume and increased heart rate while cardiac output and mean arterial pressure were unaffected. During 2 h of supine rest at 20 mmHg LBNP, temperature and humidity inside the suit were unchanged (23 ± 1°C; 47 ± 3%, respectively).DISCUSSION: The flexible GravitySuit at 20 mmHg LBNP comfortably induced mechanical loading and desired fluid displacement while maintaining the mobility of hips and knee joints. The GravitySuit may provide a feasible method to apply low-level, long-term LBNP without interfering with daily activity during spaceflight to provide an integrative countermeasure.Petersen LG, Hargens A, Bird EM, Ashari N, Saalfeld J, Petersen JCG. Mobile lower body negative pressure suit as an integrative countermeasure for spaceflight. Aerosp Med Hum Perform. 2019; 90(12):993-999.