This study examined the status of cortisol and C-reactive protein (hs-CRP) as stress and inflammatory markers in airplane pilots. The study comprised 78 airplane pilots and 54 healthy control subjects who were recruited from North Marka Laboratory, Amman, Jordan. Anthropometric data including age, weight, height, body mass index (BMI), and blood pressure were determined using standard procedures. Biochemical parameters, including lipid profile, liver and kidney function tests, cortisol, and hs-CRP levels, were measured using conventional laboratory techniques. Results from this study showed that serum levels of cortisol and hs-CRP were higher in pilots than in controls (19.1 ± 7.1 versus 12.7 ± 9.7 µg/dL, p < 0.05; and 4.2 ± 5.3 versus 2.3 ± 1.6 mg/L, p = 0.015, respectively). The levels of cortisol and hs-CRP were not different among pilots when compared according to flying hours. Based on BMI however, the levels of hs-CRP were significantly higher in pilots with BMI > 25 kg/m2; p < 0.050. Bivariate correlational analyses revealed positively significant correlation between hs-CRP and cortisol levels in pilots as well as in all subjects combined (r = 0.250, p = 0.027 versus r = 0.294, p = 0.029); respectively. In conclusion, this study demonstrate that pilots had increased serum levels of cortisol and hs-CRP that may highlights the potential impact of occupational stress on physiological health.