Hamstring motor behaviour and resultant antagonist torque during knee extension has been quantified in uninjured individuals however, the effect of ACL rupture and ACL reconstruction (ACLR) on the morphology of hamstring antagonist torque generated in vivo is unknown. The purpose of this cross-sectional study was to quantify the hamstring antagonist torque generated in vivo during isokinetic knee extension in ACLD and ACLR patients relative to uninjured control subjects. Ten male ACL deficient (ACLD) subjects (18-35 years), 14 matched males who had undergone ACLR using the bone-patellar tendon-bone graft and 22 matched male control subjects participated. We used a mathematical model to estimate the opposing torque generated by the hamstrings during isokinetic knee extension in 10 degrees intervals from 80 degrees to 10 degrees knee flexion. Control group hamstring antagonist torque was significantly lower at 80-70 degrees knee flexion compared with that of the ACLD (% Diff=40.2; p=0.019) and ACLR (% Diff=34.8; p=0.036) groups. For all subject groups, hamstring antagonist torque demonstrated a descending-ascending curve; decreasing significantly from 80-70 degrees to 50-40 degrees knee flexion (% Diff=40.8 to 63.3; p=<0.001 to 0.009) but then increasing significantly from 50-40 degrees to 20-10 degrees knee flexion (% Diff=37.6 to 59.0; p=<0.001 to 0.012). ACL status and therefore, the ACL-hamstring reflex has little effect on the magnitude of hamstring antagonist torque generated during quadriceps-induced knee joint loading. Capsular afferents are thought to dictate the hamstring torque profile which decreased then increased during knee extension to maintain dynamic joint stability.