In order to evaluate the effect of three different primary vaccination intervals on EI vaccine response, 21 unvaccinated thoroughbred foals were randomly divided into three groups of 7 and vaccinated with three different intervals of primary immunization (i.e., with 1, 2 or 3 months intervals between V1 and V2, respectively). The antibody response was measured for up to 1 year after the third immunization V3 (administered 6 months after V2) by single radial hemolysis (SRH) assay. All weanlings had seroconverted and exceeded the clinical protection threshold 2 weeks after V2 and 1 month after V3 until the end of the study. Significant differences were measured at the peak of immunity after V2 and for the duration of the immunity gap between V2 and V3. The group with one month primary vaccination interval had a lower immunity peak after V2 (158.05 ± 6.63 mm2) and a wider immunity gap between V2 and V3 (18 weeks) when compared with other groups (i.e., 174.72 ± 6.86 mm2 and 16 weeks for a two months interval, 221.45 ± 14.48 mm2 and 12 weeks for a 3-month interval). The advantage observed in the group with 1 month primary vaccination interval, which induces an earlier protective immunity, is counterbalance with a lower peak of immunity and a wider immunity gap after V2, when compared with foals vaccinated with 2- and 3-month intervals.